【摘要】 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(25mmol·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。更多还原

【Abstract】 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.更多还原