| Background:Diabetes are one kind of severely impair human health disease, and its prevalence rate rises unceasingly. At present, the insulin substitution treatment was still the main method to treat diabetes. How can permanently cure diabetes becomes one of puzzled questions in endocrine domain. Diabetes mellitus type 1 and 2 (T1DM and T2DM) are complex multifactorial diseases. Loss ofβcell function caused by reduced secretory capacity and enhanced apoptosis is a key event in the pathogenesis of both diabetes types. The transplantiotion of islet cells can treat 1 and 2 diabetes, but the lack of donor and excessiveβcell apoptosis make it hard to be put into use in routine clinical practices. Bone marrow derived cells have the ability of self-renewal and multi-potential of differentiation. In recent years, some researchs demonstrated that the transplantion of bone marrow mesenchymal stem cells (BMSCs) might ameliorate the condition of diabetes animal model, which offered a novel way to cure diabetes. Some researchs have showed that MSCs trans-differentiation into insulin producing cells (IPCs) may partly account for the therapeutic action. However, also some research indicated that little evidence was proved trans-differentiation of bone marrow-derived cells into pancreatic beta cells. Moreover, it cannot explain the disease amelioration after transplantation, regardless in time limit or extent. Some research had confirmed that MSCs may secrete a series of cytokines and the growth factors (GF), these factors play the protective function to periphery cells, which is called paracrine mechanisms. Recent studies have showed that paracrine effects play a significant part in the recreate and and repair of tissues. Therefore, we surmise that paracrine is the mechanism of MSCs'effect to diabetic animals.Objective:To investigate the effect of BMSCs on hydrogen peroxide (H2O2)—induced apoptosis in INS—1 cell by establish aβcell apoptosis model, and the possible mechanism involved.Methods:Isolation and culture BMSCs under sterile conditions from healthy male rats about 180～220 g. INS—1 cells induced by H2O2 to establish theβcell apoptosis model. The apoptotic INS—1 cells were co-cultured with BMSCs. The experiment consisted of 4 groups:Group A: INS—1 cells culture group (Control group); Group B:H2O2 treatment group; Group C:INS—1 cells and BMSCs coculture group; Group D:INS—1 cells and BMSCs coculture with H2O2 treatment group; Group E:iNOS inhibitor 1400W 40μmol/L and H2O2 treatment group; Group F:NOS inhibitor 1400W 80μmol/L and H2O2 treatment group. Cell viability was mesured by MTT. Annexin V—PI doubled staining of flow cytometry was used to quantify apoptosis ratio. NO production was determined by Griess method.Research Results:INS—1 cell viability was reduced by H2O2 in a dose-dependent manner. The cell viability was obviously decreased when the concentration of H2O2 in 40μmol/L or more. With Transwell co-culture, H2O2 induced INS—1 cell apoptosis ratio was significantly lower than in H2O2 group (P<0.05),also the production of NO was decline sharply compared with the group B (P=0.01), which effection was similar with that of 1400W.Conclusion:BMSCs can protect INS—1 cells from apoptosis, induced by H2O2. The mechanism my may be related to the paracrine mechanisms of BMSCs through inhibiting iNOS-NO pathway. The simulation results are valuable theoretical basis for further research on prevention and control diabetes by the utilization of BMSC. |
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