Iranian Journal of Basic Medical Sciences
Year:2015 | Volume:18 | Issue:10|Papers Count:15

Objective(s): this study focused on the effect of waterpipe smoke exposure toxicity on the structure of albino rat’s ventricular tissue and their recovery.Materials and Methods: Albino rats were divided into three groups: control, flavored, and unflavored.The control group was exposed to normal air while the flavored and unflavored groups were exposed to waterpipe smoke for a period of 90 days. Each group was followed by a period of 90 days of fresh air exposure. Following each period, the ventricular tissue was removed for biochemical and histopathological studies.Results: The ventricular tissues of waterpipe exposed rats showed some degree of separation between cardiac muscle fibers, infiltration of lymphocytes, and congestion of blood vessel. Also, thin cross sections of ventricular cells revealed pleomorphic mitochondria with partially disrupted cristae, partial disruption of the myofibrils, and deposited toxic materials. The unflavored waterpipe has more deleterious effects on heart ventricular tissues than the flavored one. Waterpipe smoke didn't induce apoptosis in the ventricular tissue. We also found very high levels of plasma thiocyanate after exposure to smoke in the flavored and unflavored groups, while the control group showed no increase. After the recovery period, those tissues showed partial recovery.Conclusion: Waterpipe smoke induces structural changes in the heart ventricle tissues, causing a negative impact on the capacity of the cardiac muscle for pumping blood and may lead to heart attack due to accumulation of free radicals and tissue inflammation. Cessation of smoking is important in returning most of these changes to their normal structure.

Objective(s): The aim of the present study was to evaluate antioxidant and methyl donor effects of betaine in cerebellum following levodopa and benserazide administration in rats.Materials and Methods: Sprague-Dawley male rats were treated with levodopa (LD), betaine (Bet), levodopa plus betaine (LD/Bet), levodopa plus benserazide (LD/Ben), levodopa plus betainebenserazide (LD/Bet-Ben), and the controls with vehicle for 10 consecutive days, orally.Results: Treatment of rats with LD and benserazide significantly increased total homocysteine in plasma of the LD/Ben group when compared to the other groups. Lipid peroxidation of cerebellum increased significantly in LD‐treated rats when compared to the other groups. In contrast, glutathione peroxidase activity and glutathione content in cerebellum were significantly higher in the betainetreated rats when compared to the LD and LD/Ben groups. Serum dopamine concentration increased significantly in LD‐treated rats in comparison with the LD/Ben group. LD/Bet-treated rats also demonstrated significantly higher dopamine levels when compared to the LD/Ben group.Conclusion: We observed valuable effects of Bet in combination with LD and benserazide, which routinely were used for Parkinson’s disease (PD) treatment, in experimentally-induced oxidative stress and hyperhomocysteinemia in rats. Therefore, it seems that Bet is a vital and promising agent regarding PD for future clinical trials in humans.

Objective(s): In this study the effect of crocin, a carotenoid isolated from saffron, on malathion (an organophosphate insecticide) induced depressive-like behavior in subacute exposure was investigated. Moreover the molecular mechanism of malathion induced depressive-like behavior and its decreasing effect on the level of brain derived neurotrophic factor (BDNF) in rat hippocampus and cerebral cortex were evaluated.Materials and Methods: Male Wistar rats were exposed to malathion (50 mg/kg/day, IP) alone or in combination with crocin (10, 20 and 40 mg/kg/day, IP), imipramine (20 mg/kg/day, IP) and vitamin E (200 mg/kg, three times a week, IP) respectively for 14 days. The forced swimming test (FST) was performed on days 1st, 7th and 14st. The level of malondealdehyde (MDA) and reduced glutathione (GSH) were measured in cerebral cortex and hippocampus of rats. The protein level of BDNF was evaluated using Western blot analysis.Results: Malathion (50 mg/kg, IP) increased immobility time in the FST, without affecting total locomotor activity in open‐field test. Malathion increased the malondealdehyde (MDA) and decreased the glutathione (GSH), whereas these effects were reversed by crocin and vitamin E.Malathion decreased plasma acetylcholinesterase activity, however this effect was not reversed by crocin or vitamin E. Malathion reduced the protein level of BDNF in rat hippocampus. Imipramine and crocin prevented the decreasing effect of malathion on BDNF.Conclusion: These results showed that crocin attenuates some neurochemical and behavioral effects induced by malathion. This neuroprotective effect of crocin may be in part due to its effect on BDNF.

Objective(s): Nitric oxide (NO), a product of inducible nitric oxide synthase (iNOS), contributes in germ cell apoptosis. This study was aimed to evaluate the effects of Aloe vera gel (AVG) on male Wistar rat reproductive organ, serum NO level, and expression of iNOS gene in leydig cells.Materials and Methods: Adult male Wistar rats (n=36) were used for experiments in three groups. The experimental groups were orally administered with the AVG extract solution once-daily as follow: 150 mg.kg-1; group A, 300 mg.kg-1; group B, and only normal saline; group C (control group). They were mated with untreated females and the reproductive and chemical parameters were assessed for each group, including semen quality, serum testosterone, sperm fertility, gonad and body weight, serum NO concentration (by the Griess method), and iNOS gene expression (using RT-PCR).Results: The testes weight, serum testosterone, as well as sperm count and fertility of the AVG treated groups were significantly reduced when compared to the control (P<0.001). Concentration of serum NO was significantly increased (37.1±4.63 mM) in the administrated group with higher AVG concentration, compared to the control group (P<0.001; 10.19±0.87 mM); however, iNOS mRNA expression was increased in the treated animals (P<0.001).Conclusion: iNOS may play a functional role in spermatogenesis via apoptosis, reducing sperm count, but further studies are needed to illustrate the mechanisms by which AVG exerts its negative effects on spermatogenesis and sperm quality.

Objective(s): Angiotensin II type 1 receptor blockers (ARBs) represent one of the widely used antihypertensive agents. In addition to anti‐hypertension effect, some ARBs also show other molecular effects such as activating peroxisome proliferator-activated receptor-g and so on. Here we studied the effects of telmisartan on the rat isolated mesenteric artery rings pre-contracted by phenylephrine (PE).Materials and Methods: Rat mesenteric artery rings were pre-contracted with 10 mM PE, and cumulative concentration-response curves to telmisartan were obtained. The endotheliumdependent mechanisms were investigated by mechanical removal of the endothelium. K+channels were investigated by pretreatment of the artery rings with various K+channel blockers.Results: Telmisartan produced concentration-dependent relaxation of the artery rings precontracted by 10 mM PE. Denudation of the endothelium did not affect the relaxant effect of telmisartan. Pretreatment with BaCl2 nearly inhibited the relaxation induced by the 0.5, 1, 5 and 10 mM telmisartan, but did not affect the relaxation induced by the 50 and 100 mM telmisartan. While the relaxation induced by telmisartan was not affected by pretreatment with TEA, 4-AP and glibenclamide.Conclusion: These findings demonstrated that telmisartan produces concentration dependent vasodilation in isolated rat mesenteric artery rings with or without endothelium pre‐contracted by PE. KIR channel may be involved in such a relaxant effect of telmisartan.

Objective(s): Dispersive liquid-liquid microextraction coupled with gas chromatography (GC)-flame ionization detector was developed for the determination of valproic acid (VPA) in human plasma.Materials and Methods: Using a syringe, a mixture of suitable extraction solvent (40 ml chloroform) and disperser (1 ml acetone) was quickly added to 10 ml of diluted plasma sample containing VPA (pH, 1.0; concentration of NaCl, 4% (w/v)), resulting in a cloudy solution. After centrifugation (6000 rpm for 6 min), an aliquot (1 ml) of the sedimented organic phase was removed using a 1-ml GC microsyringe and injected into the GC system for analysis. One variable at a time optimization method was used to study various parameters affecting the extraction efficiency of target analyte. Then, the developed method was fully validated for its accuracy, precision, recovery, stability, and robustness.Results: Under the optimum extraction conditions, good linearity range was obtained for the calibration graph, with correlation coefficient higher than 0.998. Limit of detection and lower limit of quantitation were 3.2 and 6 mg/ml, respectively. The relative standard deviations of intra and inter-day analysis of examined compound were less than 11.5%. The relative recoveries were found in the range of 97 to 107.5%. Finally, the validated method was successfully applied to the analysis of VPA in patient sample.Conclusion: The presented method has acceptable levels of precision, accuracy and relative recovery and could be used for therapeutic drug monitoring of VPA in human plasma.

Objective(s): Nucleus Raphe Magnus (NRM) that is involved in the regulation of body temperature contains nitric oxide (NO) synthase. Considering the effect of NO on skin blood flow control, in this study, we assessed its thermoregulatory role within the raphe magnus.Materials and Methods: To this end, tail blood flow of male Wistar rats was measured by laser doppler following the induction of hypothermia.Results: Intra-NRM injection of SNP (exogenous NO donor, 0.1- 0.2 ml, 0.2 nM) increased the blood flow. Similarly, unilateral microinjection of glutamate (0.1- 0.2 ml, 2.3 nM) into the nucleus increased the blood flow. This effect of L-glutamate was reduced by prior intra NRM administration of NO synthase inhibitor NG-methyl L-arginine or NG‐nitro-L-arginine methyl ester (L-NAME, 0.1 ml, 100 nM).Conclusion: It is concluded that NO modulates the thermoregulatory response of NRM to hypothermia and may interact with excitatory amino acids in central skin blood flow regulation.

Objective(s): This study focused on determining cytotoxic effects of copper oxide nanoparticles (CuO NPs) on chronic myeloid leukemia (CML) K562 cell line in a cell‐specific manner and its possible mechanism of cell death. We investigated the cytotoxicity of CuO NPs against K562 cell line (cancerous cell) and peripheral blood mononuclear cell (normal cell).Materials and Methods: The toxicity was evaluated using cell viability, oxidative stress and apoptosis detection. In addition, the expression levels of P53, Caspase 3, Bcl-2, and Bax genes in K562 cells were studied by reverse transcription polymerase chain reaction (RT-PCR) analysis.Results: CuO NPs exerted distinct effects on cell viability via selective killing of cancer cells in a dosedependent manner while not impacting normal cells in MTT assay. The dose-dependent cytotoxicity of CuO NPs against K562 cells was shown through reactive oxygen species (ROS) generation. The CuO NPs induced apoptosis was confirmed through acridine orange and propidium iodide double staining.Tumor suppressor gene P53 was up regulated due to CuO NPs exposure, and increase in Bax/Bcl-2 ratio suggested mitochondria‐mediated pathway is involved in CuO NPs induced apoptosis. We also observed that Caspase 3 gene expression remained unchanged up to 24 hr exposure.Conclusion: These molecular alterations provide an insight into CuO NPs-caused inhibition of growth, generation of ROS, and apoptotic death of K562 cells.

Objective(s): Serratia marcescens is one of the nosocomial pathogen. The ability to form biofilm is an important feature in the pathogenesis of S. marcescens. The aim of this study was to determine the anti-adhesive properties of a biosurfactant isolated from Lactobacillus acidophilus ATCC 4356, on S. marcescens strains.Materials and Methods: Lactobacillus acidophilus ATCC 4356, was selected as a probiotic strain to produce biosurfactant. Anti-adhesive activities was determined by pre-coating and co-incubating methods in 96-well culture plates.Results: The FTIR analysis of derived biosurfactant revealed the composition as protein component.Because of the release of such biosurfactants, L. acidophilus was able to interfere with the adhesion and biofilm formation of the S. marcescens strains. In coincubation method this biosurfactant in 2.5 mg/ml concentration showed anti‐adhesive activity against all tested strains of S. marcescens (P<0.05).Conclusion: Our results show anti-adhesive properties of L. acidophilus biosurfactant will be useful against microorganisms responsible for infections in the urinary, vaginal and gastrointestinal tracts, as well as skin, making it a suitable alternative to conventional antibiotics.

Objective(s): EMAP-like Protein 5 (EML5) is a new echinoderm microtubule‐associated protein that is expressed in the central nervous system. The aim of this study was to investigate the expression profile of EML5 in the anterior temporal neocortex of patients presenting with intractable epilepsy (IE).Materials and Methods: Western blot assays were performed to determine EML5 expression in 36 surgically resected anterior temporal neocortices of patients with IE and eight control tissues.Immunohistochemistry and immunofluorescence were employed to explore protein expression in IE.Results: EML5 was highly expressed in both neurons and glial cells of the anterior temporal neocortex of IE patients, whereas only low levels of EML5 were detected in control brain tissues. Western blotting showed an enhanced expression of EML5 protein in the anterior temporal neocortex of IE (optical density (OD)=1.8030±0.1335/1.1852±0.2253, P<0.05) compared with normal control tissues.Conclusion: The results demonstrate that highly expressed EML5 in the neurons and glial cells of the cortex of patients with epilepsy is associated with microtubular dysfunction after frequent and recurrent seizures.

Objective(s): Auraptene (7-geranyloxycoumarin) (AUR), from Citrus species has shown antiinflammatory, neuroprotective, and acetylcholinesterase (AChE) and beta-secretase inhibitory effects.Scopolamine is a nonselective muscarinic receptor antagonist which causes short‐term memory impairments and is used for inducing animal model of Alzheimer’s disease (AD). This research aimed to investigate the effect of AUR on scopolamine-induced avoidance memory retention deficits in stepthrough task in mice.Materials and Methods: The effect of four‐day pre-training injections of AUR (50, 75, and 100 mg/kg, subcutaneous (SC)) and scopolamine (1 mg/kg, IP), and their co-administration on avoidance memory retention in step‐through passive avoidance task, was investigated by measuring the latency to enter to the dark chamber.Results: Pre-training administration of AUR caused significant increase in step‐through latency in comparison with control group, 48, 96, and 168 hr after training trial. The findings of this study showed that scopolamine (1 mg/kg, IP, for four consecutive days) impaired passive avoidance memory retention compared to saline-treated animals. Step-through passive avoidance task results showed that AUR markedly reversed scopolamine-induced avoidance memory retention impairments, 24 and 168 hr after training trial in step-through task.Conclusion: Results from co-administration of AUR and scopolamine showed that AUR reversed scopolamine-induced passive avoidance memory retention impairments.

Objective(s): MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate gene expression.They have important roles in kidney development, homeostasis and disease, and participate in the onset and progression of tubulointerstitial sclerosis and end‐stage glomerular lesions that occur in various forms of chronic kidney disease (CKD). In the present study, we elucidated the role of microRNA 205 (miR‐205) in cisplatin-induced renal cell apoptosis and explored the molecular mechanisms.Materials and Methods: The chronic interstitial nephropathy rat model was induced, and the miRNA expression profile in the kidney cells from rats with CKD was screened. Cisplatin-induced apoptosis in normal renal HK-2 cells was evaluated using flow cytometry, and regulation of miR‐205 on target gene was validated using luciferase assay, western blot and real time PCR assays.Results: We found that miR-205 expression was significantly decreased in the cells from kidney of CKD rat (P<0.01). Our data showed that when miR-205 was overexpressed or silenced using the mimic or inhibitor, the percentages of apoptotic cells were suppressed or increased significantly (P<0.05), respectively. Moreover, we have identified CMTM4 gene, which is involved in cell proliferation and apoptosis, as a novel target for miR-205. In addition, miR-205 could inhibit apoptosis by binding to the 3’UTR of CMTM4 mRNA and inhibiting its transcriptional activity.Conclusion: This study elucidated that miR‐205 plays an important role in the regulation of apoptosis in renal cells, suggesting a potential therapeutic target to hinder CKD development.

Objective(s): MiRNAs have been proposed to be key regulators of tumorigenesis, progression and metastasis. However, their effect and prognostic value in gastric cancer is still poorly known.Materials and Methods: Gastric cancer cell lines were cultured. Tissue samples obtained from 36 gastric cancer patients were used for quantitative real‐time PCR (qRT-PCR) analysis. The tissue microarrays (TMAs) consisted of 126 cases of gastric carcinoma that were used for In situ hybridisation (ISH). Lentivirus plasmids were co‐transfected into 293FT cells. Cell migration was examined using wound‐healing assays. Statistical analyses were performed using SPSS16.0 software.Results: In this study, we found that the expression levels of miR-493 were strongly down‐regulated in gastric cancer and were associated with clinical stage and the presence of lymph node metastases.Moreover, miR-493 might independently predict OS and RFS in gastric cancer. We further found that up-regulation of miR-493 inhibited the proliferation and metastasis of gastric cancer cells, in vitro and in vivo. In addition, miR‐493 directly targeted RhoC, which resulted in a marked reduction of the expression of mRNA and protein. This effect, in turn, led to a decreased ability of growth, invasion and metastasis in gastric cancer cells.Conclusion: Taken together, our findings demonstrate that miR-493 is important for gastric cancer initiation and progression and holds promise as a prognostic biomarker to predict survival and relapse in gastric cancer. It is also a potential therapeutic tool to improve clinical outcomes in this disease.

Objective(s): Lead (Pb) is a toxic metal inducing many destructive effects leading to a broad range of physiological, biochemical, and neurological dysfunctions in humans and laboratory animals.Materials and Methods: Here, we investigated the effect of chronic exposure to Pb (50 mg/l) on oxidative stress, hepatotoxicity, nephrotoxicity, and lipid profile of two different age groups of female rats treated with Pb from delivery until puberty period (40 days, Pb40) and post puberty period (65 days, Pb65).Results: Our results clearly show that the administration of Pb produces oxidative damage in liver and kidney, as strongly suggested by the significant increase in TBARS, decrease in total SH, and the alteration of SOD activity. Elevation in liver function biomarkers, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and reduction in total protein (liver and plasma) and albumin are evidence of perturbations of liver synthetic function. In young Pb-treated group, Pb-induced nephropathy was more pronounced by the increase in the levels of creatinine, urea, and uric acid.However, hyperlipidemia was evident for both Pb-exposed groups leading to a potential risk for cardiovascular diseases and atherosclerosis.Conclusion: It is concluded that Pb induces metabolic and oxidative disturbances depending on the age of the animals, which are not negligible.

Objective(s): Salvia miltiorrhiza (SM) is a popular and classic herb in traditional Chineses medicines. The objective is to confirm the effects of aqueous extract of S. miltiorrhiza (ESM) and its main ingredient on the promotion of orthodontic tooth movement and healing of periodontal ligament in rat.Materials and Methods: Male Sprague-Dawley rats (n=150) were divided into five groups: model control group (0.5 ml/kg phosphate-buffered saline (PBS) injection), ESM group (0.75 g/kg/day of crude drugs) and Danshensu subgroups (250, 500, 750 mg/kg/day of body weight). All rats were administered intramuscularly into the buccal vestibular mucosa of first molar of left maxillary. The indicators such as the moving distance of orthodontic tooth, nuclear factor κB ligand (RANKL) and osteoprotegerin (OPG) expression and osteoclasts were tested.Results: The expressions of RANKL and OPG in the treatment groups were obviously enhanced compared with control group (P<0.05). The increase rate of OPG expression was slower than that of RANKL. But, RANKL decreased conspicuously after no orthodontic pressure was applied, especially in the treatment groups (Danshengsu high dose group at day 30: 2.17 versus 3.47 of control, P<0.01). ESM groups promoted osteoclasts proliferation in the first 20 days.Conclusion: There is a relationship between RANKL/OPG ratio and the number of osteoclasts. ESM might accelerate periodontal alteration of rat orthodontic tooth via producing more osteoclasts.