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近年来许多研究表明糖尿病肾病肾小球硬化与高血糖状态有关。有研究显示人类肾小球硬化区主要为正常细胞外基质如层粘连蛋白Laminin(LAM)和Ⅳ型胶原CollagenⅣ(C-Ⅳ)的过度积聚[Kidney Int,1992,41:556],但糖尿病肾病肾小球硬化机理尚未明了。我们通过同质大鼠肾小球系膜细胞培养,观察高浓度葡萄糖对细胞增殖及细胞外基质合成的影响,探讨糖尿病肾病发病机理。一、材料和方法 1.按经典方法培养。鉴定SD大鼠肾小球系膜细胞,选取第3～5代细胞用于本实验。 2.将上述细胞以1×10~4个·ml~(-1)/孔种于24孔板中,于0.5%FCS RPMI 1640细胞培养液中培养,同时加入不同浓度葡萄糖,取不传代培养后24小时、48
In recent years, many studies have shown that diabetic nephropathy glomerulosclerosis and hyperglycemia state. Studies have shown that human glomerular sclerosis is mainly caused by excessive accumulation of normal extracellular matrix such as Laminin (LAM) and Collagen IV (C-IV) [Kidney Int, 1992, 41: 556], but diabetic nephropathy The mechanism of glomerulosclerosis is still unknown. We through the homogeneous rat mesangial cells cultured to observe the effect of high glucose on cell proliferation and extracellular matrix synthesis, explore the pathogenesis of diabetic nephropathy. First, materials and methods 1. According to the classic method of training. Identification of glomerular mesangial cells in SD rats, select the 3rd to 5th generation cells for this experiment. 2. The above cells in 1 × 10 ~ 4 · ml -1 / well in 24-well plates in 0.5% FCS RPMI 1640 cell culture medium, while adding different concentrations of glucose, take non-subculture After 24 hours, 48