Iranian Journal of Basic Medical Sciences
Volume:23 Issue: 12, Dec 2020

Folate is an important water-soluble vitamin that is presented naturally in foods in particular vegetables, fruits, and whole grains. To which extent is this vitamin needed in our daily regimen is not fully known. Several studies have indicated that many complications, such as megaloblastic anemia, cardiovascular disease, neural tube defects, and numerous cancers, occur in humans when the body becomes deficient in folic acid. On the other hand, a few studies have shown thier concerns regarding the supplementation of folic acid, resulting in the development of existing tumors and alteration of normal patterns of DNA methylation. Although there is no clear evidence of aberrant DNA methylation and gene expression changes in response to “high” levels of folate  or folic acid intake, there are still some concerns. Therefore, its adverse effects especially on fetus and later stages of life should be carefully investigated.

Withania somnifera L. is a multipurpose medicinal plant of family Solanaceae occurring abundantly in sub-tropical regions of the world. The folk healers used the plant to treat several diseases such as fever, cancer, asthma, diabetes, ulcer, hepatitis, eyesores, arthritis, heart problems, and hemorrhoids. The plant is famous for the anti-cancerous activity, low back pain treatment, and muscle strengthening, which may be attributed to the withanolide alkaloids. W. somnifera is also rich in numerous valued secondary metabolites such as steroids, alkaloids, flavonoids, phenolics, saponins, and glycosides. A wide range of preclinical trials such as cardioprotective, anticancer, antioxidant, antibacterial, antifungal, anti-inflammatory, hepatoprotective, anti-depressant, and hypoglycemic have been attributed to various parts of the plant. Different parts of the plant have also been evaluated for the clinical trials such as male infertility, obsessive-compulsive disorder, antianxiety, bone and muscle strengthening potential, hypolipidemic, and antidiabetic. This review focuses on folk medicinal uses, phytochemistry, pharmacological, and nutrapharmaceutical potential of the versatile plant.

Microtubules have key roles in essential cellular processes such as mitosis, cell motion, and intracellular organelle transport. Increasing interest has been given to tubulin binding compounds after the introduction of taxanes into clinical oncology. The object of this study was  synthesis and biological evaluation of novel 5,6,7-trimethoxy quinolines as tubulin inhibitors. The cytotoxicity of the newly synthesized compounds was assessed against different human cancer cell lines including MCF-7, A2780, MCF-7/MX, A2780/RCIS, and normal cells. Compounds demonstrating the most antiproliferative activity, were chosen to examine their tubulin inhibition activity and their ability to arrest the cell cycle and induce apoptosis. Molecular docking studies and molecular dynamics simulation of compound 7e in the catalytic site of tubulin were performed. Most of the synthesized quinolines showed moderate to significant cytotoxic activity against human cancer cells. Compounds 7e and 7f, possessing N-(4-benzoyl phenyl) and N-(4-phenoxy phenyl), respectively, exhibited the most antiproliferative activity more potent than the other compounds and exhibited similar antiproliferative activity on both resistant and parental cancer cells. Flow cytometry analysis of A2780, A2780/RCIS, MCF-7, and MCF-7/MX cancer cells treated with 7e and 7f exhibited that these compounds arrested the cell cycle (at the G2/M phase) and induced cellular apoptosis in A2780 cancer cells. These quinolines inhibited tubulin polymerization in a way resembling that of CA-4. Molecular dynamics simulation and molecular docking studies of compound 7e into the binding site of tubulin displayed the probable interactions of 7e with the binding site of tubulin.

Immune responses are tightly regulated in the development of clonorchiasis. However, the adaptive immune regulatory pathway contributing to the pathological processes of clonorchis sinensis (C. sinensis) infection is still not clear. In this study, we assessed the dynamic changes of CD4+T cell subsets and the related cytokines as well as transcription factors during C. sinensis infection. We used female FVB mice to establish the infection model. H&E and Masson’s stain were performed in 2 or 8 weeks post-infection (PI) liver of C. sinensis. The percentages of splenic Th1, Th2, and Treg in CD4+T cells were detected by flow cytometry. The transcription factors T-bet, GATA3, Foxp3, and RORγt gene expression were detected by qPCR. The protein expression of IL-10, IL-17, IL-4, IL-2, and Tumor Necrosis Factor-α (TNF-α) were examined using ELISA. The percentages of splenic Th1, Th2, and Treg in CD4+T cells were significantly increased in both 2 and 8 weeks PI of C. sinensis, while the ratio of Treg/Th17 as well as the percentage of Treg in serum was gradually increased during the development of infection. The expressions of T-bet, GATA3, Foxp3, and RORγt were increased in 8 weeks PI. Serum levels of IL-10, IL-17, IL-4, and IL-2 were profoundly increased in infected mice, while the concentrations of TNF-α increased to peak two weeks PI. Our results suggested that the imbalance of CD4+T cell subsets may regulate and contribute to an appropriate compromise between pathology, tissue repair, and elimination in a susceptible murine host.

Celecoxib (CLX), a selective cyclooxygenase-II (COX-2) inhibitor, has been used for management of several inflammatory disorders. The present study aimed to explore the role of peroxisome proliferator-activated receptor-gamma (PPARγ) in CLX induced anti-inflammatory response in rats.

Carrageenan-induced paw edema was used as an acute inflammation model. Rats were treated with various intra-peritoneal (IP) doses of CLX (0.3–30 mg/kg) and pioglitazone (PGL; PPARγ agonist, 1–20 mg/kg) alone or in combination. Amounts of PPARγ, COX-2, and prostaglandin E2 (PGE2) in paw tissue, and extents of TNF-α and IL-10 in serum were measured. Moreover, levels of oxidative stress parameters as malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GPx) activity in the cortex, hippocampus, and paw tissues were also determined.  

CLX and PGL dose-dependent administration (IP), alone or in combination reduced carrageenan-induced paw edema. Further, both agents, alone or in combination, reduced either the amounts of COX-2, PGE2, and MDA in the inflamed paw, and the levels of TNF-α in serum which were elevated by carrageenan. Both drugs also increased both levels of PPARγ, GSH, GPx activity in paws, and serum levels of IL-10 that were decreased by carrageenan. Intraplantar injection of GW-9662 (IPL), a selective PPARγ antagonist, inhibited all biochemical modifications caused by both single and combined drug treatments.

CLX produced its anti-inflammatory effects probably through PPARγ receptor activation. Besides, increased anti-inflammatory effects of CLX with PGL suggest that their combination might be applied for the clinical management of inflammation especially in patients suffering from diabetes.

Escherichia coli is one of the most important causes of urinary tract infections (UTIs). The aim of this study was to determine antimicrobial resistance, resistance and virulence genes; phylogenetic groups and identify the epidemiologic features of uropathogenic E. coli (UPEC) isolates by multilocus sequence typing (MLST).

One hundred isolates of E. coli from inpatients with UTIs were collected in Kerman, Iran. Antimicrobial susceptibility testing, ESBLs, AmpC production and biofilm formation were performed by phenotypic methods. Phylogenetic groups, resistance and virulence genes were detected. Molecular typing of isolates was performed by MLST.

In this study, 76% of isolates were multidrug-resistant. The blaCTX-M-15 and blaTEM were the dominant ESBL-encoding gene. Among 63 ciprofloxacin-resistant isolates, the frequency of qnrS (15.8%), qnrB (9.5%), and aac (6’)-Ib (25% ) genes was shown. Fifty-five present of isolates were classified as week biofilm, (14%) moderate biofilm, and (5%) strong. The predominant phylogenetic group was B2 (3) .  The prevalence of virulence genes ranged fimH (93%), iutA (66%), KpsmtII (59%), sat (39%), cnf (28%) and hlyA (27%). According to MLST results, 14 sequence types (ST) including ST-693, ST-90, ST-101, ST-1664, ST-2083, ST-131, ST-4443, ST-744, ST-361, ST-405, ST-922, ST-648, ST-5717and ST-410 were detected, indicating a high degree of genotypic diversity.

We identified a high frequency of the ST131 clonal group among UTIs. These data show an important public health threat, and so further studies to control the dissemination and risk factors for acquisition of the ST131 clonal group and other STs are needed to make effective control.

This research was carried out to investigate the hypoglycemic activity of the ethyl acetate (EtOAc) extract from the roots of Smilax glabra Roxb, which strongly exhibit inhibitory activity against α-glucosidase and α-amylase on in vivo type 2 diabetic model. Column chromatography combined with crystallization was used to isolate the active fraction and compounds. Chemical structures of the compounds were determined based on the analysis of the spectroscopic data and comparison with the literature data. The α-glucosidase inhibitory activity (AGI) and the α-amylase inhibitory activity (AAI) were determined quantitatively spectrophotometrically using p-nitrophenyl α-D-glucopyranoside and soluble starch as substrates, respectively. The hypoglycemic activity was examined by evaluating its effects on glucose and insulin levels, insulin resistance, and histopathology of the pancreatic islets and livers in diabetic induced mice administrated with nicotinamide-streptozotocin. The EtOAc extract and the bioactive compounds astilbin and 5-O-caffeoylshikimic acid in the extract were isolated and confirmed in structures, AGI, and AAI. The treatment at the doses of 500 and 1000 µg/kg of body weight reduced blood glucose levels down to the physiological level of the physical controls in the diabetic mice after two weeks (p <0.05). Moreover, the treatment improved insulin sensitivity. Histopathology analysis showed recovering effects in the size of the pancreatic islets and no damaging effects on the liver after treatment compared with the control group. Our data suggest that the EtOAc extract possesses hypoglycemic activity and has an antidiabetic potential for therapeutic applications.

Ferula ovina is an Iranian medicinal plant. Tschimgine and stylosin are two of its major monoterpene derivatives. In this study, we proceeded to investigate some fungal endophytes from F. ovina that can produce plant secondary metabolites. The isolated endophytic fungi were fermented in potato dextrose broth (PDB) medium and their extracts were screened for the presence of the plant compounds by liquid chromatography-tandem mass spectrometry (LC-MS). Endophytes identification was performed by morphological and molecular methods. Three markers (ITS, LSU, and TEF1) were used for accurate molecular identification. Forty isolates from 9 different genera of endophytic fungi were identified, of which two recently reported species of O. ferulica and Pithoascus persicus were able to produce tschimgine and stylosin. These fungi can be used as a substitute for the production of plant’s medicinal compounds independent of wild populations of the source plant.

Evidence shows that sleep deprivation (SD) disrupts the formation of hippocampus-related memories. Moreover, α2 adrenergic receptors that are wildly expressed in the CA1 hippocampal region have a significant role in modulating both sleep and memory formation. In the present research, we wanted to investigate the effect of stimulation and blockage of CA1 α2 adrenergic receptors by clonidine (an agonist of α2 adrenergic receptor) and yohimbine (an antagonist of α2 adrenergic receptor), respectively, on memory retention impairment induced by REM SD (RSD) in rats.

Multiple platform apparatus were used to induce RSD, and the passive avoidance task was used to assess memory consolidation. Clonidine and yohimbine were injected intra-CA1.

The results showed that RSD (for 24 and 36, but not 12 hr) decreased memory retention, with no effect on locomotion. Post-training intra-CA1 infusion of a subthreshold dose of yohimbine (0.001 μg/rat) did not alter, while clonidine (0.1 μg/rat) restored memory retention impairment induced by RSD (24 and 36 hr). Also, none of the interventions did not influence locomotor activity.

Our data strongly showed that CA1 α2 adrenergic receptors have a critical role in RSD-induced memory retention impairment.

The human apolipoprotein E4 (APOE4) is associated with various brain injuries and neurodegenerative changes. Curcumin is an active ingredient isolated from the root of turmeric and is believed to have therapeutic effects on neurodegenerative diseases. The aim of this study was to investigate the effects of curcumin on APOE4-induced neurological damage and explore its molecular mechanisms. SH-SY5Y cells were pretreated with curcumin for 24 hr and transfected with human APOE4 gene using Lipofectamine 2000. Then, the effect of curcumin on the transfected cells was detected by ELISA, immunofluorescence staining and Western blot. The production or expression of proinflammatory cytokines and proteins, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), nitric oxide (NO), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) was significantly increased in SH-SY5Y cells transfected with APOE4, and curcumin inhibited APOE4-induced cellular inflammatory damage. Western blot analysis showed that, after transfection with APOE4, the expression of total nuclear factor kappa B (NF-κB) p65 and p-NF-κB p65 in the nucleus was increased, and curcumin inhibited the nuclear translocation of p65. The overexpression of APOE4 inhibited the expression of peroxisome proliferator-activated receptor-γ (PPARγ), whereas curcumin reversed and increased the expression of PPARγ protein. Down-regulating PPAR-γ with the inhibitor GW9662 and the shPPARγ gene confirmed that the NF-κB signaling pathway was inhibited by PPARγ. This study suggests that APOE4 overexpression can induce cellular inflammatory damage, and pretreatment of curcumin could exert an anti-inflammatory effect by upregulating the expression of PPARγ to inhibit the activation of NF-κB signaling pathway.

The modulatory effect of deep inspiration (DI) on airway constriction is impaired in asthma. However, mechanisms underlying this impairment are not clear. Since there is evidence indicating that Rho-kinase activation mediates force maintenance under oscillatory strain, we investigated the impact of Rho-kinase inhibition on the bronchodilatory effect of DI in ovalbumin (OVA) sensitized guinea pigs.

forty-eight male Dunkin Hartley guinea pigs were divided into 8 groups including saline/ constant, saline/DI, OVA/constant, OVA/DI, Rho-I/OVA/constant, Rho-I/OVA/DI, OVA-Rho-I/MCh/constant, and OVA-Rho-I/MCh/DI. Animals were subjected to 12 inhalations of OVA or saline aerosol. Guinea pigs in Rho-I/OVA/constant or DI groups were treated with the Rho-kinase inhibitor (Rho-I) (Y-27632, 1 mM aerosols) prior to the last 8 allergen inhalations and OVA-Rho-I/MCh/constant or DI groups received Y-27632 at the end of allergen sensitization protocol before methacholine challenge. The bronchodilatory effect of DI in guinea pigs that were exposed to methacholine was assessed by using an animal ventilator. The bronchodilatory effect was assessed using several parameters: the airway pressure maintenance, airway pressure recovery, and decline of airway pressure.

Results indicated that application of Y-27632 prior to methacholine challenge reduces the airway smooth muscle ability to maintain pressure and also causes further decline in airway pressure in OVA-sensitized animals undergone DI. However, the inhibition of Rho-kinase before OVA inhalations had minimal effect.

We propose that alteration of Rho-kinase signaling pathway may be one of the mechanisms underlying the impairment of DI-induced bronchodilation in OVA-sensitized guinea pigs.

N-acetylcysteine (NAC) has gained attention recently in dermatology as a unique anti-oxidant. In light of progress in nanotechnological methods, it was hypothesized that loading NAC onto nanofibers would positively affect skin wound healing. The objective of this study was to fabricate NAC-loaded electrospun mats and test their effect on wound healing in vivo and in vitro.      Polyvinyl alcohol (PVA)-based mats loaded with NAC at three concentrations were electrospun and characterized in terms of physicochemical properties and drug release profile. Human fibroblast cells (in vitro) and mouse full-thickness skin wounds (in vivo) were treated with mats for 5 and 14 days, respectively. Wound area, tissue histopathology, fibroblast proliferation and cellular oxidative state were evaluated. Mats containing 5% PVA/NAC showed thinner fibers with suitable physicochemical properties and a sustained drug release profile. PVA/NAC (5%) mats enhanced fibroblast proliferation and attachment in vitro. The mats resulted in significant wound closure with high levels of re-epithelialization and collagen fiber synthesis on day 14 post-surgery in vivo. The mats also reduced granulation tissue and edematous stroma to a higher extent. These findings were accompanied by a significant decrease in tissue lipid peroxidation and higher superoxide dismutase activity, which may explain how NAC improved wound healing. We propose an NAC-loaded nanofibrous mat that takes the advantage of a porous nanoscaffold structure to release NAC in a sustained manner. This mat may be a promising candidate for further clinical evaluation.

This study aimed to examine the effects of genistein and daidzein on endometrial receptivity by histopathological, immunohistochemical, and biochemical techniques. In this study, 72 female Sprague-Dawley rats were randomly divided into 8 groups. The endometrial receptivity model was applied to identified groups. Experimental animals were given periorally 10 mg/kg and high 40 mg/kg doses of genistein and daidzein for 5 days by gavage. At the end of the experiment, uterine tissues were evaluated histopathologically, immunohistochemically, and biochemically. When histopathological findings were examined, significant decreases in pinopod formation were observed in high dose genistein and daidzein groups. When compared with the endometrial receptivity group, immunohistochemical staining findings showed a significant decrease in the expression of integrin β3, integrin αvβ3, LIF, and HOXA10 and an increase in MUC 1 expression in the high dose of genistein and daidzein groups.  In biochemical evaluations, it was determined that genistein and daidzein increased estrogen levels and decreased progesterone levels in a dose-dependent manner.   Genistein and daidzein have a negative effect on endometrial receptivity.  Therefore, individuals with a risk of infertility should pay attention to the consumption of genistein and daidzein.

Apoptosis of pulmonary alveolar septal cells is a pathogenesis characteristic of chronic obstructive pulmonary disease (COPD). Endothelial cell specific molecule-1 (ESM-1) plays an important role in apoptosis of cells. Here, we aimed to determine whether ESM-1 can involve in cell apoptosis in emphysematous mice and stable COPD patients. The sample size of patients was small, so two separate models were studied. At day 0, 11, and 22, murine were injected IP with 0.3 ml of PBS/Cigarette smoke extract, and euthanized at day 28. Lung tissues from 20 stable COPD patients and 12 Controls were evaluated. Serum was obtained from 25 stable COPD patients and 12 healthy Controls. Pulmonary function, pathology, pulmonary apoptosis index (AI), expression of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF) and ESM-1 in lung tissue, and concentration of ESM-1 in serum were tested. Protein expression of ESM-1, VEGF and HGF were decreased significantly in emphysematous mice (p <0.05), while AI was increased (p <0.05). Correlation analysis indicated that association between AI and ESM-1 was negative (p <0.01), VEGF and ESM-1 was positive (p <0.01), and HGF and ESM-1 was positive (p <0.01). In stable COPD patients, we proved that ESM-1, VEGF and HGF were decreased significantly, while AI was increased (p <0.05). Correlation between AI and ESM-1 was negative (p <0.01), VEGF and ESM-1 was positive (p <0.01), and HGF and ESM-1 was positive (p <0.01). ESM-1 expression decreased and AI increased in emphysematous mice and stable COPD patients. Findings suggested that ESM-1 may be involved in anti-apoptotic therapy of COPD.

Various therapeutic approaches, including stem-cell-based strategies and tissue engineering, have been proposed for oral ulcerative lesions. We investigated the effects of adipose tissue-derived stem cells (ADSCs) seeded onto the curcumin-loaded collagen scaffold in the mucosal healing of oral ulcers in rats. The current experimental study was conducted on 40 male Sprague-Dawley rats. Oral ulcers were created over both sides of buccal mucosa, and the rats were randomly divided into four equal groups: 1) an untreated group (negative control); 2) Teriadent-treated group (positive control); 3) group treated with curcumin-loaded collagen scaffold; and 4) group received the ADSCs (3 × 106 cells) seeded onto the curcumin-loaded collagen scaffold. Rats were sacrificed on 3rd and 7th day after ulceration for histopathological examination as well as measurement of tissue levels of myeloperoxidase (MPO), superoxide dismutase (SOD), and Interleukin-1 beta (IL-1β) activity. Compared with the negative control, the tissue levels of MPO and IL-1β were significantly decreased in all treated groups (p <0.0001); however, the SOD activity was elevated (p <0.0001). The highest SOD activity as well as the lowest MPO and IL-1β levels were observed in the ADSCs-curcumin-loaded collagen scaffold group. The ulcer healing process at 3rd and 7th day follow-up was much more progressed in the ADSCs-curcumin-loaded collagen scaffold group in comparison with the untreated group (P=0.037 and P=0.004, respectively). According to the findings of this study, ADSCs seeded onto the curcumin-loaded collagen scaffold seems to have a promising potential for oral ulcer healing applications.

This study aimed to investigate the effect of bee venom, a form of alternative therapy, on rotenone-induced Parkinson’s disease (PD) in mice. Moreover, the possible modulation by bee venom of the effect of L-dopa/carbidopa or rasagiline was examined.

Rotenone (1.5 mg/kg, subcutaneously; SC) was administered every other day for two weeks and at the same time mice received the vehicle (DMSO, SC), bee venom (0.065, 0.13, and 0.26 mg/kg; intradermal; ID), L-dopa/carbidopa (25 mg/kg, intraperitoneal; IP), L-dopa/carbidopa+bee venom (0.13 mg/kg, ID), rasagiline (1 mg/kg, IP) or rasagiline+bee venom (0.13 mg/kg, ID). Then, wire hanging and staircase tests were performed and mice were euthanized and brains’ striata separated. Oxidative stress biomarkers namely, malondialdehyde (MDA), nitric oxide (NO), reduced glutathione (GSH), paraoxonase-1 (PON-1), and total antioxidant capacity (TAC) were measured. Additionally, butyrylcholinesterase (BuChE), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-α), and dopamine (DA) were evaluated. Brain histopathological changes and caspase-3- expression were done.

Bee venom significantly enhanced motor performance and inhibited rotenone-induced oxidative/nitrosative stress, observed as a reduction in both MDA and NO along with increasing GSH, PON-1, and TAC. Besides, bee venom decreased MCP-1, TNF-α, and caspase-3 expression together with an increase in BuChE activity and DA content.

Bee venom alone or in combination with L-dopa/carbidopa or rasagiline alleviated neuronal degeneration compared with L-dopa/carbidopa or rasagiline treatment only. Bee venom via its antioxidant and cytokine reducing potentials might be of value either alone or as adjunctive therapy in the management of PD.

Regarding Lemon verbena gastroprotective effects, we investigated the protective effects of Lemon verbena extracts on reducing gastric ulcer induced by indomethacin.

Rats received aqueous and ethanolic extracts of Lemon verbena (50, 100, and 200 mg/kg), zileuton (100 mg/kg), montelukast (10 mg/kg), or 1% Tween 80 in presence or absence of indomethacin (100 mg/kg).

Indomethacin produced stomach ulcer and increased neutrophils percentage and MDA level compared with the control group (p <0.001). Co-administration of indomethacin and zileuton, montelukast and ethanolic (200 mg/kg) (p <0.001), aqueous extract (200 mg/kg) (p <0.05) reduced ulcer compared with the indomethacin group (p <0.001). Ethanolic extracts (100 and 200 mg/kg) and aqueous extract (200 mg/kg) reduced the MDA level (p <0.001). Ethanolic (50, 100, and 200 mg/kg) and aqueous extracts (200 mg/kg) significantly decreased neutrophils percentage compared with the indomethacin group (p <0.001).

Aqueous and particularly ethanolic extracts of Lemon verbena have protective effects on indomethacin-induced gastric ulcers.