摘要
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 effects of EGR-1 gene transfection on the expression of TGF-β and PDGF-B in mouse glomerular mesangial cells under high glucose, and to investigate the role of EGR-1 in the pathogenesis of diabetic nephropathy. Meth- ods：Mouse glomerular mesangial cells were cultured in high glucose medium and EGR-1 plasmids were transient transfected with lipo- feetamine 2000 reagent, then cell proliferation was measured by MTT at 12, 24 and 48 h respectively. The expression of EGR-1, TGF- β and PDGF-B in mesangial cells were detected by immunocytochemical staining and Western blot, the concentration of type IV collagen in supernatant of mesangial cells was measured by ELISA. Results： Under high glucose concentration, the expression of EGR-1, TGF-β and PDGF-B were increased, the concentration of type IVcollagen in the cellular supernatant was increased, and mesangial cell pro- liferation was enhanced. After EGR-1 gene transfeetion, the above changes were more significant than the high glucose group. Conclu- sion：EGR-1 gene can up-regulate the expression of TGF-~ and PDGF-B, induce mesangial cell proliferation and mesangial extracellular matrix accumulation, which is probably one of the mechanisms of accelerating glomerulosclerosis.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2012年第6期497-501,共5页
Chinese Journal of Immunology
