【摘要】： 糖尿病是由多種病因引起的以慢性高血糖為特征的代謝性疾病，嚴(yán)重危害著人類健康。目前糖尿病的治療手段有胰島素注射，胰腺和胰島移植，細(xì)胞治療等。由于長(zhǎng)期注射胰島素存在自體耐受和較高的并發(fā)癥，胰島移植面臨著供體不足和長(zhǎng)期使用免疫抑制劑的問(wèn)題。因此，干細(xì)胞以其多向分化潛能成為糖尿病治療的一條很好的途徑。 本實(shí)驗(yàn)以人脂肪來(lái)源基質(zhì)細(xì)胞（human adipose-derived stromal cells,hADSCs）為研究對(duì)象，利用堿性成纖維細(xì)胞生長(zhǎng)因子、胰高血糖素樣肽-1（GLP-1）、肝腦勻漿因子和尼克酰胺等在體外將其誘導(dǎo)分化為胰島素分泌細(xì)胞，通過(guò)形態(tài)學(xué)、RT-PCR、雙硫腙染色、免疫細(xì)胞化學(xué)等方法，，對(duì)誘導(dǎo)后的細(xì)胞進(jìn)行分泌功能和活性測(cè)定，探討脂肪來(lái)源基質(zhì)細(xì)胞分化為胰島素分泌細(xì)胞的潛能及其分化機(jī)制。結(jié)果顯示，體外hADSCs在適當(dāng)葡萄糖濃度，勻漿因子、GLP-1和尼克酰胺共同作用下，可以分化為胰島素分泌細(xì)胞，雙硫腙染色呈陽(yáng)性，RT-PCR檢測(cè)到胰島素基因的表達(dá)，而且外環(huán)境中葡萄糖濃度變化影響著胰島素的分泌。利用誘導(dǎo)后的細(xì)胞分泌胰島素的特性，為今后干細(xì)胞臨床治療糖尿病提供一定的理論和實(shí)踐依據(jù)。
[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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R329

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