rho激酶抑制剂法舒地尔对高糖培养ins-细胞影响.docxVIP

华中科技大学硕士学位论文Fasudil Contributes to INS-1 Cells Exposed to Hyperglycemiavia Inhibiting Rho/ Rho-kinase PathwayAbstractObjective:The aim of this study was to investigate the effects of the Rho-kinase inhibitor, fasudil ,on INS-1 cells exposed to hyperglycemia.,and clarify a contribution of the Rho/Rho-kinasepathway to the glucotoxicity in the INS-1 cells.Methods:The mRNA expressions of insulin PDX-1 and Rho-kinase was detected by RT-PCR inthe INS-1 cells of incubation with 5.6mmol/l, 11.1mmol/l and 16.7mmol/l glucose after 48hours. Meanwhile, the mRNA expressions of insulin and PDX-1 were detected by RT-PCRin the INS-1 cells of co-incubation with 16.7 mmol/l glucose and different dose offasudil..The protein expression of PDX-1 was detected by immunohistochemistry in theINS-1 cells of co-incubation with 16.7mmol/l glucose and different dose of fasudil.Results:As compared with 5.6 and 11.1 mmol/ l glucose, the expression of Rho-kinase mRNAsignificantly increased (P0.05) in the INS-1 cells of incubation with 16.7mmol/l glucose. Asexpected, preculture of INS-1 cells in 16.7 mmol/ l glucose determined a marked reduction in theexpression mRNA of both insulin and PDX-1 (P0.05). As compared with 16.7 mmol/lglucose, the presence of fasudil counteracted several deleterious effects of high glucose exposure onINS－1 cells, including reduction of insulin and PDX-1 mRNA expression (P0.05), andincreased PDX-1 protein expression (P0.05).Conclusions:In this reasearch this is the first time to display that: 1. Hyperglycemia up-regulated themRNA expression of Rho-kinase.2. The Rho- kinase inhibitors, and in particular fasudil,protected INS-1 cells from glucotoxicity 3.It is possible for Rho/ Rho-kinase pathway to involvein the glucotoxicity in the INS-1 cells.Key words：glucotoxicity ; insulin；PDX-1； Rho/ Rho-kinase pathway ; fasudil2华中科技大学硕士学位论文前言一.糖尿病与糖毒性随着发病率的迅速增加，糖尿病已成为一个最普遍的威胁人类健康的的代谢性疾病[1,2]。糖尿病主要分为两种类型。1 型糖尿病是一个自身免疫性疾病，细胞介导的自身免疫反应损伤胰腺β细胞,导致胰岛素的绝对不足。2 型糖尿病是一个多基因综合征，其主要特征是胰岛素抵抗和β细胞功能