论文部分内容阅读
目的研究辛伐他汀对体外培养的人腹膜间皮细胞(HPMCs)在高糖刺激下主要细胞外基质Ⅰ型胶原,及其降解酶系基质金属蛋白酶-1(MMP-1)和基质金属蛋白酶抑制剂-1(TIMP-1)分泌与表达的影响。方法胰蛋白酶消化法从人腹膜组织中分离间皮细胞进行原代培养并传代;待细胞同步后,经2.5%高糖和不同浓度的辛伐他汀干预48 h。半定量RT-PCR检测细胞内Ⅰ型胶原、MMP-1和TIMP-1 mRNA表达情况;ELISA方法检测细胞上清液中Ⅰ型胶原、MMP-1和TIMP-1的蛋白质水平。结果辛伐他汀显著下调高糖致HPMCsⅠ型胶原和TIMP-1表达,且在蛋白质和基因水平均呈剂量依赖关系(P<0.01);同时,高剂量的辛伐他汀能显著增加高糖环境下MMP-1蛋白质含量(P<0.01),并上调MMP-1 mRNA表达(P<0.01)。结论辛伐他汀能减少HPMCs高糖刺激下Ⅰ型胶原的合成,并通过调节MMP-1/TIMP-1的平衡,促进Ⅰ型胶原降解,从而减少细胞外基质的沉积。
Objective To investigate the effects of simvastatin on the expression of type I collagen, the major extracellular matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase (MMP) in human peritoneal mesothelial cells (HPMCs) 1 (TIMP-1) secretion and expression. Methods The mesothelial cells were separated from human peritoneal tissue by trypsin digestion method and subcultured. After the cells were synchronized, they were intervened by 2.5% high glucose and different concentrations of simvastatin for 48 hours. Semi-quantitative RT-PCR was used to detect the mRNA expression of type I collagen, MMP-1 and TIMP-1. The protein levels of type I collagen, MMP-1 and TIMP-1 were detected by ELISA. Results Simvastatin significantly down-regulated the expression of type I collagen and TIMP-1 in HPMCs induced by high glucose, and showed a dose-dependent relationship between protein and gene level (P <0.01). Meanwhile, high dose of simvastatin could significantly increase the expression of type I collagen and TIMP- MMP-1 protein (P <0.01), and up-regulated the expression of MMP-1 mRNA (P <0.01). Conclusion Simvastatin can reduce the synthesis of type I collagen in HPMCs stimulated by high glucose and promote the degradation of type I collagen by regulating the balance of MMP-1 / TIMP-1, thereby reducing the deposition of extracellular matrix.