人脂肪基质细胞向胰岛素分泌细胞分化诱导的研究

发布时间：2018-10-08 18:03

【摘要】： 糖尿病是由多种病因引起的以慢性高血糖为特征的代谢性疾病，严重危害着人类健康。目前糖尿病的治疗手段有胰岛素注射，胰腺和胰岛移植，细胞治疗等。由于长期注射胰岛素存在自体耐受和较高的并发症，胰岛移植面临着供体不足和长期使用免疫抑制剂的问题。因此，干细胞以其多向分化潜能成为糖尿病治疗的一条很好的途径。本实验以人脂肪来源基质细胞（human adipose-derived stromal cells,hADSCs）为研究对象，利用碱性成纤维细胞生长因子、胰高血糖素样肽-1（GLP-1）、肝脑匀浆因子和尼克酰胺等在体外将其诱导分化为胰岛素分泌细胞，通过形态学、RT-PCR、双硫腙染色、免疫细胞化学等方法，，对诱导后的细胞进行分泌功能和活性测定，探讨脂肪来源基质细胞分化为胰岛素分泌细胞的潜能及其分化机制。结果显示，体外hADSCs在适当葡萄糖浓度，匀浆因子、GLP-1和尼克酰胺共同作用下，可以分化为胰岛素分泌细胞，双硫腙染色呈阳性，RT-PCR检测到胰岛素基因的表达，而且外环境中葡萄糖浓度变化影响着胰岛素的分泌。利用诱导后的细胞分泌胰岛素的特性，为今后干细胞临床治疗糖尿病提供一定的理论和实践依据。
[Abstract]:Diabetes mellitus (DM) is a metabolic disease characterized by chronic hyperglycemia, which is a serious hazard to human health. The current treatment of diabetes is insulin injection, pancreas and islet transplantation, cell therapy and so on. Due to autologous tolerance and high complications of long-term insulin injection, islet transplantation is faced with donor shortage and long-term use of immunosuppressants. Therefore, stem cells with their multi-differentiation potential become a good way to treat diabetes. In this study, human adipose derived stromal cells (human adipose-derived stromal cells,hADSCs) were induced to differentiate into insulin-secreting cells by basic fibroblast growth factor, glucagon like peptide 1 (GLP-1), liver and brain homogenate factor and nicotinamide in vitro. The secretory function and activity of adipose derived stromal cells were determined by means of morphology RT-PCR, dithizone staining and immunocytochemistry. The potential of adipose derived stromal cells to differentiate into insulin-secreting cells and the mechanism of differentiation were discussed. The results showed that hADSCs could differentiate into insulin-secreting cells under proper glucose concentration, homogenate factor GLP-1 and nicotinamide, and the expression of insulin gene was detected by RT-PCR with positive staining of dithizone. And the change of glucose concentration in external environment affects insulin secretion. The characteristics of insulin secretion by induced cells provide a theoretical and practical basis for the future clinical treatment of diabetes by stem cells.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

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