فهرست مطالب

Pharmaceutical Research - Volume:18 Issue:3, 2019
  • Volume:18 Issue:3, 2019
  • تاریخ انتشار: 1398/04/10
  • تعداد عناوین: 45
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  • Theaj Prakash Upputuri Ravi, Abul Kalam Azad Mandal* Pages 1137-1146
    Drug release kinetics plays an important role in determining the mechanism of drug release, which in turn helps in formulating controlled/sustained release formulations.In our study, different concentrations of green tea polyphenols (GTP) were encapsulated into casein nanoparticles which showed a maximum encapsulation efficiency (76.9%) at a GTP concentration of 5 mg/mL. The casein nanoparticles were characterized through particle size analysis, zeta potential, AFM, and HR SEM, followed by molecular docking studies, which confirmed the binding of GTP to casein nanoparticles. In-vitro release studies carried out at different temperatures and pH showed no significant difference in the release pattern, but the release was prolonged even up to 48 h. On varying pH of the release medium, an increase in the percentage of release was observed as the pH shifted from acidic to basic. All release data showed good correlation with Zero order kinetics, an ideal model for release of drugs from nanoparticulate sustained release formulations, with anomalous mode of drug transport. Antioxidant activity of the released GTP determined through DPPH assay showed potent antioxidant effect of GTP even after 48 h of its release.Our data indicated that casein nanoparticles could be used as a potent vehicle for the delivery of GTP for achieving a sustained release.
    Keywords: Anomalous transport, Casein nanoparticles, Green tea polyphenols, Mathematical modeling, sustained release, Zero order kinetics
  • Meizhen Li, Wei Qiang, Zhou Wen, Linling Li, Lei Wang*, Zeneng Cheng* Pages 1147-1155
    Complex pharmacokinetic (PK) properties including nonlinear elimination were encountered by some monoclonal antibodies (mAbs), and classic compartment models sometimes failed to appropriately describe those properties. In this work, a new model was built on a comprehensive analysis of the complex elimination of mAbs. This new model was firstly utilized to fit with the single-dose plasma concentration data of bevacizumab in beagle dogs receiving an intravenous administration of 2.5 mg/kg bevacizumab. Then, the optimal PK parameters from fitting with the single-dose PK data were employed into the multiple-dose mathematical expressions to predict bevacizumab’s multiple-dose PK profiles. One-compartment model recommended as the optimal classic model by DAS 2.0 software was set as a control. As a result, new model fitted better with the single-dose PK profiles of bevacizumab with smaller weighted residual sum of squares and higher fitting degree compared with the classic model. Importantly, new model also accurately predicted the multiple-dose PK profiles of bevacizumab and performed well at the single-to-multiple transition. In conclusion, the new model reasonably explained the complex elimination of bevacizumab, and it might play a big role in the PK studies of bevacizumab and other mAbs.
    Keywords: Elimination, Metabolism, Modeling, Monoclonal antibodies, Simulations
  • Keshvad Hedayatyanfard, Shadab Bagheri, Khoulenjani, Ali Hashemi, Seyed Ali Ziai* Pages 1156-1167
    The antimicrobial activity of a wound dressing is a key factor for preventing and treating wound infection. The current study evaluated the physiochemical properties and antimicrobial activities of semi-IPNs (interpenetrating polymer networks) based on chitosan/polyvinyl alcohol (PVA) films and nanofibers as candidates for wound dressings and investigated the effects of morphologies (nanofibrous mats and films), crosslinking conditions of chitosan chains (uncrosslinked and crosslinked with genipin), and the presence of antibacterial drug (doxycycline) on their physicochemical and antibacterial properties. The morphology, chemical structure, fluid uptake, water vapor transmission rate, antimicrobial activity, and doxycycline release profile were assayed using SEM, FTIR spectroscopy, swelling test, permeation test, agar diffusion antibiogram, and dissolution test, respectively. The results demonstrated that crosslinking chitosan with genipin reduced the diameter of nanofibers, fluid uptake, and drug release from both nanofiber mats and film samples. According to the results, wound dressings with film morphology have better antimicrobial activity than those with nanofiber. The chitosan/PVA/Doxycycline 1% film has the potential for use as an antimicrobial wound dressing.
    Keywords: PVA, Chitosan, doxycycline, Film, Nanofiber, Antimicrobial, Genipin
  • Fereydoon Abedi, Gaballu, Soheil Abbaspour, Ravasjani, Behzad Mansoori, Reza Yekta, Hamed Hamishehkar, Ali Mohammadi, Gholamreza Dehghan, Behrooz Shokouhi, Shaho Ghahremani Dehbokri*, Behzad Baradaran* Pages 1168-1179
    Erlotinib (ELT) as a small molecule with poor solubility, poor bioavailability, and instability in gastrointestinal environment, has been considered as a therapeutic agent for Non-Small-Cell Lung Cancer (NSCLC) therapy through oral administration. In the present study, ELT-liposome and ELT-NLCs were successfully prepared and characterized by assessment of the particle size, zeta potential (ZP), polydispersity index (PDI), encapsulation efficiency (EE) and drug loading (DL). DAPI staining and Flow cytometry techniques were employed to probe anticancer activities of the optimal formulations. The obtained results indicated that the average size of optimized ELT-NLCs was 109±2 nm, while the optimal formulation of ELT-liposome was 130±4 nm. In addition, the values of EE, DL and cellular uptake were higher in ELT-NLCs than ELT-liposome. Moreover, the stability of ELT-NLCs and ELT-liposome were not significantly changed (P > 0.05) within storage time. The results of anti-cancer assessment showed that ELT-NLCs caused more cell viability reduction than ELT-liposome and free ELT. According to the Flow cytometry and DAPI staining results, the exposed A549 cells with ELT-NLCs had more rate of apoptosis than ELT-liposome. The obtained data from this study clearly showed that ELT-NLCs had better anti-cancer activity than ELT-liposome, which may be is related to the effective nano particle size, PDI, EE, and DL of ELT-NLCs.
    Keywords: Erlotinib, Liposome, NLCs, A549 cells, Lung Cancer
  • Nasim Khorshidian, Arash Mahboubi, Naser Kalantari, Hedayat Hosseini, Mojtaba Yousefi, Masoumeh Arab, Adriano Gomez Da Cruz, Amir Mortazavian*, Fatemeh Sadat Mahdavi Pages 1180-1195
    Many herbs and spices have been recommended traditionally as galactagogues and several commercial formulations prepared using herbs. Due to the presence of various compounds such as polyphenols, flavonoids, isoflavones and terpenes, bitter and stringent taste is elicited that make the consumption of these herbal preparations unpleasant. Moreover, these compounds are unstable when exposed to environmental conditions. In this regard, different approaches are used for taste masking such as microencapsulation. In the present study, microcapsules containing galactagogue herbs extract were developed through emulsification/external gelation and Box-Behnken design was used to investigate the effects of independent variables (sodium alginate: 1-1.5%, calcium chloride: 0.2-1% and extract concentrations: 1-5%) on encapsulation efficiency (EE%). Following evaluation of the model, the optimum condition of encapsulation process was selected as 1.49% sodium alginate, 0.84 CaCl2 and 1.58% extract with EE% of 77.97%. Microcapsules had an acceptable spherical morphology and the results of Fourier transform-infrared spectroscopy (FTIR) revealed the presence of the extract within the microcapsules. The mean diameters of the uncoated and chitosan-coated microcapsules were 52 and 123 μm and encapsulation yield was 50.21 and 69.7%, respectively. The polydispersity index of 0.45 and 0.48 were an indicative of polydisperse nature of the microcapsules. The loss of flavonoids in microcapsules stored at two different temperatures was insignificant. The in vitro release in simulated gastric fluid (SGF; pH 1.2) and simulated intestinal fluid (SIF; pH 7.4) were 48.1% and 80.11%, respectively during 24 h. The prepared extract-loaded microcapsules have potential to be used in matrices with neutral pH.
    Keywords: Galactagogues, Herbal extract, Microencapsulation, Sodium alginate, Chitosan, External gelation
  • Zahra Namazi, Teherh Sadat Jafarzadeh Kashi, Mohammad Erfan*, Farhood Najafi, Leila Bakhtiari, Seyed Roholah Ghodsi, Hassan Farhadnejad Pages 1196-1211
    The present study deals with the fabrication of ibuprofen-mesoporous hydroxyapatite (IBU-MHA) particles via the incorporation of ibuprofen (IBU)—as a nonsteroidal anti-inflammatory drug—into mesoporous hydroxyapatite nanoparticles (MHANPs) using an impregnation process, as a novel drug delivery device. MHANPs were synthesized by a self-assembly process using cetyltrimethylammonium bromide (CTAB) as a cationic surfactant and 1-dodecanethiol as a pore expander under basic condition. The focus of the present study was to optimize the incorporation of IBU molecules into MHANPs under different loading conditions. The synthesized MHANPs and IBU-MHA particles were confirmed by X-ray diffraction (XRD), fourier-transform infrared spectroscopy (FTIR), brunauer–emmett–teller (BET), transmission electron microscopy (TEM), and thermal analysis (TGA). Drug loading (DL) efficiency of IBU-MHA particles was determined by ultraviolet–visible (UV-Vis) spectroscopy, and indicated that the optimized IBU-MHA particles with high DL (34.5%) can be obtained at an IBU/ MHANPs ratio of 35/50 (mg/mg), impregnation period of 24 h, and temperature of 40 °C using ethanol as solvent. In-vitro drug release test was carried out to prove the efficiency of IBU-MHA particles as a sustained drug delivery system. A more sustained and controlled drug release was observed for this particles, indicating that it may be have good potential as drug reservoirs for local drug release.
    Keywords: Mesoporous, Nanoparticle, Hydroxyapatite, Ibuprofen, Drug delivery
  • Farhad Ahmadi*, Nilofar Rahmani Pages 1212-1223
    An efficient and quantitative two phase freezing (TPF) method coupled with high performance liquid chromatography and UV-Vis detector was developed for the extraction, clean up and determination of clomiphene citrate (CLC) in plasma samples. The separation of two miscible solvents by TPF method permits that the CLC was efficiently removed from proteins and transferred into the relative aprotic dipolar organic phase and in consequence, gave a higher recovery. The TPF method was compared to conventional liquid-liquid extraction and it gave more clean solution with better reproducibility. Linear range, limit of detection and limit of quantification for CLC in plasma were obtained in the range of 0.06-18, 0.02 and 0.06 µg ml-1, respectively. The intraday and interday reproducibility for concentration of 1.0 µg ml-1 (%RSD) were 3.2% and 4.6%, respectively. In addition, the trueness, ruggedness, and realistic of TPF were assessment. Finally, several real plasma samples were successfully analyzed using the developed method.
    Keywords: Clomiphene citrate, two phase freezing method, Liquid-liquid extraction, Doping control, plasma extraction
  • Mitra Bayat, Mohammad Saber Tehrani*, Farzad Kobarfard, Seyed Waqif Husain, Hassan Yazdanpanah Pages 1224-1238
    Persistent organic pollutants, (POPs), are vast distributed compounds in environment which are recognized as one of the global pollution problems. These groups of materials being dangerous due to their high stability are accumulated in animal tissues and occurring in the food chain. One of the major paths through which persistent organic combinations access to human body is consuming polluted foods, particularly, fishes. Among aquatic animals, trout as one of the mostly consumed fishes in Tehran’s food basket was studied. In this study two categories of persistent organic pollutants: Organochlorine pesticides (OCPs) including HCB, Dieldrin, Methoxychlor, α-, ϒ-Chlordane, α-, β-Endosulfan and o,p’-DDE, p,p’-DDE, o,p’-DDT, p,p’-DDT and the second group Polychlonitated biphenyls (PCBs) including seven PCB congeners which are called indicator PCBs (IUPAC nos.: 28,52,101,118,138,153 and 180) were determined in trout by GC-MS/MS in MRM monitoring mode and LLE extraction. The average recoveries of OCPs and PCBs at five concentration levels were in the range of 73-112%. The relative standard deviations of POPs in fish were in the range of 1.4-17.9% for all of the concentration levels. Limit of detections (LODs) and limit of quantitations (LOQs) were between 0.6-8.3 and 2-25 µg/kg, respectively. The results indicated the presence of organochlorine pesticides in trout and the levels of p,p’-DDE and p,p’-DDT were within the range of < LOQ -12.83 and < LOQ -10.2 ng/g ww (wet weight), respectively. According to the results, OCPs residues were lower than maximum residue levels set by European Council Directives.
    Keywords: Persistent Organic Pollutant (POPs), Polychlonitated biphenyls, Organochlorine pesticides, Trout, Liquid Liquid Extraction, GC-MS, MS, Iran
  • Zohreh Doroudi, Ali Niazi* Pages 1239-1252
    Quantitative structure-activity relationship (QSAR) analysis has been carried out with a series of 107 anti-HIV HEPT compounds with antiviral activity, which was performed by chemometrics methods. Bi-dimensional images were used to calculate some pixels and multivariate image analysis was applied to QSAR modelling of the anti-HIV potential of HEPT analogues by means of multivariate calibration, such as principal component regression (PCR) and partial least squares (PLS). In this paper, we investigated the effect of pixel selection by application of genetic algorithms (GAs) for the PLS model. GAs is very useful in the variable selection in modelling and calibration because of the strong effect of the relationship between presence/absence of variables in a calibration model and the prediction ability of the model itself. The subset of pixels, which resulted in the low prediction error, was selected by genetic algorithms. The resulted GA-PLS model had a high statistical quality (RMSEP = 0.0423 and R2 = 0.9412) in comparison with PCR (RMSEP = 0.4559, R2 = 0.7929) and PLS (RMSEP = 0.3275 and R2 = 0.0.8427) for predicting the activity of the compounds. Because of high correlation between values of predicted and experimental activities, MIA-QSAR proved to be a highly predictive approach.
    Keywords: Multivariate image analysis, Genetic algorithms, Partial least square, Principal Component Regression, Variable selection, 1-[2-hydroxyethoxy)methyl]-6-(phenylthio)thymine
  • Kamyar Faghihi, Mahdieh Safakish, Tannaz Zebardast, Zahra Hajimahdi*, Afshin Zarghi* Pages 1253-1263
    A series of 2-benzoxazolinone, diazocoumarin and quinazoline derivatives have been shown to inhibit HIV replication in cell culture. To understand the pharmacophore properties of selected molecules and design new anti-HIV agents, quantitative structure–activity relationship (QSAR) study was developed using a descriptor selection approach based on the stepwise method. Multiple linear regression method was applied to relate the anti-HIV activities of dataset molecules to the selected descriptors. Obtained QSAR model was statistically significant with correlation coefficient R2 of 0.84 and leave one out coefficient Q2 of 0.73. The model was validated by test set molecules giving satisfactory prediction value (R2test) of 0.79. Molecules also were docked on HIV integrase enzyme and showed important interactions with the key residues in enzyme active site. These data might be helpful for design and discovery of novel anti-HIV compounds.
    Keywords: Anti-HIV, docking, QSAR, Multiple linear regressions, Stepwise
  • Maryam Mazaheri Tehrani, Mostafa Erfani*, Nour Amirmozafari, Taher Nejadsattari Pages 1264-1276
    Microcin J25 (MccJ25) is a small ribosomally synthesized antimicrobial peptide that is produced by Enterobacteriacea family especially E.coli. The present study focuses on preparation and evaluation of in vitro antimicrobial and biological properties of a new peptide derived from MccJ25. We prepared a MccJ25-derived peptide containing 14 amino acids and a single intra-molecular disulfide bond according to solid phase synthesis strategy. The purified peptide was characterized by Liquid chromatography-mass spectrometry (LC-MS) and Fourier Transform Infrared (FTIR) spectroscopy. 96-well microdilution plate assay was exerted for determination of minimum inhibitory concentration (MIC) of peptide against different bacterial strains. Cytotoxicity of the peptide derivative on HT-29 cell line assayed using MTT test. The final peptide successfully was prepared with purity more than 99.8% as determined by analytical HPLC. The evaluation of antibacterial activity of the peptide against Gram-positive and Gram- negative bacteria revealed that the peptide was very effective against E.coli 35218 with minimum inhibitory concentration (MIC) at dose 3.9 µM. The hemolytic activity toward human erythrocytes was very minimal below 0.3 %. The cell viability percentage of HT-29 cell line after 24 hours of contact with the peptide was more than 83%. The high sensitivity of E.coli strain to this new peptide derived from MccJ25 and through minimal toxicity to cancerous cell, suggesting that above synthesized peptide could be considered as a bioactive compound for further investigations.
    Keywords: Microcin J25 derivative, Synthesis, Biological activity, E.coli, Cytotoxicity
  • Saeed Balalaie*, Morteza Malakoutikhah, Meritxell Teixido, Vaezeh Fathi Vavsari, Ernest Giralt, Yaghoub Haghighatnia, Fatima Hamdan, Armin Arabanian Pages 1277-1287
    An efficient approach for the synthesis of norbuprenorphin derivatives through coupling of enkephalins and norbuprenorphine intermediate is described. Norbuprenorphine derivative was synthesized from thebaine and then, its reaction with succinic acid and phthalic acid was also studied. Meanwhile, the synthesis of enkephalins was done using solid phase peptide synthesis approach. Furthermore, after cleavage of the peptide from the surface of the resin, the coupling of enkephalins with norbuprenorphine derivative was done using TBTU as a coupling reagent then the derivatives were purified using preparative high-pressure liquid chromatography and their structures were confirmed using high-resolution mass spectrometry data. Later, their permeability across membranes was investigated. After PAMPA studies, it was found that the permeability of all norbuprenorphin-enkephalin derivatives was increased, however, succinic and phthalic acid derivatives showed higher permeability than norbuprenorphine-Leu-enkephalin.
    Keywords: Enkephalin, Permeability, norbuprenorphine, membrane, coupling, Solid phase peptide synthesis
  • Mehdi Valipour, Issa Davaji, Niusha Abedi, Mahsa Rajabi, Tuba Tüylü Küçükkılınç, Beyza Ayazgök, Hamid Irannejad* Pages 1288-1298
    Alzheimer’s disease (AD) is a neuroinflammatory based pathologic state in which β-amyloid aggregates are a major devastating agents. In this study, a series of 2-hydroxyiminoethanones were synthesized and evaluated as anti-inflammatory in carrageenan and formalin tests and inhibitors of β-amyloid aggregation. Compounds 1-10b were synthesized through a two-step reaction.
    Results
    Compounds 1-5b showed more β-amyloid disaggregation ability than reference drugs rifampicin and donepezil and compound 2b was the best compound in this series and could reduce the extent of amyloid aggregation to 50.9%. Interestingly, compounds 1b and 3b showed significant anti-inflammatory activity in carrageenan-induced paw edema compared to control group and equivalent to the reference drug indomethacin. 2-Hydeoxyiminoethanones are privileged scaffold for further drug research and development as anti-neuroinflammatory and neuroprotective agents.
    Keywords: Synthesis, Anti- inflammatory, Alzheimer's disease, hydroxyiminoethanone, β-amyloid aggregation, Cyclooxygenase
  • Maryam Baharloui, Sayed Ahmmad Mirshokraee, Azam Monfared, Mohammad Hassan Houshdar Tehrani* Pages 1299-1308
    Cancer disease is a great concern in the worldwide public health and current treatments do not give satisfactory results, so, developing novel therapeutic agents to combat cancer is highly demanded. Nowadays, anticancer peptides (ACPs) are becoming promising anticancer drug candidates. This is due to several advantages inherited in peptide molecules, such as being usually with small size, high activity, low immunogenicity, good biocompatibility, diversity of sequence, and more modification sites for functionalization. To get benefit of these merits, in this work, we synthesized a new series of triazole- based analogues with peptide scaffold by employing click chemistry and evaluated their anticancer activities against breast, colon cancer cell lines as well as fibroblast cells using MTT assay. Our results suggest that peptide scaffolds containing 1H-1, 2, 3-triazole ring group are toxic against colon and breast cancer cells viability, and this effect was more pronounced on MDA-MB-231 cells compared with MCF-7 breast cells. As a conclusion, these designed peptide analogues may be good and safe candidates as future anticancer agents.
    Keywords: Click chemistry, Triazole rings, Peptide analogues, Cancer, MTT assay
  • Osama Ahmed Abbas*, Issa Ghada Ibrahim, Abdel, Gawad Eman Ismail Pages 1309-1322
    With the advance in nanomedicine, the present study was conducted to explore the possible therapeutic role of intravenous nano- hydroxyapatite (nano-HAp) into male rats after chronic exposure to aluminum chloride (AlCl3). This exposure interposed DNA fragmentation, apoptosis, alters oxidant/antioxidant status as well as change in content of neurotransmitters. The rats were injected with 100 mg/kg. body weight (b.w.) of AlCl3 intraperitoneally for 90 days, after then nano-HAp was injected intravenously (i.v.) three times per week at a dose level 100 mg/kg b.w.Based on the results obtained, it can be concluded that the treatment with the prepared nano-HAp restrains the damage inflicted on brain modulating by lipid oxidation products and decreased the susceptibility of apoptotic cells death with subsequent repaired the fragmented DNA as well as improved the synthesis of neurotransmitters. The most salient findings for nano-HAp treatment are that most of the pathological changes due to AlCl3 administration were disappeared.
    Keywords: nano- hydroxyapatite, Aluminum chloride, nuclear respiratory factor1, fragmented DNA, caspase3
  • Ali Jahanian, Najafabadi, Mina Mirian, Fatemeh Rouhani, Kazem Karami, Mahboubeh Hosseini Kharat, Hojjat Sadeghi, Aliabadi* Pages 1323-1331
    Today, development of resistance to anticancer drugs (including cisplatin) is noticed as a major problem. Recently several studies demonstrated that palladium complexes showed remarkable cytotoxic effects against K562 cell line and could be used efficiently for treatment of many human cancers including leukemia. Hereof, K562 cells were made resistant to cisplatin using increasing concentration of cisplatin up to 4.5, and then cytotoxic effect of synthesized palladium complex was evaluated on this sub-line using MTT assay. Annexin V/PI staining using flow cytometry and scanning electron microscopy (SEM) were performed to find out the mechanism of the observed cytotoxicity. Results indicated that tested compounds had a noticeable cytotoxic effect on K562 cells 80 times more than cisplatin. Palladium complex also showed significant cytotoxicity on resistant K562 sub-line. Flow cytometry and SEM results revealed that these compounds exert their cytotoxic effect via apoptosis and it could be concluded that the novel synthesized palladium complex might be a good candidate for replacing cisplatin in case of treatment of cisplatin resistant tumors.
    Keywords: K562 cells, Cisplatin Resistance, Palladacyclic complex, MTT assay, Apoptosis assay
  • Hamid Khaneghahi Abyaneh, Alireza Bahonar*, Negin Noori, Hassan Yazdanpanah, Mohammad Hossein Shojaee Aliabadi Pages 1332-1340
    Milk would be contaminated with Aflatoxin M1 (AFM1) if it was obtained from lactating animal which fed with feedstuffs containing Aflatoxin B1 (AFB1). AFM1 is classified as group 2B, possibly carcinogenic to humans and exposure to AFM1 through milk consumption is a public concern. The purpose of this study was to determine the AFM1 exposure through liquid milk consumption for adult consumers in Tehran. Forty-five samples including raw, pasteurized and UHT milk samples were collected from markets in different cities of Tehran province in January and February 2017. The AFM1 was determined by HPLC method after immunoaffinity column clean up. Also, the milk intake was calculated using household budget survey. Finally, the daily intake of AFM1 through milk consumption was estimated using a deterministic approach. From total 45 samples, AFM1 was detected in 36 (80%) samples, although none of the analyzed samples were exceeded Iran legal limit of 0.1 µg/kg. On the basis of the average milk intake, the mean daily exposure to AFM1 was estimated between 0.03 ng/ Kg BW per day (lower bound estimate) and 0.06 ng/ Kg BW per day (upper bound estimate) and the 95th percentile daily exposure was calculated at 0.14 ng/ Kg BW per day. According to these values, it should be expected adults of Tehran population are not exposed to a significant risk of Hepatocarcinoma associated with AFM1 intake through milk consumption.
    Keywords: Aflatoxin M1, Milk, Exposure assessment, Tehran, Iran
  • Ehsan Motaghi, Valiollah Hajhashemia, Parvin Mahzounib, Mohsen Minaiyan* Pages 1341-1350
    Ulcerative colitis is chronic and recurrent disease of the gastrointestinal tract with uncertain etiology and incomplete treatment options. N-methyl-d-aspartate (NMDA) receptor suppression has shown anti-inflammatory effects in vitro and in vivo. The aim of present study was to evaluate the role of dizocilpine (MK-801), a noncompetitive NMDA receptor antagonist, on TNBS (trinitrobenzene sulfonic acid)-induced murine model of colitis. Dizocilpine (0.1, 1 and 5 mg/kg) was given to mice intraperitoneally from 24 hours before induction of colitis and daily thereafter for 4 days. Dexamethasone (1 mg/kg) was used as the reference drug. Colitis was induced by intracolonic administration of TNBSof TNBS/Ethanol (50/50 v/v, 40mg/kg). Animals were sacrificed 5 days after colitis induction and distal colons were examined macroscopically and microscopically. The colonic tissue level of pro-inflammatory cytokines including interleukin 1β (IL-1β), interleukin 6 (IL-6) and tumor necrosis factor-α (TNF-α) were assessed by ELISA. Myeloperoxidase (MPO) level was also measured in colon. Dizocilpine, particularly with intermediate dose of 1mg/kg significantly improved animal’s weight loss as well as macroscopic and microscopic signs of colitis, reduced colonic levels of IL-1β, IL-6, TNF-α and MPO activity. Hence, dizocilpine has significant protective effects in TNBS- induced colitis and NMDA suppression may be a new and effective therapeutic strategy in ulcerative colitis via decreasing in pro-inflammatory cytokine production.
    Keywords: Dizocilpine, MK-801, Ulcerative colitis, Mice, NMDA
  • Amirali Delnavaz Shahr, Fazel Nasuhi Pur* Pages 1351-1357
    Mexiletine as the first choice drug in myotonia treatment is a chiral sodium channel blocker clinically used in its racemic form. The phenolic structure of this drug, prompted us to design its novel calix[4]arene-based cluster in a chalice-shaped structure. Therefore, the present study reports the synthesis and in-vitro anti-myotonic activity of the chalice-shaped cluster of mexiletine (namely calixmexitil) in comparison to its simple drug unit (mexitil) as the reference medication. The synthetic route included chemical modification of the calix[4]arene structure by grafting four 2-aminopropoxy moieties at the lower rim of the scaffold. Electrophysiological tests were performed for the determination of test compounds abilities to act as sodium channel blockers in inhibiting sodium currents (in use-dependent manner) in single skeletal muscle fibers. The experimental results showed an amplified (10-fold) potency in producing phasic block as an indication of the anti-myotonic activity and improved (3-fold) potency in producing use-dependent block for the cluster (calixmexitil) in relation to its monomer (mexiletine). The potency in producing phasic block and use-dependent block are two main factors to describe dose range, drug affinity, and side effects of an anti-myotonic agent. Therefore, compared to mexiletine, calixmexitil with these improved factors can be considered as a “selective” anti-myotonic agent with low dose range. These improved pharmaceutical effects are maybe attributed to clustering effect and improved interaction of four impacted mexiletine units of the cluster with the sodium channels’ structure in skeletal muscle fibers.
    Keywords: Mexiletine, Cluster, Calixarene, Antimyotonic activity, Amplify, Sodium channel blocker
  • Saeideh Saadat, Mahsa Yasavoli, Zahra Gholamnezhad, Mohammad Reza Aslani, Mohammad Hossein Boskabady* Pages 1358-1370
    Crocin, a component of saffron, showed hypotensive which is perhaps due to vascular smooth muscle relaxant effect. The relaxant effects of saffron on tracheal smooth muscle also could be due to its constituent, crocin. In the present study, the relaxant effects of crocin and its possible mechanisms on rat tracheal smooth muscle were investigated. The relaxant effects of three cumulative concentrations of crocin (30, 60, and 120 μM) or theophylline (0.2, 0.4, 0.6 mM) as positive control was examined on pre-contracted tracheal smooth muscle by methacholine or KCl in non-incubated or incubated conditions with different agents including atropine, chlorpheniramine, indomethacin, diltiazem, glibenclamide and propranolol. In non-incubated tracheal smooth muscle, crocin showed significant relaxant effects on KCl induced muscle contraction (p
    Keywords: Crocin, Smooth muscle, Trachea, Receptors, Adrenergic, Muscarinic
  • Mehrnaz Rezaei, Hossein Ghafoori*, Mahmood Reza Aghamaali, Mostafa Shourian Pages 1371-1379
    The present study was designed to investigate the inhibitory effect of 2,4 bis-[(4-ethoxyphenyl)azo] 5-(3-hydroxybenzylidene) thiazolidine-2,4-dione (TZD-OCH2CH3) on the cyclo-oxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in RAW 264.7 cells. The effects of TZD-OCH2CH3 on COX-2 and iNOS mRNA expression in LPS-activated RAW 264.7 cells were detected by real time PCR. Also, to understand structure and substrate specificity, we have utilized molecular docking simulations (AutoDock Vina) and the active residues in the binding pocket were determined from COX-2 and iNOS. The treatment of RAW 264.7 cells with TZD-OCH2CH3 significantly inhibited LPS-induced COX-2 mRNA expression, corresponding to 46.1% and 61.06% at 30 and 60 μg/mL, respectively. The present study revealed that the TZD-OCH2CH3 had a little effect on iNOS mRNA expression. Meanwhile, the TZD-OCH2CH3 also could inhibit the production of NO compared to single LPS-stimulated cell. According to the results obtained, TZD-OCH2CH3 dramatically suppressed lipopolysaccharide (LPS) induced nitric oxide (NO) production after 24 h, in a concentration-dependent manner with an IC50 of 65 μg/mL. Our data suggest that TZD-OCH2CH3, as a functionally novel agent, inhibits the inflammatory pathway via suppression of COX-2 mRNA expression and also by the inhibition of the iNOS activity. Therefore, this compound could be suggested as a novel therapeutic strategy for inflammation-associated disorders.
    Keywords: Thiazolidinedione, Cyclo-oxygenase-2, Nitric Oxide Synthase, Nitric oxide, RAW 264.7 cells
  • Mohabbat Jamhiri, Farzan Safi Dahej, Akram Astani, Seyed Hasan Hejazian, Zeinab Hafizibarjin, Mojtaba Ghobadi, Ali Moradi, Arezu Khoradmehr, Fatemeh Safari* Pages 1380-1394
    Hypertension-induced left ventricular hypertrophy is the most important risk factor for heart failure. This study aimed at investigating the effects of monoterpenoid phenol, carvacrol, on myocardial hypertrophy using both in-vivo and in-vitro models. Male Wistar rats were divided into the control (Ctl), un-treated hypertrophy (H), and carvacrol-treated hypertrophy groups (25, 50 and 75 mg/kg/day, Car+H). In the hypertrophy groups animals underwent abdominal aorta banding. Blood pressure (BP) was recorded via carotid artery cannulation. TUNEL assay and Masson’s trichrome staining were used to assess apoptosis and fibrosis, respectively. The 2-2-diphenyl 1-picril-hydrasil)DPPH( radical scavenging activityand malondialdehyde (MDA) level were estimated by biochemical tests. In in-vitro study H9c2 cardiomyoblasts were treated with angiotensin II (Ang II) to promote hypertrophy. Cell size was measured using crystal violet staining. Gene expression was evaluated by real-timeRT-PCR technique. In the carvacrol-treated rats BP, heart rate, and heart weight to the body weight ratio were significantly decreased. In-vitro study showed that H9c2 cell size was significantly reduced compared to Ang II-treated cells. Both in-vivo and in-vitro studies demonstrated that carvacroldecreased atrial natriuretic peptide )ANP( mRNA level significantly (vs. H groups). The number of apoptotic cells increased in Hgroup, while it was decreased in the Car50+H and Car75+H. In Car+H groups, in comparison with H group, the serum concentration of MDA was decreased and DPPHwas increased significantly. Our findings demonstrated that carvacrol decreases hypertrophy markers in in-vivo and in-vitro models of hypertrophy.
    Keywords: cardiac hypertrophy, Carvacrol, Rat, H9c2 cells, Blood pressure, Antioxidant, Apoptosis
  • Leila Etemad, Mahdieh Zamani, Mehrdad Iranshahi, Ali Roohbakhsh* Pages 1395-1402
    It is believed that some pitfalls in the treatment of epilepsy including serious side effects of medications and drug resistance may be resolved by natural compounds. Auraptene is a natural coumarin that is found in many plants, particularly citrus peel. We hypothesized that auraptene might have anticonvulsant properties. Kindling was induced by repeated intraperitoneal (IP) injections of pentylenetetrazol (PTZ, 35 mg/kg) with two-day intervals for 24 days in male mice. Three groups received IP injections of auraptene (12.5, 25, and 50 mg/kg). Three control groups received vehicle, diazepam (3 mg/kg, IP), and vitamin E (150 mg/kg, IP). Seizure-related behaviors were recorded for 30 min after PTZ injection. Moreover, malondialdehyde and reduced glutathione (GSH) were measured in the brain. The results indicated that auraptene at the dose of 12.5 mg/kg and vitamin E prolonged the latency to stage 2 of seizures (P< 0.01). Auraptene at the doses of 25 mg/kg and 50 mg/kg, prolonged the latency to stage 4 (P< 0.01) and reduced stage 5 duration of seizures (P< 0.01). All doses of auraptene reduced median of seizure scores (P< 0.01). The kindled control group had MDA levels similar to intact animals but had a lower concentration of GSH (P< 0.001). None of the tested compounds changed the malondialdehyde concentration significantly. However, auraptene at the dose of 50 mg/kg and vitamin E increased GSH levels (P< 0.05). The results suggested that auraptene had anticonvulsant effects in PTZ-induced chemical kindling that was mediated by mechanisms other than the antioxidant effect of auraptene.
    Keywords: Pentylenetetrazol, Auraptene, Chemical kindling, Oxidative stress, Seizure
  • Farideh Bahrami, Alireza Asgari, Narges Hosseinmardi, Mahyar Janahmadi* Pages 1403-1418
    Alzheimer’s disease (AD) is undoubtedly one of the serious and growing public health challenges in the world today. There is an unmet need for new and effective preventative and therapeutic treatment approaches for AD, particularly at early stages of the disease. However, the underlying mechanism against Aβ-induced electrophysiological alteration in cultured hippocampal pyramidal neurons  is still not fully understood. This study investigated  the impacts of activation and inhibition of PPAR-γ/δ on the Aβ-induced functional toxicity, which occured before cell death, using patch clamp technique. Findings demonstrated that Aβ treatment alone altered the normal electrophysiological properties and reduced the Ca2+ channel currents in primary cultured hippocampal pyramidal neurons without any major changes either in cell structure, as evidenced by electron microscope examination, or cell viability. Rosiglitazone (30 μM), a potent PPAR-γ activator, when co-treated with Aβ (100 nM) prevented almost completely the induction of function toxicity of Aβ, as evidentiated by restored normal appearing electrophysiological properties. Inhibition of PPAR- γ/δ by FH535 (15 μM), an inhibitor of both Wnt/beta-catenin signaling and PPAR- γ and δ activity, when applied in combination of Aβ not only worsen the toxic electrophysiological effects of Aβ on firing frequency, membrane resistance and cell viability, but also even preserved the suppressive effect of Aβ on Ca2+ channel current when compared to control condition. Overall, these findings suggest that PPAR-γ activation could be a potential candidate to prevent the functional changes induced by low concentration of Aβ which may possibly occur in neurons during early stages of AD.
    Keywords: Alzheimer’s disease, Neurodegenerative diseases, Ca2+ channel current, PPAR-γ, Amyloid Beta (Aβ), Hippocampal pyramidal neurons
  • Shahram Darabi*, Ali Noori, Zadeh, Hojjat, Allah Abbaszadeh*, Farzad Rajaei, Salar Bakhtiyari Pages 1419-1428
    Trehalose, as a natural disaccharide, is known as an autophagy inducer. The neuroprotectiveeffects of trehalose in the rat model of Parkinson′s disease were the aim of the present study.Parkinson′s disease model was induced by injecting 6-hydroxydopamine (6-OHDA) in thestriatum of male Wistar rats. Apomorphine-induced behavior and substantia nigra neuronalcounts were applied to evaluate the neuroprotective effects of trehalose. The autophagy wasstudied using the expression of p62 and LC3II/LC3I ratio. In addition, the antioxidant effectsof trehalose were assessed by analyzing the levels of nuclear factor (erythroid-derived 2)-like2 (Nrf2) and also glutathione reductase (GR), glutathione peroxidase (GPx) and Catalase(CAT) enzymes. Moreover, the levels of 3, 4-dihydroxyphenylacetic acid (DOPAC) anddopamine (DA) were assessed.The behavioral test showed that trehalose in the treatment groupreduced the damage to the substantial nigra dopaminergic neurons, which was characterizedby improved motor and reduced rotations in the treatment group as compared with the lesiongroup. In the histological examinations of the treatment group, trehalose prevented thedestruction of dopaminergic neurons. Trehalose treatments increased autophagy (high LC3II/LC3I ratio) and the expression of the p62 protein as well. Through p62-dependent manner,it led to increased nuclear translocation of Nrf2 transcription factor and elevated expressionof downstream antioxidant enzymes, such as GR, GPx, and CAT, restoring DA and DOPACcontents of the cells. In the current study, trehalose simultaneously protects substantia nigradopaminergic cells by activating both non-canonical p62/SQSTM1-Keap1-Nrf2 and autophagypathways.
    Keywords: Autophagy, Non-canonical Nrf2 pathway, p62, Parkinson′s disease, Trehalose
  • Azadeh Gholaminejad, Hamid Gholamipour, Badie, Mohammad Nasehi, Nasser Naghdi* Pages 1429-1444
    Prefrontal cortex (PFC) is involved in multiple functions including attentional , spatial orientation, short and long-term memory. Our previous study indicated that microinjection of testosterone in CA1 impaired spatial learning and memory. Some evidence suggests that impairment effect of testosterone is mediated by GABAergic system. In the present study, we investigated the interaction of testosterone (androgenic receptor agonist) and bicuculline (GABAA receptor antagonist) on spatial learning and memory in the prelimbic (PL) of rats. Cannulae were bilaterally implanted into the PL region of PFC and drugs were daily microinjected for two minutes in each side. There are 4 experiments. In the first experiment, three groups sham operated (solvent of testosterone, bicuculline, testosterone plus bicuculline). In the second experiment, different doses of testosterone (40, 80 μg /0.5 μl DMSO/each side) were injected into the PL before each session. In the third experiment, intra PL injections of bicuculline (2, 4 μg/0.5 μl DMSO/each side) were given before every session. In the last experiment, testosterone (80μg/0.5 μl DMSO/each side) along with bicuculline (2 μg/0.5 μl DMSO/each side) was injected into the PL. The results showed there is no differences between control and sham operated group. Testosterone 80 μg and bicuculline 2 μg, each given separately, and also in combination increased escape latency to find the platform compared to the sham operated and cause to impaired spatial learning and memory. It is shown that intra PL microinjection of bicuculline after testosterone treatment could not rescue the spatial learning and memory impaired induced by testosterone.
    Keywords: Prelimbic, Spatial memory, Testosterone, Bicuculline, GABAA receptor, Morris Water Maze
  • Nasim Manouchehri, Fariba Khodagholi, Leila Dargahi, Abolhassan Ahmadiani* Pages 1445-1455
    Preconditioning (PC) as a protective strategy against noxious insults can decline cell death and apoptosis. It has been approved that mitochondria play a key role in PC mechanism. The critical role of complex I (CI) in oxidative phosphorylation machinery and intracellular ROS production, particularly in the brain, accentuates its possible role in PC-induced neuroprotection. Here, differentiated PC12 cells were preconditioned with ultra-low dose LPS (ULD, 3 μg/mL) prior to exposure to high concentration of LPS (HD, 750 μg/mL). Our results showed that HD LPS treatment reduces cell viability and CI activity, and intensifies expression of cleaved caspase 3 compared to the control group. Intriguingly, PC induction resulted in enhancement of cell2viability and CI activity and reduction of caspase3 cleavage compared to HD LPS group. In order to explore the role of CI in PC, we combined the ULD LPS with rotenone, a CI inhibitor. Following rotenone administration, cell viability significantly reduced while caspase3 cleavage increased compared to PC induction group. Taken together, cell survival and reduction of apoptosis followed by PC can be at least partially attributed to the preservation of mitochondrial CI function.
    Keywords: Mitochondrial complex I, Preconditioning, Neuroinflammation, Neuroprotection, Mitochondria
  • Reza Jahani, Faraz Mojab, Arash Mahboubi, Azadeh Nasiri, Armin Tahamtani, Mehrdad Faizi* Pages 1456-1465
    Antidepressant-like activity of T. kotschyanus has been recently reported by scientists but insufficient attention has ‎so far been devoted to T. kotschyanus, and there is a lack of information on the other neurobehavioral effects and ‎side effects of this species. In the current study, the anticonvulsant, anxiolytic and sedative-hypnotic, effects of ‎Thymus kotschyanus extract on male NMRI mice were evaluated using pentylenetetrazole, maximal electroshock, ‎elevated plus maze, and pentobarbital-induced sleeping tests. Since phenolic compounds and flavonoids have main ‎roles in pharmacological effects of most plant extracts, the phenolic and flavonoid contents of the extract were ‎measured with Folin-Ciocalteu and AlCl3 reagents. Acute toxicity, passive avoidance, and open field tests were ‎carried out to assess the toxicity of the extract. To find out the possible mechanism of action, flumazenil as the ‎specific GABAA receptor antagonist was used. Anticonvulsant and hypnotic effects of the extract were observed at ‎‎400 and 600 mg/kg. The extract at the dose of 200 mg/kg revealed significant anxiolytic effects, but it did not show ‎any adverse effects on learning and memory at all the tested doses. Results of this study indicate that Thymus ‎kotschyanus extract has anticonvulsant‎, anxiolytic and hypnotic effects, which are likely related to the ability of ‎some phenolic compounds to activate α1-containing GABAA receptors but more experiments still need to be carried ‎out in order to find the exact mechanism, active component, and the toxicity of the Thymus kotschyanus extract. ‎
    Keywords: Thymus kotschyanuse, GABA-A receptors, Phenolic content, Epilepsy, Insomnia, memory, Mice
  • Azam Ghanbari, Maman, Forouzan Ghasemian, Roudsari, Shayan Aliakbari, Hamid Gholamipour, Badie*, Fariba Khodagholi, Fatemeh Shaerzadeh, Mahtab Daftari Pages 1466-1476
    Entorhinal cortex (EC) is one of the first Entorhinal cortex (EC) is one of the first cerebral regions affected in Alzheimer’sdisease (AD). The pathology propagates to neighboring cerebral regions through a prion-likemechanism. In AD, intracellular calcium dyshomeostasis is associated with endoplasmicreticulum (ER) stress. This study was designed to examine hippocampal ER stress followingEC amyloidopathy. Aβ1-42 was bilaterally microinjected into the EC under stereotaxic surgery.Rats were daily treated with 30 μg of isradipine, nimodipine, or placebo over one week.Passive avoidance and novel object recognition (NOR) tasks were performed using shuttle boxand NOR test, respectively. GRP78/BiP and CHOP levels were measured in the hippocampaldentate gyrus (DG) by western blot technique. The glutathione (GSH) level and PDI activitywere also assessed in the hippocampus by colorimetric spectrophotometer. Aβ treated groupdeveloped passive avoidance and novel recognition memory deficit compared to the controlgroup. However, treatment with calcium channel blockers reversed the impairment. BiP andCHOP level increased in the hippocampus following amyloidopathy in the EC. PDI activityand GSH level in the hippocampus decreased in the Aβ treated group, but calcium channelblockers restored them toward the control level. In conclusion, memory impairment due to ECamyloidopathy is associated with ER stress related bio-molecular changes in the hippocampus,and treatment with L-type calcium channel blockers may prevent the changes and ultimatelyimprove cognitive performance.cerebral regions affected in AD. Intracellular calcium buffering capacity is disrupted in the dentate gyrus (DG) following EC amyloidopathy. This study was designed to examine hippocampal endoplasmic reticulum (ER) stress following EC amyloidopathy. Aβ1-42 was bilaterally microinjected into the EC under stereotaxic surgery. Rats were daily treated with 30 μg of isradipine, nimodipine or placebo over one week. Passive avoidance and novel object recognition (NOR) tests were performed. GRP78/BiP and CHOP levels were measured in the hippocampal DG. The glutathione (GSH) level and PDI activity were also assessed in the hippocampus. Aβ treated group developed passive avoidance and novel recognition memory deficit compared to the control group. However, treatment with calcium channel blockers reversed the impairment. BiP and CHOP level increased in the hippocampus following amyloidopathy in the EC. PDI activity and GDH level in the hippocampus were decreased in the Aβ treated group, but calcium channel blockers restored them toward the control level. In conclusion, memory impairment due to EC amyloidopathy is associated with ER stress related bio-molecular changes in the hippocampus, and treatment with L-type calcium channel blockers may prevent the changes and ultimately improve cognitive performance.
    Keywords: Alzheimer disease, entorhinal cortex, endoplasmic reticulum stress, calcium channel blockers, protein disulfide-isomerases
  • Didi Nurhadi Illian, Poppy Anjelisa Zaitun Hasibuan, Sumardi Sumardi, Arif Nuryawan, Ridha Wati, Mohammad Basyuni* Pages 1477-1487
    Colorectal cancer is the third most common cancer world wide and has been occurred more in developing regions. The use of conventional chemotherapy agents may lead to various adverse effects. Therefore, it is required to find the potential drug for anticancer from alternative source of natural product including mangrove plants. The present study was conducted to determine the anticancer activity of polyisoprenoids from Avicennia alba Blume. leaves (PAL) in WiDr cells. Cell cycle inhibition, apoptosis activity, and suppression of cyclooxygenase-2 (COX-2) were also evaluated. The anticancer activity of PAL was determined by observing the activity of these compounds against WiDr cells using the [3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazolium bromida] MTT assay. Inhibition of the cell cycle and increased apoptosis were analysed by flowcytometry. Suppression of COX-2 was analysed using immunocytochemistry. PAL exhibited anticancer activity against WiDr cells with an IC50 of 173.775 μg/mL. Cell cycle analysis revealed that the inhibition occurred in the G0-G1 phase, and apoptosis occurred in the early apoptosis phase. Furthermore, the result of an analysis of COX-2 expression showed that PAL enabled the suppression of COX-2 expression. PAL can be used as anticancer agents against WiDr colon cancer cells. However, in-vivo studies is required to confirm the in-vitro finding of the anticancer activity of polyisoprenoid extract.
    Keywords: Polyisoprenoids, Avicennia alba, Apoptosis, Cell Cycle, COX-2
  • Mohammad Amin Rajizadeh, Hamid Najafpour*, Mitra Samareh Fekri, Farzaneh Rostamzadeh, Elham Jafari, Mohammad Abbas Bejeshk, Yaser Masoumi, Ardakani Pages 1488-1498
    Aims
    To investigate the effect of Myrtenol, the active ingredient of Myrtle, on the oxidant and anti-oxidant indices and cytokines in the allergic asthma.
    Methods
    Allergic asthma was induced by ovalbumin (OVA) sensitization and inhalation in four groups of rats; Control, Asthma, Asthma+Dexamethasone and Asthma+Myrtenol. Myrtenol (50mg/kg) or Dexamethasone (2.5mg/kg) was administered intraperitoneally for 7 consecutive days after OVA inhalation. At the end, histopathological parameters and interleukins (Interleukin-10 (IL10), Interferon gamma (IFN-γ) , interleukin-1β (IL-1β), Tumor Necrosis Factor α (TNF-α)) and oxidative stress biomarkers, Malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GPX) in the lung and serum were measured by hematoxylin & eosin staining and ELISA method respectively.
    Results
    Myrtenol reduced the pathological changes in the lungs and airway endothelium (P<0.01), (P<0.5). The level of IL-1β (P < 0.05) and MDA in the serum and lung tissue (P < 0.01), (P < 0.05), and also the level of TNF-α (P < 0.05) in the lung tissue decreased in the Myrtenol group compared to the asthma group. Myrtenol increased the level of IL-10 ( P < 0.05) and the activity of GPX in the lung tissue and serum (P < 0.001).
    Conclusion
    Myrtenol may improve asthma by increasing the ratio of antioxidants to oxidants and reducing the ratio of pro-inflammatory to anti-inflammatory interleukins in the lung. Myrtenol is presented as a potent herbal medicine ingredient for the treatment of asthma.
    Keywords: Allergic asthma, Myrtenol, malondialdehyde, Superoxide dismutase, glutathione peroxidase, Cytokines
  • Kun Cao, Wei Qian, Yi Xu*, Zhen Zhou Pages 1499-1507
    Sesquiterpenes lactones including costunolide and dehydrocostus lactone, were isolated from Saussurea lappa roots, which had exhibited a wide range of biological activities such as anti-cancer, anti-inflammatory, antiulcer, and immunomodulatory activities. High-speed counter-current chromatography (HSCCC) was applied for the rapid preparative isolation of sesquiterpenes lactones from Saussurea lappa roots. A solvent optimization method for HSCCC was presented, i.e. the separation factors of compounds after the partition coefficient (K) of solvent system should be investigated. Using this method, 150 mg of costunolide, 140 mg of dehydrocostus and 15 mg of 10-methoxy-artemisinic acid with purities of 95%, 98%, and 98% were obtained within 150 min. The structures of three compounds were identified by mass spectrum (MS) and nuclear magnetic resonance (NMR) spectroscopy. These results offered an efficient strategy for separation of potentially health-relevant phytochemicals from Saussurea lappa roots.
    Keywords: Saussurea lappa Roots, Sesquiterpenes lactones, Select solvent system, HSCCC, Orthogonal test
  • Shiren Deng, Linbo Xia*, Xiamin Zhu, Jiang Zhu, Mingchen Ca, Xin Wang Pages 1508-1515
    An efficient and rapid affinity-based screening method for directly fishing out natural alpha-glucosidase inhibitors from Cyperus. rotundus extract by using immobilized enzyme technology combined with UHPLC-QTOF MS analysis was established. As a results, without time-consuming and laborious isolation workload and false positive interference, five natural alpha-glucosidase inhibitors were successfully recognized and identified from only 400 uL of C. rotundus extracts within only a couple of hours, which suggested that the screening method was rapid, economical, sensitive and feasible. In addition, the captured compounds were isolated and characterized as stilbenoids oligomers, and were proved to be strong alpha-glucosidase inhibitors by inhibitory assay in vitro. Among them, 3 stilbenoids trimers were reported to be potent α-glucosidase inhibitors for the first time. This method could be modified and have the potential for rapidly screening of active compounds extracts against some new targets by immobilizing some other biomacromolecules.
    Keywords: Affinity fishing, alpha-glucosidase inhibitors, immobilized enzyme, Cyperus rotundus, Stilbenoid timers, UHPLC-QTOF MS
  • Songül Karakaya*, Duygu Şimşek, Hilal Özbek, Zühal Güvenalp, Nurten Altanlar, Cavit Kazaz, Ceyda Kılıc Pages 1516-1529
    Ferulago species have been utilized since ancient times as digestive, sedative, aphrodisiac, along with in salads or as a spice due to their special odors. The study reports isolation and characterization of bioactive compounds of Ferulago pachyloba (F. pachyloba), Ferulago trachycarpa (F. trachycarpa), Ferulago bracteata (F. bracteata), and Ferulago blancheana (F. blancheana) via bioassay guided fractionation and isolation process. The structures of compounds were elucidated by detailed spectroscopic analyses. They were also assessed for their activities at 1000-31.25 µg/mL concentrations by microbroth-dilution methods. Antimicrobial activity of aqueous, methanol extracts and dichloromethane, ethyl acetate, n-butanol and aqueous residue fractions of methanol extracts from aerial parts and roots of species along with isolated compounds [osthole, imperatorin, bergapten, prantschimgin, a new coumarin peucedanol-2′-benzoate, grandivitinol, suberosin, xanthotoxin, felamidin, marmesin, umbelliferone, ulopterol and a sterol mixture consisted of stigmasterol, β-sitosterol] were evaluated. Antimicrobial effect has been seen against Gram-negative, Gram-positive bacteria, and a yeast C. albicans at a concentration between 31.25 and 62.5 μg/mL. Especially, C. albicans (MIC = 31.25 μg/mL) was the most inhibited microorganism. Moreover, growth of P. aeruginosa, B. subtilis, E. coli,and S. aureus were inhibited at 62.5 μg/mL MIC values. Among tested samples prantschimgin  and dichloromethane fraction of aerial parts from F. pachyloba showed the best activity against C. albicans (MIC = 31.25 μg/mL). However, among aqueous extracts and residue fractions, only F. blancheana aerial parts, F. trachycarpa aerial parts and roots and F. bracteata roots showed activity against C. albicans. Among microorganisms E. coli was found to be the least affected.
    Keywords: Antimicrobial, Coumarin, Ferulago, Peucedanol-2′-benzoate, Prantschimgin
  • Mehrnoosh Sedighi, Mahdieh Faghihi, Mahmood Rafieian, Kopaei, Bahram Rasoulian, Afshin Nazari* Pages 1530-1542
    Abstract Melissa officinalis has antioxidant and anti-inflammatory activities and is used in various diseases. Aim of the study: We investigated the role of M. officinalis extract (MOE) against ischemia-induced arrhythmias and heart injury after five days of reperfusion in an in-vivo rat model of regional heart ischemia. The leaf extract of M. officinalis was standardized through HPLC analysis. Adult male rats were subjected to 30 min of ischemia by occlusion of the left anterior descending coronary artery followed by 5 days of reperfusion. The rats were randomized to receive vehicle or M. officinalis as follows: group I served as saline control with ischemia, groups II, III and IV received different doses of MOE- (intragastric, 25, 50 or 100 mg/kg respectively), by oral gavage daily for14 days prior to ischemia Administration of M. officinalis significantly improved ischemia/reperfusion (I/R)-induced myocardial dysfunction by reduction of infarct size, episodes of ventricular tachycardia(VT) and ventricular ectopic beat(VEB and duration of VT as compared to control group, stabilized ST segment changes, QTc shortening, increased the R and T wave amplitudes and increased the heart rate during ischemia, increased the serum superoxide dismutase (SOD) activity and decreased serum lactate dehydrogenase (LDH), Troponin I (CTnI) and malondialdehyde (MDA) levels, 5 days after reperfusion. MOE-100mg/kg was the effective dose. Cinamic acid (21.81±1.26 mg/gr) was the main phenolic compound of plant sample. The ethanol extract of M. officinalis was observed to exhibit cardioprotective effects against I/R injury, probably due to antioxidant properties.
    Keywords: Melissa officinalis, Ischemia, Reperfusion injury, Heart, Rat
  • Solmaz Asnaashari, Abbas Delazar, Elham Safarzadeh, Hamed Tabibi, Saeed Mollaei, Ali Rajabi, Parina Asgharian* Pages 1543-1555
    Scrophularia atropatana (S. atropatana), an Iranian plant belonging to the family of Scrophulariaceae, was assigned for its chemical compositions and biological activities of essential oil (EO) and extracts of the aerial parts of the plant during the flowering stage. Combination of GC-MS and GC-FID was assessed for analyzing the chemical compositions of the EO from the aerial parts of S. atropatana. Furthermore, Brine shrimp lethality test and DPPH assay were performed to evaluate general toxicity and free-radical-scavenging properties, respectively. Furthermore, anti-proliferative and antimicrobial activities were assessed by MTT assay and disc diffusion methods correspondingly. Additionally, all the potent samples (extracts) and its fractions in the MTT assay were further studied for the presence of various compounds by GC-MS apparatus. MeOH extract and 40% sep-pak fraction indicated high amounts of total phenolic (TPC), total flavonoid content (TFC), and antioxidant properties. In the case of general toxicity, among the extracts, dichloromethane (DCM) extract showed noticeable effect. Furthermore, DCM extract was indicated potent ability to eliminate breast tumor cells and minimum efficacy on normal cells. Anti-microbial activity of all samples was ignorable. The potent extracts and fractions which had more anti-proliferative activity were further elucidated by GC-MS and showed high amounts of Alkanes and fatty acids. In the case of EO constituents, non-terpenoids were the major compounds.To sum up, it seems BSLT could be a good preliminary approach for evaluating the cytotoxicity in MCF-7 cell line. Additionally, antioxidant activity, TPC, and TFC contents of all samples were in consistent with each other.
    Keywords: Scrophularia atropatana, phytochemical analysis, Biological activities, GC-MS analysis, MTT assay
  • Rui Zhao*, Bing Qiu Maste, Baoling Ma Master, Yaping Cai Pages 1556-1565
    Depressive disorder will be the second highest disease burden worldwide, which will impair life quality, reduce productivity, and increase disability and mortality. LBP is the main active fraction purified from Lycium barbarum. The aim of this study was to evaluate the potential therapeutic effects of LBP on depressive mice induced by reserpine, as well as the relevant mechanisms. The antidepressant effect of LBP was investigated by open field test (OFT), forced swimming test (FST), tail suspension test (TST) and antagonism of reserpine hypothermia and ptosis in mice. In addition, we examined the oxidative status and antioxidation power of striatum in both control and depressive mice with or without LBP treatment. To explore the mechanism of LBP on regulating antioxidants in the depressive mice, we detected the expression level of Bcl-2 and PARP in striatum of mice by western blotting. The results showed that administration with LBP for 4 consecutive weeks significantly increased locomotor activity, reduced the duration of immobility, and antagonized hypothermia and ptosis in mice induced by reserpine. Also, LBP treatment was able to reduce the lipid peroxidation (LPO) production, and enhance the antioxidation effect of the striatum in depressive mice. Furthermore, LBP inhibited the decreased extent of the apoptotic suppressors, Bcl-2 and PARP, which were markedly decreased after treatment with reserpine. The above results indicated that LBP possess antidepressant activities, probably via its powerful antioxidative properties and then decreased the apoptosis of striatum neuron.
    Keywords: Lycium barbarum, Polysaccharide, depressive disorder, Antioxidation, Apoptosis
  • Leila Zarandi, Miandoab, Mohammad, Amin Hejazi*, Mohammad, Bager Bagherieh, Najjar, Nader Chaparzadeh Pages 1566-1579
    During recent years, there was growing demand in using microalga valuable products such as β-carotene in health care. β-Carotene has anti-cancer and anti-aging properties for human. In Dunaliella salina cells, β-carotene has a major protecting role for biomolecules, when the production of reactive oxygen species elevated. In the present study, we investigated the influence of the four most effective factors (light intensity, temperature, nitrate and salinity concentration) and their interactions on the β-carotene production and the total chlorophyll/β-carotene ratio in low light adapted D. salina cells. Box-Benken design and response surface methodology (RSM) was used for this purpose and optimization of the factor levels. Two models were developed to explain how β-carotene productivity and the total chlorophyll/β-carotene ratio depend on the stress factors. Among the four stress variables for β-carotene production, light intensity was stronger than the others. Meanwhile, interaction between light intensity and salt concentration exhibited the most important effect on the total chlorophyll/ β-carotene ratio. The predicted optimal conditions for maximum β-carotene productivity and minimum total chlorophyll/β-carotene ratio were derived from the fitted model in 200 µmol photons m-2s-1 light intensity, 25ºC, 0.9 mM nitrate and 3.8 M NaCl. When the predicted condition was tested experimentally, the close results were captured. This suggests that overproduction of β-carotene in D. salina under certain conditions depends on used light intensity for preadaptation. The step-wise manner applying of stresses may act as a benefit strategy to β-carotene overproduction.
    Keywords: Dunaliella salina, β-Carotene production, optimization, RSM, Pre adaptation
  • Alireza Mokhtari, Kumarss Amini* Pages 1580-1594
    Pseudomonas aeruginosa is an important multi-drug resistant (MDR) opportunistic bacterium. 102 strains of Pseudomonas aeruginosa equally isolated from human and cow milk were subjected to Multiplex-PCR for detection of ESBLs and exoenzymes of U, T, S, OprI, and OprL, Integrons class A encoding genes and genotyping by the ERIC-PCR and PFGE methods. The disc diffusion and E-test based on CLSI (Clinical and Laboratory Standards Institute) were performed to identify the antibiotics’ resistant strains. Exotoxin A encoding gene was detected in more than 90% of the studied strains, exoenzyme S prevalence in isolated samples from animal (cow milk) was negative and the frequency of Exo Y, Exo T, and Exo U were 25%, 68.6%, and 68.6%, respectively. The frequency of VEB and GES encoding genes in human strains were detected as 3.9% and 0 by Multiplex-PCR, respectively. The highest resistance was seen to Ampicillin and Cefepime (100%) while the lowest was observed to Amikacin (80.3%). E-Test results on human and animal strains showed complete resistance to Meropenem and Ampicillin, respectively. Dendrogram of ERIC-PCR method on human isolated samples revealed 22 different groups. Frequency of Integron I encoding gene was detected as 21.5% and 1.96% in human and animal strains, respectively. In general, the present study showed the high value of genetic diversity among isolates from animal and human samples with different progenitors, but the clones classified in one cluster revealed the same source of infection.
    Keywords: Pseudomonas aeruginosa, Multiplex-PCR, ESBLs, PFGE, ERIC-PCR
  • Nader Dilmaghani*, Nadereh Alani, Sepideh Fazeli Dehkordi Pages 1595-1601
    Recent studies suggest a relationship between zinc deficiency and inflammation. In the present study, we studied the effect of oral zinc supplementation on clinical improvement of chronic rhinosinusitis with nasal polyposis. In this single-blind randomized controlled trial, 44 patients with chronic rhinosinusitis with polyposis referring to ENT clinic of the Loghman Hakim hospital during 2013-2014 were randomly allocated in two groups. The treatment group (n = 28) was treated with a four-drug fixed-dose regimen (FD_FDR) consisting of oral dexamethasone (0.02 mg/kg), fluticasone nasal spray, fexophenadine 60 mg daily, montelukast 10 mg daily plus 220mg zinc sulfate capsules containing 55 mg elemental zinc, b.d., and the control group (n = 16) received the FD_FDR without supplemental zinc, for six weeks. After sixth week, two groups were compared regarding clinical outcomes based on theSNOT20 (Sinonasal outcome test) questionnaire, the general health questionnaire (SF12), the Lund-Mackay, and the Lund-Kennedy scoring systems. In the treatment group, serum zinc levels were significantly increased compared to those at the baseline (1.33 fold-increase; p = 0.0002). Within groups analysis revealed a significant reduction (p < 0.01) in LM and LK in both treatment (55% LM; 50% LK) and control groups (45% LM; 53% LK). Incontrast, between groups analysis revealed no significant differences in the LM and LK. The treatment group showed a mild superiority in general health improvement compared to that of the control group. Add-on therapy with supplemental zinc sulfate was not associated with significant improvement in patients with chronic rhinosinusitis with nasal polyposis (CRSwNP). The advantage of zinc supplementation on the general health improvement of the patients with CRSwNP requires further assessments.
    Keywords: Chronic rhinosinusitis, Nasal polyposis, zinc, Lund-Kennedy score, Lund-Mackay score
  • Neda Haji, Ali, Nili, Fariba Khoshzaban*, Mehrdad Karimi, Roja Rahimi, Elham Ashrafi, Ghaffari Reza, Ali Ghobadi, Mahmoud Jabarvand Behrouz Pages 1602-1611
    This study was designed to evaluate efficacy of a natural eye drop made of Plantago ovata mucilage versus placebo to improve dry eye symptoms. In a Randomized, double-masked, placebo-controlled clinical trial, sixty dry eye patients with ocular symptom and total OSDI score≥12 were randomly assigned to receive either a natural ophthalmic drop, made of Plantago ovata mucilage or placebo 4 times a day for 6 weeks. Patients were evaluated at first, weeks 4 and 6 of treatment. Evaluations of efficacy and safety were conducted based on Ocular Surface Disease Index questionnaire, Noninvasive tear film break-up time (NI-BUT) with keratograph, Schirmer test without anesthesia, osmolarity test and by monitoring adverse events. After 6 weeks, the result showed significant improvement in total OSDI score within (p
    Keywords: Dry eye disease, Persian medicine, Plantago ovata, Mucilage, Randomized controlled trial
  • Marya Kabiri, Mohammad Kamalinejad, Soodabeh Bioos, Mamak Shariat, Farnaz Sohrabvand* Pages 1612-1621
    Chamomile is a fascinating plant quoted in several traditional medicine texts, which hasbroad-spectrum pharmacological activity and medicinal uses. The aim of this study was toassess the efficacy of chamomile syrup in reducing serum prolactin in women with idiopathichyperprolactinemia. The study was a randomized, controlled clinical trial that was conductedon 56 women with idiopathic hyperprolactinemia for a study period of four weeks. Patientswere randomly enrolled in two parallel arms and were treated by chamomile syrup at a doseof 5 mL twice daily or cabergoline tablet orally at a dose of 0.25 mg twice weekly. Serumprolactin levels were measured at baseline and the end of the 4-week study period. Any reportof adverse events was also recorded. Results revealed that within the cabergoline group thereduction in the mean prolactin level was significantly greater than that of the chamomilegroup (p
    Keywords: Hyperprolactinemia, Matricaria chamomilla, Cabergoline, Prolactin, Complementary Medicine, Chamomile
  • Majid Zia, Behbahani, Hossein Niknahad, Javad Kojuri, Mahmood Salesi, Mojtaba Jafari, Khosro Keshavarz* Pages 1622-1631
    Background
    Acute myocardial infarction (AMI) is the leading cause of death throughout the world. One of the standard approaches to treatment of AMI is fibrinolysis. The study was conducted to evaluate the clinical efficacy of tenecteplase versus reteplase through network meta-analysis for AMI.
    Methods
    Randomized trials were comprehensively searched in PubMed, Scopus, Cochrane library, and Web of Science using appropriate strategies. Quality assessment was done for the papers. The primary and secondary end-points were mortality, TIMI grade 3 flow at 90 minutes, death or non-fatal stroke, infarction, total stroke and major bleeding. Odds ratios (OR) were computed (95% confidence intervals).
    Results
    After screening 27325 records, eight articles were included with total patients of 49875 to the meta-analysis. Indirect comparison of tenecteplase vs. reteplase showed no significant differences in the risk of mortality (OR= 0.98, p>0.05), TIMI grade 3 flow at 90 minutes (OR= 0.77, p>0.05), death or non-fatal stroke (OR= 1.04, p>0.05), infarction (OR= 1.11, p>0.05), total stroke (OR=2.71, p>0.05), and major bleeding (OR= 0.81, p>0.05) (all p>0.05).
    Conclusions
    Indirect comparison suggests similar efficacy and safety of tenecteplase and reteplase. Hence, the use of each one of the two medicines depends on price, facility, and accessibility of the medicine.
    Keywords: tenecteplase, Reteplase, alteplase, systematic review, Meta-Analysis, Efficacy, Safety
  • Seyed Massood Nabavi, Roya Abolfazli, Ali Etemadrezaei, Hamed Hosseini, Nahid Moradi, Sanaz Shahriari, Baharak Mehdipour, Babak Shekarchi, Akbar Soltanzadeh* Pages 1632-1638
    We compared the efficacy and safety of a biosimilar form of beta-interferon-1a (Actovex) versus the reference product in the treatment of relapsing remitting multiple sclerosis (RRMS). In a double blind, randomized phase 3 clinical trial, we evaluated 138 patients with RRMS that were allocated to receive the biosimilar medication and the reference treatment (30 μg intramuscular, weekly for one year). We investigated changes in EDSS, relapse rate and MRI changes within one year. In sixty-nine patients who were allocated to each arm and analyzed mean age and its standard deviation was 32.4 ± 8.8 and 31.5 ± 8 for the biosimilar medication and the reference arm respectively. One-year follow-up revealed a mean difference of 0.084 in EDSS (95% CI: 0.069-0.237) between the two groups in favor of the biosimilar medication. This value did not exceed the predefined non-inferiority margin of 0.1. There were no statistically significant differences in relapse rate and systemic and local adverse events of the two groups. The results show that the biosimilar interferon 1-a is non-inferior to the reference product in terms of efficacy while it demonstrates comparable safety. In conclusion the biosimilar interferon 1-a can be considered as an effective and safe alternative to the reference product due to lower cost and more availability.
    Keywords: Relapsing remitting multiple sclerosis, Beta interferon -1a, Biosimilar, Multiple sclerosis, Pharmacologic therapy
  • Hakimeh Zali, Ali Golchin, Masoumeh Farahani, Mohsen Yazdani, Mohammad Mehdi Ranjbar, Ali Dabbagh* Pages 1639-1647
    Accumulating evidence indicates that toll-like receptor 4 (TLR4) plays a critical role in promoting adaptive immune responses and are definitively involved in the expansion and maintenance of the neuropathic pain. However, the application of docking in virtual-screening in silico methods to drug discovery has some challenge but it allows us to make the directed and meaningful design of drugs for a target protein, which can be shortening and low costing the evolution and discovery of very promising lead new drugs. Nevertheless, in parallel with virtual screening methods, attendant developments in cell culture and in vivo studies must be achieved. In the present paper, we aimed to discover new drugs that have the ability to bind and inhibit TLR4 functions. So, after using the Pathway studio to investigate the biological pathways and protein interaction maps between TLR4 and neuropathy, we reported the application of the affinity-based approach of different pharmaceuticals; these agents contained all of the approved drugs; which could bind to Toll-like receptor 4 in blind high-throughput in silico screening. Our results demonstrated that among the primary list of 1945 retrieved compounds, 39 approved compounds could be the right candidate to perform a biological test in different in vivo and in vitro conditions and as a lead for further neurophysiological and neuropathological studies and treatment of neuropathic pain.
    Keywords: Approved drugs, docking, pathway studio, TLR4, neuropathic pain, virtual-screening