Achillea millefolium L. is cultivated in Iran and widely used in traditional medicine for gastrointestinal disorders. The aim of this study was to determine the effect of hydroalcoholic extract of A. millefolium on the contraction and relaxation of isolated ileum in rat. In this experimental study, aerial parts of A. millefolium were extracted by maceration in ethanol 70% for 72h. Terminal portion of ileum in 100 male Wistar rats was dissected and its contractions were recorded isotonically in an organ bath containing Tyrode solution (37 ºC, pH 7.4) under one gram tension. Acetylcholine (1mM) and KCl (60mM) were used to create isotonic contractions. Propranolol and Nω-Nitro-L-arginine methylester hydrochloride (L-NAME) were used to investigate the mechanisms of action prior to giving the extract to the relevant groups. Data were compared by ANOVA and Turkey's post hoc test.. The results showed that the ileum contraction was induced by KCl and acetylcholine induced contraction was significantly reduced by A. millefolium extract. The cumulative concentrations of A. millefolium relaxed the KCl and acetylcholine induced contractions (n=14, p<0.001). The inhibitory effect of extract on contraction induced by KCl and acetylcholine was not significantly affected neither by propranolol (1μM) nor by L-NAME (100 μM). There was no significant difference in the rate of relaxation by propranolol and L-NAME between the two groups. In conclusion, A. millefolium can inhibit contraction of smooth muscle of ileum in rat, and it can be used for eliminating intestinal spasms. These results suggest that the relaxatory effect of A. millefolium on ileum contractions can be due to the blockade of voltage dependent calcium channels. In addition, the β-adrenoceptors, cholinergic receptors and nitric oxide production are not powerful actors in inhibitory effect of A. millefolium. So, the nitric oxide and adrenergic systems may also be involved in the antispasmodic effect of A. millefolium.
In spite of previous reports, the role of transforming growth factor-β1 (TGF-β1) on cardiomyocyte differentiation, especially in the present autologous serum (AS) in culture medium, is still unclear. So, the purpose of this study was to investigate the potential of rat bone marrow mesenchymal stem cells (rBMSCs) to proliferate and differentiate towards cardiomyocyte lineage with the use of AS. Most expansion protocols use a medium supplemented with fetal bovine serum (FBS) as nutritional supplement. FBS is an adverse additive to cells that are proliferated for therapeutic purposes in humans because the use of FBS carries the risk of transmitting viral and bacterial infections and proteins that may initiate xenogeneic immune responses. Therefore, bone marrow cells were cultured in a medium supplemented with 10% AS, 10% FBS, and serum free medium (SFM). Then, rBMSCs were cultured with TGF-β1 (10 ng/ml) for 2 wk. The number of viable cells in AS and FBS groups were measured with MTT assay. Beating areas frequency, up to fourth week after plating, were monitored and evaluated daily. The characteristics of cardiomyocytes were assessed by semi-quantitative reverse transcription polymerase chain reaction and western blot. MTT result indicated that rBMSCs in AS proliferated markedly faster than FBS and SFM. The number of beating areas significantly increased in AS compared to FBS medium. A noticeable increase in the cardiac genes expression was observed in AS. Moreover, western blot analysis confirmed that cardiac proteins were increased in the AS condition. In conclusion, the present study could be extended toward the safe culture of MSCs for the treatment of heart defects.
PurposeProgesterone has been reported to inhibit the proliferation of breast cancer and osteosarcoma cells; however, its inhibitory mechanism has not yet been clarified. The aim of the present study was to clarify the effects of progesterone on apoptosis in breast cancer (MCF-7) and human osteosarcoma (MG-63) cells.Materials and methodsIn this experimental study the cytotoxic effect of progesterone was measured in MCF-7 and MG-63 cells exposed to different concentrations of progesterone using MTT assay, and effective concentrations were identified. The expression levels of the Bax, P53 and Bcl-2 genes were evaluated by real-time PCR, and caspase-3, 8 and 9 activity levels were determined using a colorimetric method. Hoechst staining and flow cytometry were used to confirm apoptosis. The data were statistically analyzed using one-way analysis of variance (ANOVA) and independent-samples t-test.ResultsCompared to the control group, we observed a significant increase in the expression levels of the Bax and P53 genes and the activity levels of caspase-3 and 9, and a significant decrease in the expression level of the Bcl-2 gene in MCF-7 and MG-63 treated with effective concentration of progesterone. The caspase-8 activity level did not change significantly in treated MG-63 but increased in treated MCF-7 cells. Hoechst staining and flow cytometry results confirmed apoptosis in the cells exposed to effective concentration of progesterone.ConclusionsThe cytotoxic effect of progesterone on breast cancer and osteosarcoma cells was mediated by apoptotic pathways. In this context, progesterone triggers the extrinsic and intrinsic apoptotic pathways in MCF-7 cells and induces the intrinsic apoptotic pathway in MG-63 cells.
Background and Aim :Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders of women in reproductive age. Anovulation and hyperandrogenism due to PCOS are major causes of infertility. The aim of the present study was to evaluate the combined effect of Nigella Sativa hydro-alcoholic extract and honey on gonadotropins and sex hormones level inpolycystic ovary syndrome in rats. Materials and Methods: 40 adult Wistar rats, weighting 200 -220 g were divided randomly into 5 groups: including intact, Sham (letrozole solvent), PCOS and 2 experimental PCOS groups. Rats were treated with letrozole for 21 days to induce PCOS. In the experimental groups, PCOS rats were treated with 2 doses of combination 300mg/kg Nigella sativa extract with 1200mg/kg honey and 600mg/kg Nigella sativa extract with 2400mg/kg honey for 28 days. The serum levels of LH, FSH, estrogen, progesterone and testosterone were measured by ELISA method and each serum were analyzed using ANOVA and TUKEY statistical tests (P<0.05). Results: A significant decrease was seen in serum levels of LH, estrogen and testosterone (P<0.05) in PCOS group treated with maximum dose of combination of N. sativa extract and Honey compared with the PCOS group. Whereas, a significant increase in serum levels FSH in PCOS group treated with maximum dose and progesterone in the PCOS group treated with maximum and minimum dose of combination of N. sativa extract and Honey compared with the PCOS group (p< 0.05). Conclusion: Beneficial effects of combination of N. sativa extract and Honey was seen but further study is needed.
Objectives: In diabetes mellitus type 1, beta cells are mostly destroyed; while in diabetes mellitus type 2, beta cells are reduced by 40% to 60%. We hope that soon, stem cells can be used in diabetes therapy via pancreatic beta cell replacement. Induced pluripotent stem cells are a kind of stem cell taken from an adult somatic cell by "stimulating" certain genes. These induced pluripotent stem cells may be a promising source of cell therapy. This study sought to produce isletlike clusters of insulin-producing cells taken from induced pluripotent stem cells. Materials and Methods: A human-induced pluripotent stem cell line was induced into isletlike clusters via a 4-step protocol, by adding insulin, transferrin, and selenium (ITS), N2, B27, fibroblast growth factor, and nicotinamide. During differentiation, expression of pancreatic β-cell genes was evaluated by reverse transcriptase-polymerase chain reaction; the morphologic changes of induced pluripotent stem cells toward isletlike clusters were observed by a light microscope. Dithizone staining was used to stain these isletlike clusters. Insulin produced by these clusters was evaluated by radio immunosorbent assay, and the secretion capacity was analyzed with a glucose challenge test. Results: Differentiation was evaluated by analyzing the morphology, dithizone staining, real-time quantitative polymerase chain reaction, and immunocytochemistry. Gene expression of insulin, glucagon, PDX1, NGN3, PAX4, PAX6, NKX6.1, KIR6.2, and GLUT2 were documented by analyzing real-time quantitative polymerase chain reaction. Dithizonestained cellular clusters were observed after 23 days. The isletlike clusters significantly produced insulin. The isletlike clusters could increase insulin secretion after a glucose challenge test. Conclusions:This work provides a model for studying the differentiation of human-induced pluripotent stem cells to insulin-producing cells.
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