胡黄连总苷对高糖诱导肾小球系膜细胞氧化应激的保护作用

Protective effect of total glucosides of Picrorhiza scrophulariiflora against oxidative stress in glomerular mesangial cells induced by high glucose

胡黄连总苷(total glucosides of Picrorhiza scrophulariiflora,TGP)是具有抗氧化作用的活性组分。由于氧化应激在糖尿病肾病发病中起关键作用,因此本研究考察了TGP对高糖培养下系膜细胞病变和氧化应激损伤的影响。通过高糖(25 mmol.L-1)刺激3周造成糖尿病系膜细胞损伤模型,分别以荧光素探针DCFH-DA,Rh123和Fluo-3/AM负载细胞,流式细胞仪法检测细胞内活性氧(ROS)、线粒体膜电位(MMP)和[Ca2+]i,观察TGP对高糖引起系膜细胞肥大、细胞外基质分泌增加的保护作用。结果表明,高糖培养组系膜细胞肥大,胶原IV的分泌增加,细胞内ROS升高,MMP降低,[Ca2+]i升高,TGP能明显改善高糖诱导的系膜细胞肥大和降低胶原IV的分泌,降低细胞内ROS含量,提高MMP的水平和降低[Ca2+]i。因此TGP可以保护高糖诱导的肾小球系膜细胞氧化应激损伤。

Total glucosides of Picrorhiza scrophulariiflora （TGP） is the active principal in Picrorhiza scrophulariiflora which has antioxidant effect. Since oxidative stress plays a key role in diabetic nephropathy, we investigated the effects of TGP on oxidative stress in bovine glomerular mesangial cells （MC） induced by prolonged high glucose. Bovine glomerular mesangial cells were cultured and passages 2 -3 were used for the experiment. Mesangial cells were cultured in high glucose medium, and treated with TGP for 3 weeks. Then collagen IV excreted by mesangial cells were detected, and the percentages of cell cycle were observed by flow cytometry technique. The levels of reactive oxygen species （ROS） , mitochondrial membrane potential （MMP） and [ Ca^2＋]i were measured by flow cytometry after loaded with fluorescent probe DCFH-DA, Rh123 and fluo-3-acetoxymethylest. TGP significantly decreased the excretion of collagen IV and cell hypertrophy induced by high glucose, reduced the levels of ROS and [ Ca^2＋]i, and increased MMP. Therefore we conclude that TGP could protect mesangial cells against oxidative stress induced by high glucose.