Iranian Journal of Basic Medical Sciences
Volume:17 Issue: 4, Apr 2014

The goal of this study was assessing the prophylactic effect of exercise and its role as an adjuvant therapy on level of cytokines involved in angiogenesis in estrogen-dependent breast cancer. Forty female BALB/c mice were randomly assigned to exercise-tumor-exercise (ETE), exercise-tumor-rest (ETR), rest-tumor-exercise (RTE) and rest-tumor-rest (RTR) groups. After orientation in the environment, two groups of mice performed continuous endurance exercise for 8 weeks, and thereafter estrogen-dependent MC4L2 cancer cells were injected to them. Then, one group of each of trained and non-trained mice performed endurance exercise 5 days per week for 6 weeks. Tumor volume was measured by a digital caliper weekly. Finally, the mice were sacrificed; tumor tissue was removed, immediately frozen and kept in -70°C. Tumor sample was homogenized; levels of cytokines were measured and quantified using ELISA. There was significant reduction in the level of interlukin-6 (IL-6) (P=0.001), Vascular endothelial growth factor (VEGF) (P=0.0001) and tumor volume (P=0.0001) among the groups performing endurance exercise after malignancy (RTE and ETE) in comparison with groups not performing endurance exercise (ETR and RTR), and these results were in agreement with tumor growth rate. Exercise can cause reduction in levels of pro-inflammatory cytokines in tumor tissue. Decreased IL-6 production could reduce the generation of VEGF, resulting in reduced intra-tumor angiogenesis. Due to reduction of the level of these cytokines in groups doing exercise before and after malignancy, exercise is presumed to be an adjuvant therapy in estrogen-receptor dependent tumors in addition to its effective prophylactic role.

Consumption of high-fat foods is one of the major causes of obesity. Physical exercise is a strategy used to counteract obesity. The aim of this study was to investigate the effect of eight weeks endurance training and high-fat diet (HFD) on appetite-regulating hormones in rat plasma. Twenty eight male Wistar rats were randomly divided into four groups: Control group with standard diet (CSD), endurance training with a standard diet (ESD), control group with high-fat diet (CHFD) and endurance training with high-fat diet (EHFD). Twenty-four hr after the last training session, the blood samples were obtained and analyzed for hormones levels. The significant increased weight gain and food intake and decreased plasma nesfatin-1 and PYY3-36 levels were observed in CHFD group, while exercise under the HFD antagonized these effects. There were no significant changes in ghrelin, insulin and leptin levels in different groups. These results suggest that exercise can prevent fattening effect of HFD. Probably, performing exercise makes a reduction of food intake and weight gain in rat via the increase in nesfatin-1 and PYY levels. However, further studies are necessary to understand the exact mechanisms involved in this field.

Bradykinin is a part of the kinin-kallikreinsystem which is involved in ischemia-reperfusion injury via B1 and B2 receptors.Noscapine is a non-competitive antagonist of bradykinin receptors. Noscapine has been reported to to be able to protect some organs against ischemia-reperfusion injury but its effect on renal ischemia-reperfusion injury (RIR) in rats is unknown. Therefore, the present study was designed to evaluate the effect of noscapine on renal ischemia-reperfusion injury in rats. Twenty four rats were randomly assigned to four groups; sham, RIR control, pre-and post-treatment with noscapine. To induce RIR injury, 20 days after right nephrectomy, animals underwent a midline laparotomy and the renal artery was clamped for 40 min to induce ischemia, and the clamp was then removed to allow reperfusion for 48 hr. Animals received noscapine or vehicle 1 hr before RIR or just prior to reperfusion. At the end of the experiment, animals were killed by cardiac exsanguination. Blood samples were collected to assess blood urea nitrogen (BUN) and creatinine. The kidneys were also removed for histopathlogical and western-blot analysis. Noscapine treatment 1 hr before RIR or just prior to reperfusion protects the renal tissue structure as compared with the control. The expression levels of the studied inflammatory mediators, TNF-α and MCP-1in pretreated-, and treated-noscapine groups decreased as compared with the control group. The levels of BUN and creatinine in pre-, and post-treated noscapine groups were significantly lower than in control animals. Noscapine protects renal tissue structure and function against RIR through down-regulation of the inflammatory mediators.

Regulatory T cells, including CD4+CD25+Fox3+ and CD8+CD28- cells play an important role in regulating the balance between immunity and tolerance. Since multiple sclerosis is an inflammatory autoimmune disease, regulatory T cells are considered to be involved in its pathogenesis. In this study, we investigated the circulatory numbers of the two mentioned types of regulatory T cells and also their association with different clinical characteristics in 84 multiple sclerosis patients. 84 patients with multiple sclerosis and 75 normal individuals were studied. Demographic and clinical information of all participants were collected via questionnaire and clinical examination as well as MRI. The peripheral blood frequency of two different subgroups of regulatory T cells (CD4+ CD25+Foxp3+ and CD8+CD28- cells) were analyzed by flow cytometry using anti-human antibodies conjugated with CD4-FITC / CD25-PE/Foxp3-PE-Cy5, CD3-PE/CD8a-PE-Cy5/CD28-FITC. The frequency of CD4+CD25+Foxp3+ cells in multiple sclerosis patients was significantly less than that in healthy controls (P=0.006) and in mild forms less than that in sever forms (P=0.003). There was not any correlation between the frequency of regulatory T cells and different clinical variables. Our results showed that the number of CD4+CD25+Foxp3+ cells decreases significantly in multiple sclerosis patients, which probably shows the regulatory role of these cells in multiple sclerosis.

Cefixime (Cfx), is a semi-synthetic third-generation oral cephalosporin antibiotic that is prescribed for the treatment of susceptible infections. There are some procedures for the determination of Cfx in pharmaceutical formulations and biological samples. Herein a spectrofluorimetric method was proposed for Cfx determination based on the fluorescence quenching of terbium-danofloxacin (Tb3+-Dano) in the presence of Cfx. Cfx was detected based on fluorescence quenching of terbium-danofloxacin (Tb3+-Dano) in the presence of Cfx with maximum excitation and emission wavelengths at 347 nm and 545 nm, respectively. The quenched fluorescence intensity of Tb3+- Dano system is proportional to the concentration of Cfx. The optimum conditions for the determination of Cfx were studied. The maximum response was achieved under optimum conditions of [Tris buffer]= 0.008 mol/l (pH 6.5), [Tb3+]=1×10-4 mol/l and [Dano]=1×10-4 mol/l. The developed method was evaluated in terms of accuracy, precision and limit of detection. The linear concentration ranges for quantification of Cfx were 8.8×10-8-8.8×10-7 mol/l and 1.1×10-7-8.8×10-7 mol/l in standard and human serum samples with the detection limits (S/N=3) of 2.8×10-8 mol/l and 3.9×10-8 mol/l, respectively. The Cfx was determined in pharmaceutical tablets and spiked serum samples and the results were satisfactory. This method is simple, practical and relatively interference-free for determination of Cfx in pharmaceutical tablets and serum samples.

In insulin resistance, the insulin action in liver, muscles and adipocytes decreases and result in hyperglycemia, dyslipidemia and hyperinsulinemia. In this study we evaluate the effect of Zataria multiflora extract on insulin sensitivity in high fructose fed insulin resistant rats, since this extract was shown antihyperglycemic effect in streptozotocin induced diabetes in rats. Experimental rats were fed with high fructose diet for 6 weeks and then were treated with Z. multiflora extractor a pioglitazone solution for 2 weeks. Blood and tissue samples were collected for analysis at the end of two weeks. Blood glucose, serum level of triglyceride and cholesterol were measured by auto analyzer. Insulin and adiponectin levels were assayed by enzyme-linked immunosorbent assay (ELISA) method. Plasma free fatty acids profile was studied by gas chromatography. Peroxisome proliferator activated receptor (PPAR.γ) and Glucose transporter type 4 (GLUT.4) gene expressions were assessed by real time polymerase chain reaction (PCR) and western blotting. Animals were treated by Z. multiflora extractshowed insulin (43±11pmol/l), adiponectin (5.3±0.5 μg/ml), glucose (144±9.8 mg/dl), and triglyceride (120±10 mg/dl) levels significantly improved as compare with the control group [insulin (137±34 pmol/l), adiponectin (3.9±0.15 μg/ml), glucose (187±15mg/dl), and triglycerides (217±18 mg/dl)]. PPARγ protein level, also significantly increased in Zataria multiflora treated group. This study demonstrates the beneficial effects of Zataria multiflora extract on insulin resistance in rats fed with a high-fructose diet through at least three mechanisms including direct insulin like effect, increasing in adiponectin and of PPARγ protein expression.

Morphine is widely used to treat chronic pain. However, its utility is hindered by the development of tolerance to its analgesic effects. Despite the renowned beneficial effects of physical exercise on cognitive functions and signs of morphine withdrawal in morphine-dependent rats, little is known about the roles of voluntary and forced exercises in tolerance to analgesic effect of morphine in rats. In this study, rats were injected with 10 mg/kg of morphine, once daily, SC over a period of 8 days of either voluntary or treadmill exercise. Following these injections, the percent of maximum possible effect (%MPE) of morphine was measured on the 1st, 4th, and 8th days by hot plate test. Both voluntary and forced exercises significantly increased pain threshold compared to the sedentary group (P<0.05). Voluntary and forced exercises also significantly increased potency of morphine compared to sedentary morphine group (P<0.05). Thus, we concluded that voluntary and forced exercises blocked the development of tolerance during 8 daily simultaneously treatments. When exercising rats were returned to sedentary conditions, sensitivity to the analgesic effects of morphine increased significantly and persisted during sedentary period in the exercising rats. In other words, %MPE of the exercising morphine-group increased significantly compared to saline group (P<0.05). Our results showed that voluntary and forced exercises may be possible methods for treating the development of tolerance to analgesic effect of morphine in rats.

In the past few decades, variety of foetal, embryonic and adult stem and progenitor cells have been tried with conflicting outcome for cell therapy of central nervous system injury and diseases. Cellular characteristics and functional plasticity of Globose basal stem cells (GBCs) residing in the olfactory epithelium of rat olfactory mucosa have not been studied in the past by the neuroscientists due to unavailability of specific markers for GBCs. In the present research, we standardized some techniques to isolate GBCs from rat olfactory epithelium in pure form using a highly selective GBC-III antibody passaged through fluorescence activated cell sorter (FACS). We also characterized these cells immunohistologically using various pluripotent stem cell markers. This work also throws some light on ionic channels present on these stem cells which are responsible for their neuron induction potential. Globose basal stem cells were isolated from rat olfactory epithelium using GBC-III antibody and were characterized as multipotent stem cells using various neural progenitor markers. Ionic channels on GBCs were studied with voltage clamping. GBCs could be isolated in pure (99% purity) form and were found to be stained positive for all neural progenitor cell markers. Voltage gated Na+ channels were completely absent, which proves the unexcitable nature of GBCs. Leaky K+ channels were found to be present on the GBC which was of no significance. This research work can be helpful in understanding the nature [T1] of these stem cells and utilising them in future as potent candidates for neuro-regenerative therapies.

Gentamicin sulphate (GS) nephrotoxicity seems to be related to the generation of reactive oxygen species. There is evidence that oxygen preconditioning increases the activity of antioxidant enzymes. Forty eight female rats were divided into 6 groups (n=8) as follows: group 1 was the control, group 2 received daily GS, groups 3,4 and 5 received oxygen 2 hr/day for 2 days, 4 hr/day for 2 days, 4 hr/day for 4 days, recpectively and then received daily GS, group 6 received oxygen 2 hr/day for 2 days and then received 2 hr oxygen before daily GS injection. Oxygen (with 90% purity) used at the flow rate of 4 l/min. GS administred for 8 days (100 mg/kg, IP). Tissue sections prepared from the left kidney, stained with PAS method and then studied hisopathologically and stereologically. The right kidneys were homogenized and the supernatants were prepared. Serum MDA, creatinine and urea, renal MDA, gluthatione and catalase activity were measured. The data were analyzed by Mann-Whitney U test at the significant level of P Oxygen therapy significantly improves serum creatinine and urea, preserve tubular volume density, reduce tubular necrosis in groups 4 and 6 compared to group 2. Oxygen therapy significantly increases renal catalase in groups 4 and 6 compared to group 2. Pretreatment with normobaric hyperoxia and daily oxygen therapy improved gentamicin nephrotoxicity possibly via inhibition of lipid peroxidation and increasing the renal catalase activity but could not restore any parameter at the same levels as control group.

This study investigated the effects of melatonin, famotidine, mirtazapine, and thiamine pyrophosphate on ischemia/reperfusion (I/R) injury in diabetic rats and evaluated oxidant and antioxidant marker measurement results. It also examined the effects of the drugs aimed at preventing infertility that may result from I/R injury. Diabetic rats were divided into a control group (IRC) to be exposed to I/R, an ovarian I/R + 2.2 mg/kg melatonin (IRML) group, an ovarian I/R + famotidine (IRFA) group, an ovarian I/R + 20 mg/kg mirtazapine (IRMR) group, an ovarian I/R + 20 mg/kg thiamine pyrophosphate (IRTP) group, and a sham operation (SO) group. In the control group exposed to I/R, the levels of the oxidant parameters Malondialdehyde (MDA) and Myeloperoxidase(MPO) were significantly higher compared with the SO group, while the levels of the antioxidant parameters glutathione (GSH), Glutathioneperoxidase(GPO), Glutathione reductase (GSHRd), Glutathione S - transferase (GST), and[y1] Superoxide dismutase (SOD) were significantly lower. Melatonin, famotidine, mirtazapine, and thiamin pyrophosphate prevented a rise in oxidant parameters and a decrease in antioxidants in ovarian tissue exposed to I/R. However, apart from thiamin pyrophosphate, none of the drugs were able to prevent infertility caused by I/R injury. Prevention of ovarian I/R injury-related infertility in rats with induced diabetes is not through antioxidant activity. Thiamine pyrophosphate prevents infertility through an as yet unknown mechanism. This study suggests that thiamine pyrophosphate may be useful in the prevention of I/R-related infertility in diabetics.

Cervical cancer is a malignancy that is the second most common cause of death from cancer in women throughout the world. Paederus beetle (Paederus fuscipes) extract (PBE), contains bioactive compounds such as pederine which has cytotoxic properties and blocks DNA and protein synthesis at very low concentrations. In this investigation we tried to determine the effects co-treatment with PBE and gamma irradiation on HeLa cells. The viability of the cells was measured by two MTT and Colony assay. We found that supplementing gamma irradiation therapy with PBE does not increase cell death and it might even interfere with its cytotoxicty at the concentrations below 0.1 ng/ml and the viability for irradiation vs irradiation + PBE was 37%: 60%. This finding might be due to radioprotective effects of the very low doses of PBE against gamma radiation.

Bacopa monnieri (BM),a traditional ayurvedic medicine,is a well-known memory enhancer. We have explored the role of BM against decabrominated diphenyl ether (PBDE-209)-induced alterations in neonate and young female mice. Mice were orally administered with B. monnieri at the doses of 40, 80 or 120 mg/kg body weight along with PBDE-209 (20 mg/kg body weight) from postnatal day (PND) 3-10. Levels of malondialdehyde, protein carbonyl and activities of superoxide dismutase and glutathione peroxidase were measured at both ages. The correct choices and reference/working memory errors of young mice were evaluated by Morris water and radial arm maze. The results showed that BM at the dose of 120 mg/kg significantly (P<0.05) restored the levels of oxidants and the activities of antioxidant enzymes in frontal cortex and hippocampus of neonates against PBDE-209-induced toxicity. BM plays a neuroprotective role against PBDE-209-induced alterations in oxidative status.