Megsin基因转染对小鼠肾小球系膜细胞单核细胞趋化蛋白-1及细胞间黏附分子-1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单核细胞趋化蛋白-1(MCP-1)及细胞间黏附分子-1(ICAM-1)表达的影响。方法:高糖环境中培养小鼠肾小球系膜细胞,分别培养12、24、48 h,采用MTT法检测细胞增殖程度,免疫细胞化学和Western blot法检测系膜细胞megsin、MCP-1、ICAM-1蛋白表达水平,ELISA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、MCP-1及ICAM-1表达增强,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,而megsin shRNA质粒转染可明显减弱上述变化。结论:Megsin可上调MCP-1及ICAM-1表达,促进系膜细胞增殖及系膜外基质积聚。

Effects of megsin gene transfection on the expressions of MCP-1 and ICAM-1 in mouse glomerular mesangial cells

AIM: To observe the effects of megsin gene transfection on the expressions of monocyte chemoattractant protein-1(MCP-1) and intercellular adhesion molecule-1(ICAM-1).METHODS: Mouse glomerular mesangial cells were cultured in high glucose medium,and then cell proliferation was measured by MTT assay at 12,24 and 48 h respectively.The expressions of megsin,MCP-1 and ICAM-1 in mesangial cells were detected by immunocytochemical staining and Western blotting.The concentration of type Ⅳ collagen in the culture supernatant of mesangial cells was measured by ELISA.RESULTS: Under high glucose concentration,the expressions of megsin,MCP-1 and ICAM-1 increased;the concentration of type Ⅳ collagen in the cell supernatant was elevated as well;and mesangial cell proliferation was enhanced.After megsin gene transfection,the above changes were more significant,but were abated following megsin shRNA/transfection.CONCLUSION: Megsin gene can up-regulate the expressions of MCP-1 and ICAM-1,promote mesangial cell proliferation and mesangial extracellular matrix accumulation.