Andrographolide induced heme oxygenase-1 expression in MSC-like cells isolated from rat bone marrow exposed to environmental stress.

BACKGROUND AND OBJECTIVES: Mesenchymal stem cells (MSC-like cells) are the most important stem cells that are used in transplantation clinically in various applications. The survival rate of MSC-like cells is strongly reduced due to adverse conditions in the microenvironment of transplantation, including environmental stress. Heme oxygenase-1 (HO-1) is a member of the heat shock protein, as well as a stress-induced enzyme, present throughout the body. The present study was conducted to investigate the effect of andrographolide, an active derivative from andrographolide paniculate, on HO-1 expression in mesenchymal stem cells derived from rat bone marrow. MATERIALS AND METHODS: The rat bone marrow-derived mesenchymal stem cells (BMSC-like cells) were extracted and proliferated in several passages. The identity of MSC-like cells was confirmed by morphological observations and differential tests. The flow cytometry method was used to verify the MSC-specific markers. Isolated MSC-like cells were treated with different concentrations of andrographolide and then exposed to environmental stress. Cell viability was assessed using the MTT colorimetric assay. A real-time PCR technique was employed to evaluate the expression level of HO-1 in the treated MSC-like cells. RESULTS: Isolated MSC-like cells demonstrated fibroblast-like morphology. These cells in different culture mediums differentiated into osteocytes and adipocytes and were identified using alizarin red and oil red staining, respectively. As well, MSC-like cells were verified by the detection of CD105 surface antigen and the absence of CD14 and CD45 antigens. The results of the MTT assay showed that the pre-treatment of MSC-like cells with andrographolide concentration independently increased the viability and resistance of these cells to environmental stress caused by hydrogen peroxide and serum deprivation (SD). Real-time PCR findings indicated a significant increase in HO-1 gene expression in the andrographolide-receiving groups (p < 0.01). CONCLUSION: Our results suggest that andrographolide creates a promising strategy for enhancing the quality of cell therapy by increasing the resistance of MSC-like cells to environmental stress and inducing the expression of HO-1.

Combined dandelion extract and all-trans retinoic acid induces cytotoxicity in human breast cancer cells.

Breast cancer is one of the most prevalent and deadly cancers among women worldwide. Recently, natural compounds have been widely used for the treatment of breast cancer. Present study evaluated antiproliferative and anti-metastasis activities of two natural compounds of dandelion and all-trans-retinoic acid (ATRA) in human MCF-7 and MDA-MB231 breast cancer cells. We also evaluated the expression of MMP-2, MMP-9, IL-1ß, p53, NM23 and KAI1 genes. Data showed a clear additive cytotoxic effect in concentrations of 40 µM ATRA with 1.5 and 4 mg/ml of dandelion extract in MCF-7 and MDA-MB231 cells, respectively. In both cell lines, compared with the untreated cells, the expression levels of MMP-9 and IL-1ß were significantly decreased while p53 and KAI1 expression levels were increased. Besides, MMP-2 and NM23 had different expressions in the two studied cell lines. In conclusion, dandelion/ATRA co-treatment, in addition to having strong cytotoxic effects, has putative effects on the expression of anti-metastatic genes in both breast cancer cells.

Effect of hydroalcoholic seed extract of Nigella sativa on hepatic and pancreatic factors of Nrf2 and FGF21 in the regulation of insulin transcription factors of MafA and PDX-1 in streptozotocin-treated diabetic rats.

INTRODUCTION: Nigella sativa (N. sativa), one of the most commonly used medicinal herbs with antioxidant properties, increases blood insulin levels and lowers fasting blood sugar. Nuclear Erythroid Factor-Related Factor 2 (Nrf2) and Fibroblast Growth Factor 21 (FGF21) are two antioxidant factors that are increased by oxidative stress and hyperglycemia. The present study investigated how hydroalcoholic extract of N. sativa seed (HENS) increases blood insulin levels, taking into account changes in antioxidant factors and expression of insulin transcription factors. MATERIALS AND METHODS: Two groups of male diabetic wistar rats were treated orally with HESN at doses of 200 and 400 mg/kg-body weight for one month. Fasting blood sugar (FBS) and insulin were measured using standard kits by photometric and ELISA methods, respectively. The expression levels of the Nrf2, FGF21 and ß-Klotho genes as well as the insulin gene-stimulating transcription factors of MafA and PDX-1 were evaluated using real-time PCR. Oxidative stress was assessed by assessing serum total oxidation status (TOS), malondialdehyde (MDA), and total antioxidant capacity (TAC). RESULTS: HSEN showed a significant reducing effect on FBS and oxidative biomarkers and an increasing effect on serum insulin levels in treated diabetic rats compared to untreated diabetics (P < 0.05). The elevated levels of NRF2 and FGF21 in the liver and pancreas of the diabetic control group were significantly reduced after treatment with both HESN doses (P < 0.05). Following the ameliorative effects of HENS on pancreatic tissue and the reduction of oxidative stress, the expression level of MafA and PDX1 genes approached the level of these factors in healthy rats (P < 0.05). CONCLUSION: This study showed the therapeutic effects of HENS on diabetic pancreas by reducing oxidative stress and tissue damage, modifying the expression levels of PDX-1 and MafA genes, and regulating insulin secretion and blood glucose levels.

Glutamine supplementation can reduce some atherosclerosis markers after exhaustive exercise in young healthy males.

OBJECTIVES: Glutamine can be beneficial to athletes for its antiinflammatory and antioxidant effects. The present study was designed to investigate the effect of glutamine supplementation on some atherosclerosis markers after exhaustive exercise in young healthy males. METHODS: In an intervention study, 30 healthy males (case = 15 and control = 15) were randomly assigned into two groups. For 14 d, the intervention group received 0.3 g of glutamine per kilogram of body weight per day, with 25 g of sugar in 250 mL of water, and the control group received 25 g of sugar per 250 mL of water. At the end of the intervention, the participants completed one session of exhaustive exercise, and then fasting blood samples were taken to test serum levels of atherosclerosis markers. RESULTS: In the intervention group, the serum levels of leptin, cholesterol, and oxidized low-density lipoprotein were lower than in the control group after 2 wk of glutamine supplementation (P < 0.05). Interleukin-6 serum levels were lower in the intervention group compared to the control group after supplementation, but not significantly. Serum levels of leptin, interleukin-6, cholesterol, and oxidized low-density lipoprotein, as well as the ratio of oxidized low-density lipoprotein to high-density lipoprotein, were significantly decreased in the intervention group compared to the control group after exhaustive exercise (P < 0.05). CONCLUSIONS: Glutamine supplementation has beneficial effects for athletes, particularly those doing strenuous physical exercise, through reducing atherosclerosis-related biomarkers and elevating serum adiponectin levels, and it can potentially play a role in decreasing the initiation and progression of atherosclerosis.

Comparison of the effects of zinc oxide and zinc oxide nanoparticles on the expression of hepcidin gene in rat liver.

OBJECTIVES: Nanoparticles have special properties, such as increased intestinal absorption, permeability, and so on. Zinc oxide (ZnO) nanoparticles have medical applications such as using in drug production. Studies of ZnO nanoparticles have shown the role of these particles in reducing or increasing the genes expression. Given the important role of hepcidin in the development of anemia and iron overload diseases, this study investigated the effect of ZnO nanoparticles on the hepatic expression of the hepcidin gene to help find a way to treat these diseases. METHODS: In this experimental study, 24 male Westar rats were divided into three groups: control, ZnO treating group and ZnO nanoparticle treating group. Both ZnO and ZnO nanoparticles were injected with 50 mg/kg body weight for 14 days. At the end, serums were collected and iron, ferritin and IL-6 levels were measured. Expression of the hepcidin gene was done by Real Time PCR. RESULTS: ZnO and the ZnO nanoparticle significantly increased the expression of the hepcidin gene relative to the control group. The increase in expression of the hepcidin gene in ZnO nanoparticles was more significant than in the ZnO. CONCLUSION: ZnO nanoparticles led to significant increase in expression of the hepcidin gene.

The Effect of Glutamine Supplementation on Oxidative Stress and Matrix Metalloproteinase 2 and 9 After Exhaustive Exercise.

BACKGROUND: Glutamine is the most abundant amino acid in plasma and skeletal muscles and an important fuel for immune system cells. It has beneficial anti-inflammatory and antioxidant properties which may be considered as a potentially useful supplement for athletes. The present study was conducted to investigate the effect of glutamine supplementation on oxidative stress and matrix metalloproteinase 2 and 9 after exhaustive exercise in young healthy males. MATERIALS AND METHODS: In this study, 30 healthy males (supplement =15 and control=15) were randomly assigned into two groups. The supplement group received 0.3 g/kg BW of glutamine along with 25 gr of sugar dissolved in 250 cc water per day. The control group received 25 gr of sugar in 250 cc water per day. Fasting blood samples were taken at baseline and at the end of 14 days of intervention. The participants underwent exercise until experiencing full-body exhaustive fatigue for 16 ± 2.84 mins, and then fasting blood samples were taken. Serum levels of TAC, MDA, MMP2, MMP9, glutathione, and hs-CRP were measured. RESULTS: Serum levels of MDA and hs-CRP significantly decreased in the supplement group (p< 0.05). The serum level of TAC significantly increased in the supplement group (p< 0.05). Glutathione serum levels significantly increased after exhaustive exercise (p< 0.05). Serum levels of MMP2 and MMP9 remained unchanged. CONCLUSION: Results of this study showed that, some biochemical factors are time-dependent and can increase or decrease over time, as well as, serum levels of hs-CRP and MDA decreased with glutamine supplementation along with the increase in the TAC serum levels, but this supplementation had no effect on serum levels of MMP2 and MMP9 in exhaustive exercise.

Evaluation of Haptoglobin Genotype and Some Risk Factors of Cancer in Patients with Early Stage Esophageal Cancer.

INTRODUCTION: Esophageal cancer is one of the most lethal gastrointestinal cancers that has a complex and diverse etiology, with several genetic and nutritional factors involved in its etiology. The purpose of this study was to investigate the type of haptoglobin genotype and its relationship with some nutritional and biochemical risk factors affecting the prevalence of esophageal cancer in patients with early stage esophageal cancer. MATERIALS AND METHODS: In this study, 44 patients (20 males and 24 females) with early stage esophageal cancer and 44 healthy subjects, classified as control group, (19 males and 25 females) were selected. Haptoglobin (HP) genotype was determined employing PCR technique. Nutritional data were analyzed using standard food frequency questionnaire (FFQ) method. Serum levels of malondialdehyde (MDA), nitrate and nitrite were measured employing the colorimetric method. Serum levels of p53 protein were measured using the enzyme-linked immunosorbent assay (ELISA) technique. RESULTS: The results of our study showed for the first time that HP1-1 genotype was the most prevalent genotype in esophageal cancer patients in Golestan province, Iran. HP2-2 genotype was the most frequent in the control group. Serum levels of MDA were significantly higher in the patients' group compared to the control group (P˂0.001). Weight and body mass index (BMI) were significantly lower in the patients' group than the control group (P<0.01). Food frequency analysis revealed that the consumption of fruits and vegetables in the patients' group was lower than that of the control group (P<0.05). CONCLUSION: The results of our study showed for the first time that HP1-1 genotype is the dominant genotype in patients with esophageal cancer in Golestan province. As well, modification of nutritional pattern and consumption of high level of antioxidant compounds can be effective in reducing the prevalence of esophageal cancer in this region.

Mesenchymal Stromal Stem Cell-Derived Microvesicles Enhance Tumor Lysate Pulsed Dendritic Cell Stimulated Autologous T lymphocyte Cytotoxicity

Background: Immunotherapy is one promising therapeutic strategy against glioma, an aggressive form of brain cancer. Previous studies have demonstrated that multiple tumor antigens exist and can be used to induce tumor specific T cell responses. Furthermore, recently it was shown that TLR4-primed mesenchymal stem cells (MSCs), also known as MSC1, mostly elaborate pro-inflammatory mediators. Compared to MSCs, MSC-derived microvesicles (MVs) have advantageous properties that present them as stable, long lasting effectors with no risk of immune rejection. Therefore, peripheral blood monocyte derived dendritic cells (MoDCs) have been used to load tumor antigens and stimulate T cell mediated responses in the presence of MSC1-derived MVs in vitro. Methods: The B92 tumor cell line was heated to 43°C for 90 min prior to preparation of tumor cell lysates. MVs were purified by differential ultracentrifugation after isolation, stimulation of proliferation and treatment of MSCs. Autologous T cells isolated from non-adherent cells were harvested during the procedure to generate MoDCs and then incubated with heat stressed tumor cell lysate pulsed DCs in the presence of MSC1-derived MVs. T cells were then co-cultured with tumor cells in 96-well plates at a final volume of 200 µl CM at an effector: target ratio of 100:1 to determine their specific cytotoxic activity. Results: Flow cytometric analysis, T cell mediated cytotoxicity showed that heat stressed tumor antigen pulsed MoDCs and MSC1-derived MVs primed T cells elicited non-significantly enhanced cytotoxic activity toward B92 tumor cells (P≥0.05). Conclusion: These findings may offer new insights into tumor antigen presenting technology involving dendritic cells and MSC1-derived MVs. Further exploration of the potential of such nanoscale particles in immunotherapy and in novel cancer vaccine settings appears warranted.

The effect of conjugated linoleic acid on oxidative stress and matrix metalloproteinases 2 and 9 in patients with COPD.

Background: Natural antioxidants in foods may be used in prevention and treatment of oxidative stress and inflammation in COPD. Therefore, this study aimed to evaluate the effect of conjugated linoleic acid (CLA) supplement as natural antioxidants on oxidative stress levels, and MMP2 and MMP9 serum levels in COPD patients. Materials and methods: This clinical trial study was conducted on 90 (supplement group=45 and control group=45) COPD patients in Ardabil city, Iran, in 2015. After obtaining written consent, general information was collected from each patient using a validated and reliable questionnaire. Supplement group received 3.2 g of CLA and those in the control group were given 3.2 g of placebo for 6 weeks on a daily basis. Fasting blood samples were taken from all of the patients for testing of malondialdehyde (MDA), MMP2, and MMP9 levels at the beginning and end of the study. Data were analyzed using Kolmogorov-Smirnov test, independent samples t-test, paired sample t-test, chi-square test, and ANOVA. Results: There were no significant differences between the two groups with regard to mean age, smoking status, and serum level of MDA at the beginning of the study. In the supplement group, the serum level of MDA decreased significantly at the end of the 6th week compared to that in the beginning of the study (p=0.0004), while in the placebo group, the difference was found to be insignificant. The serum level of MMP9 decreased significantly in the supplement group, while in the placebo group its level increased significantly as compared to that at the beginning of the study (p<0.05). The serum levels of MMP2 indicated no significant differences between the two groups neither at the beginning nor at the end of the study. Conclusion: These findings indicated that CLA supplementation may be helpful for COPD patients through inhibiting the production of oxidative stress and controlling MMP9 serum levels.