丹酚酸B对高糖诱导的肾小球系膜细胞表型转化及细胞外基质分泌的影响

摘要

目的:研究丹酚酸对高糖诱导的肾小球系膜细胞表型转化及细胞外基质分泌的影响及其机制.方法:培养人肾小球系膜细胞 (HGMCs) , 随机分为正常对照组、高糖组及高糖+丹酚酸B高、中、低剂量组, 高糖组和丹酚酸B各组用含高浓度 (33.3 mmol/L) 葡萄糖的培养基培养72 h, 丹酚酸B各组同时加人相应浓度丹酚酸B共同孵育.Western blot法检测α-平滑肌肌动蛋白 (α-SMA) 的表达水平, ELISA法检测细胞Ⅰ型胶原 (ColⅠ) 、Ⅲ型胶原 (ColⅢ) 、纤维连接蛋白 (FN) 和层粘连蛋白 (LN) 的分泌水平, Western blot法检测转化生长因子β1 (TGF-β1) 的表达及Smad2和p38丝裂原活化蛋白激酶 (MAPK) 的磷酸化水平.结果:高糖孵育72 h后, 肾小球系膜细胞α-SMA的蛋白表达水平明显升高, ColⅠ、ColⅢ、FN及LN蛋白的分泌水平显著增加 (P <0.01) , TGF-β1的表达及Smad2、p38 MAPK的磷酸化水平也明显升高 (P <0.01) ;与丹酚酸B共同孵育可明显降低α-SMA蛋白的表达水平, ColⅠ、ColⅢ、FN和LN的分泌明显减少, TGF-β1的表达及Smad2、p38 MAPK的磷酸化水平显著下降 (P <0.01或P <0.05) .结论:丹酚酸B可明显抑制高糖诱导的肾小球系膜细胞表型转化, 减少ColⅠ和ColⅢ等细胞外基质分泌, 其机制与抑制TGF-β1/Smad信号通路及p38 MAPK活化有关.%AIM:To investigate the effect of salvianolic acid B (Sal B) on high glucose-induced phenotypic transition and extracellular matrix (ECM) secretion in human glomerular mesangial cells (HGMCs) and the underlying mechanisms.METHODS:HGMCs were randomly divided into control group, high glucose group and high glucose plus high dose, medium dose and low dose of Sal B groups.The HGMCs except those in control group were exposed to high glucose (33.3 mmol/L) for 72 h, while those in Sal B groups were co-incubated with indicated concentrations of Sal B.The protein levels ofα-smooth muscle actin (α-SMA) , transforming growth () and phosphorylated Smad2 and p38 mitogen-activated protein kinase (MAPK) were determined by Western blot.The secretion levels of collagen type I (Col I) , collagen type III (Col III) , fibronectin (FN) and laminin (LN) were measured by ELISA.RESULTS:Exposure to high glucose markedly increased the protein expression ofI, Col III, FN and LN in the HG-MCs (P<0.01).The phosphorylation levels of Smad2 and p38 MAPK were also significantly increased (P<0.01).Coincubation with Sal B evidently decreased the protein expression ofI, Col III, FN and LN in the HGMCs induced by high glucose (P<0.05 or P<0.01).The phosphorylated levels of Smad2 and p38 MAPK were also reduced noticeably (P<0.05 or P<0.01).CONCLUSION:Sal B significantly suppresses high glucose-induced phenotypic transition and ECM secretion in the HGMCs, which might be attributed, at least partly, to inhibition ofSmad signaling pathway and p38 MAPK activation.