megsin基因转染对高糖环境中肾小球系膜细胞增殖和基质金属蛋白酶2及其抑制因子表达的影响

目的 观察megsin基因转染对高糖环境中肾小球系膜细胞基质金属蛋白酶2（MMP-2）和组织金属蛋白酶抑制因子2（TIMP-2）的表达及Ⅳ型胶原水平的影响，探讨megsin与系膜细胞增殖和细胞外基质代谢的关系。 方法 高糖环境中培养小鼠肾小球系膜细胞，分别于培养12、24、48 h末，应用MTT法检测细胞增殖程度；Western印迹法检测系膜细胞megsin、MMP-2、TIMP-2蛋白表达水平；放免法检测细胞培养上清Ⅳ型胶原浓度。 结果 高糖环境中肾小球系膜细胞megsin、TIMP-2表达上调，MMP-2表达下调，细胞增殖明显，细胞上清液中Ⅳ型胶原浓度升高。megsin基因转染后上述变化趋势更加显著。 结论 megsin可诱导系膜细胞增殖，并通过上调TIMP-2、下调MMP-2抑制细胞外基质降解，是加速肾小球硬化的可能机制之一。

Effects of megsin gene transfection on mesangial cell proliferation and expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 under high glucose concentration

Objective To observe the effects of megsin gene transfection on mesangial cell proliferation and expression of matrix metalloproteinase-2(MMP-2),tissue inhibitor of metalloproteinase-2(TIMP-2)and type IV collagen under high glucose concentration,and to investigate the relationship between megsin and mesangial cell proliferation and extracellular matrix metabolism.Methods Mouse glomerular mesangial cells were cultured in high glucose medium,then cell proliferation was measured by MTT at 12,24 and 48 h respectively after culture.Protein levels of megsin,MMP-2,TIMP-2 in mesangial cells were detected by Western blot.Concentration of type Ⅳ collagen in supernatant of mesangial cells was measured by radioimmunochemistry.Results Under high glucose concentration,megsin and TIMP-2 expression were increased,MMP-2 expression was decreased,the concentration of type Ⅳ collagen in the cellular supernatant was increased,and mesangial cell proliferation was enhanced.After megsin gene transfection,the above changes were more significant.Conclusion Megsin gene induces mesangial cell proliferation and inhibits matrix degradation through TIMP-2 up-regulation and MMP-2 down-regulation,which is probably one of the mechanisms of accelerating glomerulosclerosis.