فهرست مطالب

Advanced Pharmaceutical Bulletin - Volume:9 Issue:3, 2019
  • Volume:9 Issue:3, 2019
  • تاریخ انتشار: 1398/05/26
  • تعداد عناوین: 20
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  • Fataneh Hashempour, Hedayat Hosseini*, Saeedeh Shojaee, Mohammadali Torbati, Adel Mirza Alizadeh, Matin Alizadeh Pages 335-347
    Antibiotic therapy is among the most important treatments against infectious diseases andhas tremendously improved effects on public health. Nowadays, development in using thistreatment has led us to the emergence and enhancement of drug-resistant pathogens which canresult in some problems including treatment failure, increased mortality as well as treatmentcosts, reduced infection control efficiency, and spread of resistant pathogens from hospitalto community. Therefore, many researches have tried to find new alternative approaches tocontrol and prevent this problem. This study, has been revealed some possible and effectiveapproaches such as using farming practice, natural antibiotics, nano-antibiotics, lactic acidbacteria, bacteriocin, cyclopeptid, bacteriophage, synthetic biology and predatory bacteriaas alternatives for traditional antibiotics to prevent or reduce the emergence of drug resistantbacteria.
    Keywords: Drug resistance, Food borne pathogens, Food safety, Health, Prevention strategies
  • Shreya Shanbhag, Akshatha Nayak, Reema Narayan, Usha Yogendra Nayak * Pages 348-359
    Skin, being one of the vital organs and a protective barrier needs to be pampered and taken careof from early childhood. It is the most visible and the widest exposed organ and by far reflectsthe general health condition and the aging process in humans. Both intrinsic and extrinsicfactors contribute to this complex biological process of skin aging. In recent times, skin healthand its beauty is perceived as an indicator of one’s health which has resulted in an increasingdemand for anti-aging products. Exposure to UV radiation is considered to be one of the factorsresponsible for aging termed as photoaging. In this review, we have discussed the various factorswhich may accelerate the process of skin aging. Various approaches and strategies to delay theprocess of skin aging have been emphasized upon. The patents filed in the area of anti-agingand sunscreen products have also been reviewed to gain an insight into the new formulationswhich have been developed as an anti-aging product. There has been a tremendous rise in thecosmetic and cosmeceuticals market with products having a dual activity of anti-aging and sunprotection. Research is constantly on the rise to ensure the safety of these products. Alternativesto the current topical application of sunscreen are being considered to overcome the drawbackof reapplication of the sunscreen often which can be a boon to the cosmeceutical market.
    Keywords: Aging, Anti-aging, Nanoparticles, Photoprotection, Sunscreen, Sun protection factor (SPF)
  • Ali Alirezaie, Mohamd Barati*, Meisam Barati, Hussein Abbasi Pages 360-373
    Cancer is an abnormal cell growth which tends to proliferate in an uncontrolled way and, insome cases, leads to metastasis. If cancer is left untreated, it can immediately cause death. Theuse of magnetic nanoparticles (MNPs) as a drug delivery system will enable drugs to targettissues and cell types precisely. This study describes usual strategies and consideration for thesynthesis of MNPs and incorporates payload drug on MNPs. They have advantages such asvisual targeting and delivering which will be discussed in this review. In addition, we consideredbody magnetic field to make drug delivery process more effective and safer by the applicationof MNPs and tumor-on-chip.
    Keywords: Body magnetic fields, Cancer, Drug delivery, Magnetic nanoparticles
  • Hridya Jayamohananan, Maneesha Kalappurackal, Aneesh T P * Pages 374-381
    Neurological and psychiatric disorders occur in about 6 percent of the global populationindicating a significant amount of people suffering from neurological disorder on a varying rangein day to day life. On an extensive view, there is a critical requirement for the development ofan alternative biomarker for these conditions. The thwart found in developing a biomarker isthe difficulty in identifying a serum biomarker as these are mostly limited to the central nervoussystem (CNS). Serotonin being a neurotransmitter synthesized in the raphe nuclei of the braincould serve as an alternative biomarker. Here, the limitation is that it’s quickly metabolizedby the mitochondrial enzyme MAO to 5-hydroxy indole acetic acid (5HIAA). This subsequentmetabolite can be used for the analysis of serotonin levels in brain by analysing its concentrationin the cerebrospinal fluid (CSF). Many theories suggest that the variations in serotonin levelcould lead to the development of many neurological and psychiatric disorders like Alzheimer’sdisease (AD), schizophrenia, depression and so on. A decreased level is noticed in these patientsbut this could either be due to decreased production or increased reuptake of serotonin fromthe neuronal synapses. For instance, we know that a patient with depression shows a significantreduction in the levels of 5HIAA, due to the location of the raphe nuclei within regions ofmemory and cognition. Similarly, it does shows variation in AD and mild cognitive disorder.Evolving of 5HIAA as a biomarker, could be more delicate and enhanced strategy for monitoringthese disorders.
    Keywords: Serotonin, Tryptophan, Raphe nuclei, 5-hydroxyindole acetic acid, Neurological functions
  • Teuku Nanda Saifullah Sulaiman, Dwi Larasati, Akhmad Kharis Nugroho, Syaiful Choiri * Pages 382-392
    Purpose
    To assess the effect of the lactic acid (LA)-to-glycolic acid (GA) molar ratio andpolyethylene glycol (PEG) concentration on the formation of poly-lactide co-glycolide acid(PLGA)-PEG-PLGA co-block polymers simultaneously using statistical approach.
    Methods
    A 22 full factorial design with the addition of a point in the center of the design, namelycurvature, was applied. Fourier transform infrared (FTIR), differential scanning calorimetry(DSC), and nuclear magnetic resonance (NMR) were performed to confirm the formation of theco-block polymer. Simvastatin (SMV), a drug model was incorporated into the nano-polymericmicellar (NpM) of PLGA-PEG-PLGA followed by solubility phase, particle size, zeta potential,and entrapment efficiency characterizations.
    Results
    FTIR, DSC, and NMR successfully confirmed the formation of co-block polymers.Solubility of SMV increased from 2 to 44-folds depending on co-block concentration withentrapment efficiency of 59%-80%. The NpM had size in the range of 206 to 402 nm withnegative zeta potential. LA to GA ratio had greater effect on particle size reduction and increasingof co-polymer length. In addition, it had higher contributions on increasing of solubility andentrapment efficiency of SMV than PEG.
    Conclusion
    According to these findings, the LA to GA ratio and PEG concentration gained agreat consideration in order to prepare the PLGA-PEG-PLGA co-block which fulfilled the qualitytarget product profile of NpM delivery system.
    Keywords: PLGA-PEG-PLGA, Co-block polymer, Nano polymeric micellar, Thermo- sensitive polymer
  • Yaser Shahparast, Masoud Eskandani, Ahmad Rajaei, Ahmad Yari * Pages 393-400
    Purpose
    This study aimed to improve the pharmacokinetic behavior of polyunsaturatedfatty acids (PUFAs) oxidation to enhance oxidative stability for inhibiting formation of toxichydroperoxides, develops off-flavors and shortens shelf-life.
    Methods
    Nanostructured lipid carrier (NLC) co-encapsulating omega-3 fish oil and α-tocopherolwas successfully prepared by melt blending and hot sonication method to enhance the oxidativestability of the fish oil. Encapsulation efficiency (EE) and in vitro release, the oxidative stabilityof prepared nanoparticles (NPs) were measured using detection of peroxide value (PV) andthiobarbituric acid (TBA) during 40 days.
    Results
    Electron microscopy and particle size analysis showed dispersed and homogenousNPs with an average diameter of 119 nm. Sustained oil release at a physiologic pH, and longtermstability in terms of the size, zeta, and dispersity of NPs was achieved after 75 days ofstorage. The omega-3 fish oil co-encapsulated with α-tocopherol in the NLC possessed betteroxidative stability compared with the all other formulations. Also, it was found that the NLC asan encapsulation method was more successful to inhibit the formation of the primary oxidationproducts than the secondary oxidation products.
    Conclusion
    Generally, these findings indicated that co-encapsulation of fish oil and α-tocopherolwithin the NLC can be a suitable delivery system in order to enrich foodstuffs, in particular clearbeverages.
    Keywords: Co-encapsulation, Omega-3 fish oil, Oxidative stability, Nanostructured Lipid Carriers, α -tocopherol
  • Atinderpal Kaur, Reema Gabrani, Shweta Dang * Pages 401-408
    Purpose
    Nanoemulsions (NEs) of polyphenon 60 (P60) and cranberry (NE I) and P60 andcurcumin (NE II) were prepared with the aim to enhance anti-bacterial potential and tounderstand the mechanism of anti-bacterial action of the encapsulated compounds.
    Methods
    To evaluate the antibacterial potential of the developed NE, microtiter biofilmformation assay was performed. The cytotoxicity analysis was done to assess the toxicity profileof the NEs. Further antibacterial analysis against uropathogenic strains was performed to checkthe developed NEs were effective against these strains.
    Results
    In microtiter dish biofilm formation assay, both NE formulations inhibited the growthmore effectively (Av. % inhibition ~84%) as compared to corresponding aqueous solution(Av. % inhibition ~64%) and placebo (Av. % inhibition ~59%) at their respective minimuminhibitory concentration (MIC) values. Cytotoxicity analysis using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay) showed that the formulations were nontoxic toVero cells. The antibacterial studies against uropathogenic resistant strains also showed that NEseffectively inhibited the growth of bacterial strains.
    Conclusion
    From different studies it was concluded that both the NE’s were able to inhibitbacterial strains and could be further used for the treatment of urinary tract infection (UTI). Theantibacterial activity of developed NEs showed that these could be used as alternative therapiesfor the treatment of UTI.
    Keywords: Biofilm, Cranberry, Curcumin, Cytotoxicity, Green tea, Nanoemulsion
  • Jalil Rashedi, Amir Ghorbani *, Mehran Mesgari, Ali Dastranj, Shadi Yaqoubi, Davoud Sanajou, Zahra Ashrafi, Ali Namvaran, Ali Mohammadi, Jalal Mohammadi, Behzad Baradaran * Pages 409-415
    Purpose
    This study was aimed to evaluate the site-specific drug delivery of 5-FU with chitosan(CS) as a carrier and quercetin (Qu) against induced colon cancer in Wistar rats.
    Methods
    Cross-linked CS-Qu nanoparticles (NPs) were prepared by ionotropic gelation method.Physicochemical characterization of NPs was performed by Fourier-transform infrared (FTIR)spectroscopy, dynamic light scattering (DLS), in vitro drug release, and drug loading efficiency(LE). 1, 2-Dimethylhydrazine (DMH) and dextran sulfate sodium (DSS) were applied to induceadenocarcinoma tumors on inbred male Wistar rats’ colon. The treatment group of rats wasadministered through enema with NPs dispersion. Hematoxylin and eosin staining were performedto the histopathological examination of tumors.
    Results
    Zeta potential and particle size for NPs were +53.5 ± 5 mV and 179 ± 28 nm, respectively.About 96% Qu LE was obtained with a maximum release of 5.63 ±1.59% and 4.62 ± 1.33%after 24 hours in PB solution with pH values of 6 and 7.4, respectively. The numbers of 8 to 21tumors were observed in all rats administered with DMH and DSS. Significantly decreasing ofmicrovascular density and mitosis count was detected in the treatment group in comparison withcancerous group (P = 0.032 for the former compared to P = 0.016 for the later), respectively.Furthermore, the treatment group showed a high apoptosis rate (P = 0.038).
    Conclusion
    The developed Qu-loaded CS NPs were good candidates for site-specific andsustained drug release in enema treatment. Decreasing of microvascular density and mitosiscount, along with increasing the apoptosis percent in the treatment group proved that the NPscould have promising results in site-specific and sustained drug delivery against colorectal cancer.
    Keywords: Anti-tumor, Chitosan, Colon, Nanoparticles, Quercetin, Rats, Wistar
  • Rafael Souza, Douglas Vieira *, Luane Ferreira Garcia , Eric de Souza Gil ORCiD, Vernon Sydwill, Flavio Marques Pages 416-422
    Purpose
    Jenipapo fruit (Genipa americana L) is a natural source of polyphenol oxidases (PPOs)whose potential in pharmaceutical analysis is noteworthy. Henceforth, this work reports theelectrochemical study of a low-cost PPO-based biosensor produced from the crude extract ofJenipapo fruits and accounts a practical approach to employ this biosensor in the determinationof methyldopa and paracetamol in pharmaceutical samples.
    Methods
    In order to investigate the electrochemical properties of the biosensor, theoreticaland practical approaches were employed, and both samples and the biosensor were analyzedthrough electrochemical impedance spectroscopy (EIS) and voltammetric techniques, namely:differential pulse voltammetry (DPV) and cyclic voltammetry (CV).
    Results
    showcased that the biosensor presented good analytical features, as well as lowdetection limits (8 μmol L-1 for methyldopa and 5 μmol L-1 for paracetamol). The relativestandard deviation was less than 5% mid-assay.
    Conclusion
    The use of this biosensor is a reliable, low cost and useful alternative in thepharmaceutic determination of phenolic drugs (e.g. methyldopa and paracetamol).
    Keywords: Jenipapo (Genipa americana L.), Biosensor, Methyldopa, Paracetamol
  • Mahdi Barazesh, Zohreh Mostafavipour, Soudabeh Kavousipour, Shiva Mohammadi, Pooneh Mokarram * Pages 423-431
    Purpose
    GnRH-DFF40 (gonadotropin releasing hormone - DNA fragmentation factor 40) isa humanized recombinant immunotoxin and serves as a prospective candidate for targetedtherapy of gonadotropin releasing hormone receptor (GnRHR) overexpressing malignancies.However, its production in Escherichia coli in a soluble and functional form still remains achallenge. Here we introduce two successful and reproducible conditions for production andpurification of "difficult-to-express" GnRH-DFF40 protein.
    Methods
    A synthetic codon optimized GnRH-DFF40 fusion gene was cloned in pET28aplasmid. Two methods including high cell density IPTG induction (HCDI) and autoinductionmethod (AIM) with a focus on obtaining high cell density have been investigated to enhance theprotein production in (E. coli). Moreover, to obtain higher protein production several factors inthe AIM method including carbon sources, incubation time and temperature, plasmid stabilityand double colony selection, were optimized.
    Results
    Remarkable amounts of soluble GnRH-DFF40 protein were achieved by both methods.Cell density and protein yields in AIM was about 1.5 fold higher than that what obtained usingHCDI. Initial screening showed that 25ºC is better to achieve higher protein production in bothmethods. pH alterations in AIM were maintained in a more constant level at 25ºC and 37ºCtemperatures without any detrimental effects on cell growth during protein production phaseup to 21 hours after incubation. Plasmid stability during growth and expression induction phasewas maintained at a high level of 98% and 96% for AIM and HCDI methods, respectively. Afterparameter optimization and double colony selection in AIM, a very high yield of recombinantprotein was achieved (528.3 mg/L).
    Conclusion
    With the optimization of these high cell density expression methods, reproduciblemanifold enhancement of soluble protein yields can be achieved for "difficult-to-express"GnRH-DFF40 compared to conventional expression methods.
    Keywords: Humanized recombinant immunotoxin, GnRH-DFF40 chimeric protein, Autoinduction method (AIM), High cell density IPTG induction (HCDI), Targeted therapy
  • Valcilaine Teixeira, Janaína Barros de Menezes, Josué Carinhanha, Maria Lysete de Assis, João Xavier de Araújo, Ticiano Gomes, Irinaldo Diniz, Luciano Aparecido, Camila Braga Dornelas * Pages 432-438
    Purpose
    Quercetin is a flavonoid known for its therapeutic properties and for formingcomplexes. Although the antimony-quercetin (SbQ) complex has been produced before, noprevious exploration of its characteristics has been published in literature. Thus, this study aimedto characterize this complex, assess its stability and investigate its complexation site through itsantibacterial activity.
    Methods
    The SbQ complex was synthetized using Sb(III) potassium tartrate trihydrate andquercetin anhydrous (1:1) (v/v) as a solution and dried using three
    methods
    rotaevaporation,lyophilization and spray drying. The material, in solution, was analyzed by UV-vis andfluorimetry; and, in the powder, by X-ray diffraction (XRD), both scanning electronic andfluorescence microscopy and infrared spectroscopy (FT-IR). Antimicrobial activity was evaluatedvia broth microdilution.
    Results
    UV-vis exhibited a shoulder peak at 291 nm indicating metal chelation at C-ring ofquercetin and confirmed 1:1 stoichiometry. Spectrofluorimetry showed an increase of intensitywith the complex formation with an emission band (525 nm). After drying, XRD and SEMindicated loss of crystallinity and a difference in shape and size of the complex compared to itsprecursors. FT-IR suggested by a shift of frequency of the carbonyl group (1661 cm-1) that thequercetin bond to antimony by the C-3, followed by positions C-5 and C-4 carbonyl, which hasbeen confirmed by MIC through the structure-activity relationship of the antibacterial activityof quercetin.
    Conclusion
    These results provided a characterization of SbQ complex with the confirmation ofits binding site, working as a guide for future studies involving this complex.
    Keywords: Antimony, Binding sites, Flavonoids, Quercetin
  • Mahshid Mohammadian, Shima Zeynali, Mohammad Hassan Khadem, Yousef Rasmi, Anahita Fathi, Fatemeh Kheradmand * Pages 439-444
    Purpose
    Dihydropyrimidine dehydrogenase (DPD) is the principal enzyme in the catabolismof fluoropyrimidine drugs including capecitabine. A recent report has suggested that oxaliplatinchemotherapy is associated with elevated DPD levels and chemoresistance pattern. As a newlydeveloped chemotherapeutic agent, 17-allyloamino-17-demethoxy-geldanamycin (17-AAG)can be effective in combination therapy with oxaliplatin and capecitabine in colorectal cancer(CRC). DPD expression level can be a predictive factor in oxaliplatin and capecitabine-basedchemotherapy. We evaluated DPD in mRNA and protein levels with new treatments: 17-AAG incombination with oxaliplatin and capecitabine in HT-29 and HCT-116 cell lines.
    Methods
    Drug sensitivity was determined by the water-soluble tetrazolium-1 assay in aprevious survey. Then, we evaluated the expression levels of DPD and its relationship with thechemotherapy response in capecitabine, oxaliplatin, and 17-AAG treated cases in single andcombination cases in two panels of CRC cell lines. DPD gene and protein expression levels weredetermined by real-time polymerase chain reaction and western blotting assay, respectively.
    Results
    DPD gene expression levels insignificantly increased in single-treated cases versusuntreated controls in both cell lines versus controls. Then, the capecitabine and oxaliplatinwere added in double combinations, where DPD gene and protein expression increased incombination cases compared to pre-chemotherapy and single drug treatments.
    Conclusion
    The elevated levels of cytotoxicity in more effective combinations could be relatedto a different mechanism apart from DPD mediating effects or high DPD level in the remainingresistance cells (drug-insensitive cells), which should be investigated in subsequent studies.
    Keywords: Colorectal cancer, Oxaliplatin, Capecitabine, 17-AAG, Dihydropyrimidine dehydrogenase
  • Edy Meiyanto*, Herwandhani Putri, Yonika Arum, Rohmad Yudi, Riris Istighfari, Muthi Ikawati, Beni Lestari, Noriko Yoneda, Ikuko Nakamae, Masashi Kawaichi, Jun, Ya Kato Pages 445-452
    Purpose
    Pentagamavunon-1 (PGV-1) is a curcumin analogue that shows cytotoxic activity invarious cancer cells. In this study, we evaluated the effect of PGV-1 on a highly metastatic breastcancer cell line, the 4T1 cells, as an anti-metastatic and anti-proliferative agent.
    Methods
    Cell viability was evaluated using MTT assay; while cell cycle profile, apoptosisincidence, and ROS intracellular level were determined by flow cytometry. Cell senescence wasobserved under senescence-associated-β-galactosidase (SA-β-gal) staining assay. The expressionof matrixmetalloproteinase-9 (MMP-9) was determined using immunoreaction based-ELISA,while other proteins expression were detected using immunoblotting.
    Results
    Curcumin and PGV-1 showed cytotoxic effects on 4T1 cells with IC50 value of 50 and4 μM, respectively. The cytotoxic activity of PGV-1 was correlated to the induction of G2/M cellcycle arrest and cell senescence. Furthermore, PGV-1 increased the accumulation of intracellularROS level. We also revealed that PGV-1 bound to several ROS-metabolizing enzymes,including glyoxalase I (GLO1), peroxiredoxin 1 (PRDX1), N-ribosyldihydronicotinamide:quinone reductase 2 (NQO2), aldo-keto reductase family 1 member c1 (AKR1C1). As an antimetastaticagent, PGV-1 showed less inhibitory effect on cell migration compared to curcumin.However, PGV-1 significantly decreased MMP-9 protein expression in a dose-dependentmanner suggesting it still potent to inhibit metastatic cells.
    Conclusion
    Overall, our findings suggest that PGV-1 is potential to be developed as an antiproliferativeand anti-metastatic agent.
    Keywords: Anti-proliferative, Metastatic inhibitor, PGV-1 (curcumin analogoe), Reactive oxygen species (ROS), The 4T1 cells
  • Riris Istighfari, Nur Dina Amalina, Gagas Pradani, Rohmad Yudi, Muthi Ikawati, Annisa Khumaira, Jun, Ya Kato, Edy Meiyanto * Pages 453-461
    Purpose
    Genistein, a soy isoflavone, exhibits a biphasic effect on cells proliferation withsome different effects between ER-alpha and ER-beta. The objective of this present study is todetermine the modulatory effect based on cell cycle progression under genistein treatment incombination with 17-β estradiol (E2) on CHO-K1 cells.
    Methods
    The effect of genistein 0.1-100 μM on cells proliferation was examined by MTT assay.The modulation of genistein and estradiol (E2) on cell cycle and apoptosis were observed byusing flowcytometry with PI and PI/AnnexinV staining, respectively. Moreover, the effect ofgenistein and E2 on senescence cells, and ROS level were determined by senescence-associatedβ-galactosidase (SA β-gal) staining and by using flowcytometry with 2’, 7’–dichlorofluorescindiacetate (DCFDA) staining, respectively. The expression level of the cell cycle and senescenceprotein markers were observed by immunoblotting.
    Results
    Single treatment of genistein at physiologically achievable (low) concentration (<2 μM)induced proliferation of CHO-K1 cells while at a pharmacological (high) concentration (50 and100 μM) suppressed cells proliferation. Interestingly, treatment of genistein at the physiologicalconcentration in combination with E2 for 24, 48 and 72 h decreased cells viability on CHO-K1cells compared to untreated cells. Further analysis of the cells showed that 50 μM genisteininduced G2/M phase accumulation and induced apoptosis. Moreover, genistein induced cellsenescence and increased ROS level. Immunoblotting analysis showed the decreasing of ERalpha,Bcl2, and ppRb protein level upon treatment of 1 μM Gen and 1 nM E2.
    Conclusion
    Our results suggest that the cell proliferation inhibitory mechanism of genistein atpharmacological concentration involved the induction of cell senescence, and the elevationof ROS level. Moreover, the decreased of cells proliferation upon treatment of physiologicalconcentration of genistein in combination with E2 may be correlated with the alteration of ERexpression.
    Keywords: Genistein, Cell cycle, Senescence, Apoptosis, Reactive oxygen species (ROS)
  • Atefeh Satari, Sayed Asadollah Amini, Elham Raeisi, Yves Lemoigne, Esfandiar Heidarian* Pages 462-469
    Purpose
    Prostate cancer is as far the most prevalent male cancer. Rutin (a glycoside fromquercetin flavonoid) displays antioxidant activity leading to cell apoptosis. Combined effects ofrutin with the widely used anti-cancer drug, 5-fluorouracil (5-FU), on prostate cancer cell line(PC3) was investigated herein.
    Methods
    Different concentrations of combined 5-FU and rutin were applied to PC3 cellscompared to separate treatment for 48 hours. Cell viability, as well p53 gene expressionrespectively were assessed by MTT assay and real-time quantitative polymerase chain reaction(qPCR). Changes of Bcl-2 signal protein and apoptosis were determined using western blotand flow cytometry procedures, respectively. Clonogenic assay was used to colony countsassessment.
    Results
    50% inhibitory concentration (IC50) of separate cell treatment with either rutin and5-FU respectively were 900 μM and 3Mm, while combination index (CI) of combined 5-FU/rutin application reached a level of synergistic effects (0.33). Combination of 5-FU/rutinenhanced apoptosis and p53 gene expression in PC3 cells. PC3 cell colony counts and Bcl-2signaling protein were decreased by 5-FU/rutin combination.
    Conclusion
    Synergistic effects of 5-FU/rutin combination on PC3 cells line enhanced apoptosis,p53 gene expression, and down-regulation of Bcl-2 protein, compared to control separateapplication. 5-FU/rutin combination does seem an interesting therapeutic pathway to be furtherinvestigated.
    Keywords: Apoptosis, Rutin, 5-Fluorouracil, Prostate Cancer
  • Elahe Naderali, Behnaz Valipour, Amir Afshin Khaki, Jafar Soleymani Rad, Alireza Alihemmati, Mohammad Rahmati, Hojjatollah Nozad * Pages 470-480
    Purpose
    The PI3K/Akt signaling pathway regulates cell growth, proliferation and viability inhematopoietic cells. This pathway always dysregulates in acute lymphoblastic leukemia (ALL).PTEN and P53 are tumor suppressor genes correlated with PI3K/Akt signaling pathway, and bothhave a tight link in regulation of cell proliferation and cell death. In this study, we investigatedthe effects of dual targeting of PI3K/Akt pathway by combined inhibition with nvp-BKM-120(PI3K inhibitor) and MK-2206 (Akt inhibitor) in relation with PTEN and P53 on apoptosis andproliferation of leukemia cells.
    Methods
    Both T and B ALL cell lines were treated with both inhibitors alone or in combinationwith each other, and induction of apoptosis and inhibition of proliferation were evaluated byflow cytometry. Expression levels of PTEN as well as p53 mRNA and protein were measured byreal-time qRT-PCR and western blot, respectively.
    Results
    We indicated that both inhibitors (BKM-120 and MK-2206) decreased cell viability andincreased cytotoxicity in leukemia cells. Reduction in Akt phosphorylation increased PTEN andp53 mRNA and p53 protein level (in PTEN positive versus PTEN negative cell lines). Additionally,both inhibitors, particularly in combination with each other, increased apoptosis (evaluatedwith Annexin V and caspase 3) and reduced proliferation (Ki67 expression) in leukemia cells.However, administration of IL7 downregulated PTEN and P53 mRNA expression and rescuedcancer cells following inhibition of BKM-120 and MK-2206.
    Conclusion
    This investigation suggested that inhibition of Akt and PI3K could be helpful inleukemia treatment.
    Keywords: PI3K, Akt signaling, MK-2206, BKM-120, ALL, PTEN, P53
  • Masoud Modarresi, Marziyeh Hajialyani, Narges Moasefi, Farahnaz Ahmadi, Leila Hosseinzadeh * Pages 481-489
    Purposes
    In the present study, we tried for the first time to examine the anti-proliferative andanti-apoptogenic effect of Glabridin (Glab) toward three groups of cancer cells (SKNMC,H1299, and A2780). Furthermore, the possibility of co-administration of Glab with doxorubicin(DOX) to these cells was also examined to find out whether Glab can potentiate the cytotoxiceffect of this chemotherapy agent.
    Methods
    Different cellular assays (MTT, caspase-3 activity, MMP, RT-PCR analysis) were carriedout on the cancer cells treated with Glab.
    Results
    Cellular toxicity assay revealed that Glab can potentially reduce the viability of thesecells with IC50 concentrations up to 10, 12, and 38 μM toward A2780, SKNMC, and H1299 celllines, respectively. The results of MMP and caspase-3 activity assays, in association with theresults corresponding to the BAX and Bcl-2 gene expressions, altogether revealed that Glab canexert apoptogenic effect on these cells. The intrinsic mitochondrial pathway was found to bethe main mechanism, in which Glab induced apoptosis toward H1299 cells and SKNMC cells,while the apoptosis mechanism for A2780 cells could be probably through extrinsic pathway.Glab also potentiated the cytotoxic effect of DOX and its accumulation in H1299 cell line.
    Conclusion
    The results of this study revealed the promising cytotoxic role of Glab on differentcarcinoma cells. These data also suggested that co-chemotherapy method using Glab could beeffective for treatment of cancer, but further in-vivo and clinical studies are still needed to assurethese results.
    Keywords: Apoptosis, Cytotoxicity, Doxorubicin, Glabridin, Herbal medicine, Licorice
  • Javad Saffari, Chaleshtori, Ehsan Heidari, Fahimeh Moradi, Esfandiar Heidarian* Pages 490-496
    Purpose
    Since active plant ingredients can induce apoptosis in many tumors, in this studywe evaluate the apoptotic effects of thymoquinone (TQ) on PC3 cells. Also, we predicted theinteraction of TQ with BCL-XL, BCL-2, and MCL-1anti-apoptotic factors by computer-simulatedenvironment.
    Methods
    PC3 cells were treated with different concentrations of TQ (0- 80 μM) and IC50 wasdetermined using 3-(4, 5-dimethylthiaztol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay.Apoptotic and cytotoxicity effects of TQ were analyzed using flowcytometry and comet assay,respectively. Changes in energy and the molecular interactions of TQ with BCL-XL, BCL-2 andMCL-1 anti-apoptotic factors were investigated using simulation.
    Results
    IC50 value was 40 μM. TQ led to the destruction of the genome of PC3 cells and inducingapoptosis. Molecular dynamics (MD) revealed that the root mean-square deviation (RMSD),root mean square fluctuation (RMSF), radius of gyration (Rg), and the number of hydrogen andhydrophobic bonds between TQ and residues of BCL-2, BCL-XL and MCL-1were significantly(P < 0.001) changed. TQ makes a more stable and stronger connection with BCL-XL comparedto BCL-2 and MCL-1 and inhibits BCL-XL non-competitively.
    Conclusion
    Our results demonstrated that TQ not only led to apoptosis, at least partly, due toreduction in the Coil, Turn, and Bend structure of BCL-XL but also caused a decrease in the Rgand RMSD value of BCL-XL, MCL-1, and BCL-2.
    Keywords: Thymoquinone, Apoptosis, Cancer, Simulation
  • Homa Salami, Seyed Javad Mowal, Rasoul Moukhah, Zahra Hajebrahimi, Seyed Abdolhakim, Houri Edalat * Pages 497-504
    Purpose
    The histone deacetylases (HDAC) inhibitor, valproic acid (VPA), is a commonantiepileptic drug and is attractive for its broad range of therapeutic effects on many diseases. Ithas been employed as an inducer of pluripotency in some cultured cells. Conversely, VPA hasalso been employed as an inducer of in vitro differentiation in many other cells. Therefore, weemployed WJMSCs as a cellular target to evaluate the differential effects of of VPA on potencystate and differentiation level of Wharton’s Jelly mesenchymal stem cells (WJMSCs) in variousconcentrations and different culture mediums.
    Methods
    The isolated WJMSCs were cultured in DMEM (MSC medium). According to previousprotocols, WJMSCs were treated with 0, 0.5 and 1 mM VPA in MSC or embryonic stem cell (ESC)medium and 2 mM VPA in neural differentiation medium. Real-time polymerase chain reaction(PCR) and western blot analysis were performed for evaluating the expression of pluripotencymarkers. MTT and caspase assays were also performed on VPA-treated cells.
    Results
    The expression of pluripotency markers and the viability of the WJMSCs – determinedby MTT assay – were significantly increased after 0.5 mM VPA treatment in ESC medium. A 2mM VPA treatment in neural differentiation medium significantly diminished the expression ofpluripotency markers and the viability of WJMSCs.
    Conclusion
    According to our results, both VPA concentration and the medium context caninfluence VPA effects on WJMSCs. The differential effects of VPA on WJMSCs can reflect its widerange of effects in the treatment of various diseases.
    Keywords: Valproic acid, Wharton’s jelly mesenchymal stem cells, Potency, Neural differentiation
  • Mahdieh Ahmadiany, Mahshid Alavi, Zahra Hashemi, Mohammad Amin Moosavi, Marveh Rahmati Pages 505-509
    Purpose
    Despite recent advances in the diagnosis and treatment of rheumatoid arthritis (RA), thisinflammatory disease remains a challenge to patients and physicians. Recent evidence highlightsthe contribution of endoplasmic reticulum (ER) stress in the pathogenesis and treatment of RA.Herein, we study the expression of the ER stress sensor inositol-requiring enzyme 1α (IRE1α),as well as XBP1 splicing and the regulated IRE1-dependent decay (RIDD), in peripheral bloodmononuclear cells (PBMCs) from patients with RA compared with healthy controls.
    Methods
    The PBMCs from blood samples of RA patients and healthy volunteers were isolatedby a density gradient centrifugation method using Ficoll. The gene expression levels of GRP78/Bip, IRE1, XBP1s, micro-RNAs (miRNAs) were evaluated by real-time PCR.
    Results
    The expression of GRP78, IRE1, and XBP1s were increased in PBMCs of RA patientscompared with healthy controls. We further show that the RIDD targets (miRNA-17, -34a, -96,and -125b) were downregulated in RA samples.
    Conclusion
    This study can expand our knowledge on the importance of RNase activity ofIRE1α in RA and may offer new potentials for developing novel diagnostic and/or therapeuticbiomarkers.
    Keywords: Endoplasmic reticulum stress, Inositol-requiring enzyme 1, IRE1-dependent decay, microRNA, Rheumatoid arthritis