胡黄连总苷对高糖培养系膜细胞内非酶糖基化和氧化应激的影响

 目的 研究胡黄连总苷（ TGP ）对高糖培养下系膜细胞内非酶糖基化终末产物（ AGEs ）形成和氧化应激的影响，并探讨 AGEs 形成与氧化应激的关系。 方法 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，用荧光素探针 DCFH-DA 和 Rh123 分别负载细胞，流式细胞仪法检测细胞内活性氧（ ROS ）和线粒体膜电位（ MMP ）， HPLC 流动注射法检测系膜细胞内 AGEs 含量，以氨基胍 (AG) 和抗氧化剂二甲亚砜（ DMSO ）为对照，研究 TGP 对氧化应激和 AGEs 形成的作用，及对细胞周期和细胞外基质增加的影响。 结果 高糖培养组系膜细胞停滞于 G1 期，分泌胶原 Ⅳ 增加，细胞内 ROS 和 AGEs 含量增加， MMP 降低， AG 、 DMSO 和 TGP 均能抑制系膜细胞内 ROS 的产生和 AGEs 的形成，增加 MMP ；并改善高糖诱导的系膜细胞肥大和降低胶原 Ⅳ 的分泌。 结论 在糖尿病并症的发病过程中非酶糖化和氧化应激关系密切，称为“糖化氧化”， TGP 通过抑制高糖诱导的糖化氧化从而保护系膜胞损伤。

Effect of Total Glucoside of Picrorhiza scrophulariflora on Nonenzymatic Glycosylation in Glomerular Mesangial Cells Induced by High Glucose

OBJECTIVE To investigate the effects of TGP on advanced glycation end products( AGEs) and oxidative stress in bovine glomerular mesangial cells cultured in high glucose media and discuss the relationship between intracellular AGEs and oxidative stress. METHODS Bovine glomerular mesangial cells were cultured. Mesangial cells were cultured in high glucose media for 3 weeks to make mesangial cells injury models, and treated with TGP in the meantime. AG and DMSO were treated as control drugs. Then collagen Ⅳ excreted by mesangial cells were detected, and the percentage of cell cycle were observed by flow cytometry technique. The levels of reactive oxygen species(ROS) and mitochondrial membrance potential (MMP) were evaluated by flow cytometric analysis after fluorescent probe DCFH-DA and Rhodamine 123 were loaded. Cellular AGEs were measured by HPLC using a flow injection system. RESULTS TGP significantly decreased the excretion of collagen Ⅳ and cell hypertrophy induced by high glucose, reduced the levels of ROS, inhibited the formation of AGEs and increased MMP in mesangial cells. CONCLUSION The formation of AGEs and oxidative stress accelerate each other in the pathogenesis of diabetic complications, which is called “glycoxidation”. TGP can protect mesangial cells by inhibiting glycoxidation induced by high glucose.