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目的:观察EGR-1基因转染对高糖环境中小鼠肾小球系膜细胞TGF-β及PDGF-B表达的影响,探讨EGR-1在糖尿病肾病发病机制中的作用。方法:高糖环境中培养小鼠肾小球系膜细胞,采用LipofectamineTM2000瞬时转染EGR-1质粒,于培养12、24、48小时末,应用MTT法检测细胞增殖程度,免疫细胞化学和Western印迹法检测系膜细胞EGR-1、TGF-β及PDGF-B蛋白表达水平,酶联免疫吸附法(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EGR-1、TGF-β及PDGF-B表达增强,细胞增殖明显,细胞上清液中Ⅳ型胶原浓度升高,EGR-1基因转染后上述变化较高糖组更加显著。结论:EGR-1可上调TGF-β及PDGF-B表达,促进系膜细胞增殖及系膜外基质积聚,是加速肾小球硬化的可能机制之一。
Objective: To observe the effect of EGR-1 gene transfection on the expression of TGF-β and PDGF-B in mouse glomerular mesangial cells in high glucose environment and to explore the role of EGR-1 in the pathogenesis of diabetic nephropathy. METHODS: Mouse glomerular mesangial cells were cultured in a high glucose environment and transiently transfected with EGR-1 plasmid by LipofectamineTM2000. At the end of 12, 24 and 48 hours, the cell proliferation, immunocytochemistry and Western blotting were detected by MTT assay The expressions of EGR-1, TGF-β and PDGF-B in mesangial cells were detected by enzyme-linked immunosorbent assay (ELISA). Results: The expression of EGR-1, TGF-β and PDGF-B in glomerular mesangial cells was increased under high glucose condition with obvious cell proliferation and the concentration of type Ⅳ collagen in supernatant of cells increased. Changes in the higher sugar group more significant. CONCLUSION: EGR-1 can up-regulate the expression of TGF-β and PDGF-B and promote the proliferation of mesangial cells and extracellular matrix accumulation. It is one of the possible mechanisms of accelerating glomerulosclerosis.