高糖对人肾小球系膜细胞增殖及细胞外基质的影响

摘要

Objective To observe the effects of glucose with different concentrations on human mesangial cells ( HRMCs) fibroneet in (Fn) and type collagen IV (Col IV)- Methods In vitro HRMCs were put into different glucose concentrations respectively (5.5, 10, 20, 30 and 40 mmol/L) and processed in order to detect the proliferation of HRMCs with MIT method at different times ( 12, 24, 48, 72 h) and the contents of Fn and Col Ⅳ with ELISA method in 48 hours. Results High glucose could promote cell proliferation after 12, 24, 48, 72 h, and it became stronger as glucose concentration increasing and the time lasting longer. Compared with the control group, 30 mmol/L D-glucose after 24 h, the expression of Fn and Col IV were increased. Conclusions High glucose has a dose and time effect on the prolifer ation of HRMCs, glucose with 30 mmol/L and 48 h is the most appropriate conditions for cellular stimulation. High glucose can promote HRMCs to produce ECM, mesangial cell ECM over expression can directly lead to the accumulation of ECM, and the occurrence of glomeru lar sclerosis.%目的 观察不同浓度的葡萄糖对人肾小球系膜细胞增殖的影响及对细胞外基质纤维连接蛋白(Fn),Ⅳ型胶原(ColⅣ)的影响.方法 体外培养人肾小球系膜细胞株,分别加入葡萄糖终浓度为5.5、10、20、30、40 mmol/L进行处理,于不同时间段(12、24、48、72 h)用MTT法检测肾小球系膜细胞的增殖情况,ELISA法检测培养48 h后条件培养基中Fn及ColⅣ的含量.结果 高糖作用12、24、48、72 h均能促进细胞的增殖,且随着葡萄糖浓度升高作用时间的延长促进作用增强.检测细胞上清液的Fn和ColⅣ.与对照组相比,30 mmol/L D-葡萄糖作用48 h后,Fn和ColⅣ表达增高.结论 高浓度葡萄糖对系膜细胞增殖作用具有量效及时效关系,我们选择30 mmol/L浓度的葡萄糖作用48 h为最合适的细胞刺激条件.高浓度葡萄糖可促进系膜细胞细胞外基质的产生,而系膜细胞细胞外基质过度表达,可以直接导致细胞外基质的积聚,从而导致肾小球硬化的发生.