温阳生肌膏及其有效成分改善线粒体功能减轻高糖诱导的血管内皮细胞损伤

Wenyang Shengji Ointment and its active ingredients improve mitochondrial function and alleviate high glucose induced damage to vascular endothelial cells

目的探讨温阳生肌膏及其有效成分(肉桂醛、没食子酸)对高糖状态下血管内皮细胞线粒体损伤的保护作用。方法CCK-8法确定温阳生肌膏有效成分(肉桂醛、没食子酸)干预浓度,将人脐静脉血管内皮细胞(EA-hy926)分为对照组(Con)、高糖组(HG)、温阳生肌膏组(WYSJ)、肉桂醛组(CA)、没食子酸组(GA)。以CCK-8法检测细胞增殖活性,成管实验检测细胞成管能力,透射电镜观察细胞线粒体超微结构,DCFH-DA荧光探针检测细胞活性氧(ROS)含量,比色法检测细胞ATP含量。结果CA组浓度选用10μM,GA组浓度选用5μM。与Con组相比,HG组细胞增殖活性下降(P<0.05);成管能力下降(P<0.05);线粒体肿胀、破裂、数量减少;ROS含量增多(P<0.05);ATP含量减少(P<0.05)。与HG组相比,WYSJ组、CA组、GA组细胞增殖活性加强(P<0.05);成管能力加强(P<0.05);线粒体超微结构损伤改善、数量增加,线粒体自噬增加;ROS含量减少(P<0.05);ATP含量增加(P<0.05)。与CA组和GA组相比,WYSJ组细胞增殖活性较强(P<0.05);成管能力较CA组弱(P<0.05);ROS含量较CA组和GA组多;ATP含量多(P<0.05)。结论温阳生肌膏及其有效成分可能通过清除细胞ROS,改善线粒体功能,增加ATP生成,从而减轻高糖诱导的血管内皮细胞损伤。

Objective To explore the protective effects of Wenyang Shengji Ointment and its active ingredients(Cinnamalde-hyde,Gallic acid)on mitochondrial damage in vascular endothelial cells under high glucose conditions.Methods CCK-8 meth-od was used to determine the intervention concentration of the active ingredients(Cinnamaldehyde,Gallic acid)of Wenyang Shengji Ointment.Human umbilical vein endothelial cells(EA-hy926)were divided into Control group(Con),High glucose group(HG),Wenyang Shengji Ointment group(WYSJ),Cinnamaldehyde group(CA),Gallic acid group(GA).The cell pro-liferation activity was detected by CCK-8 method,the tube forming ability was detected by tube forming experiment,the ultra-structure of mitochondria was observed by transmission electron microscope,the content of reactive oxygen species(ROS)was de-tected by DCFH-DA fluorescent probe,and the content of ATP was detected by colorimetry.Results The concentration of CA Group was 10μM,and that of GA group was 5μM.Compared with the Con group,the cell proliferation activity of HG group de-creased(P<0.05);The tube forming ability decreased(P<0.05);Mitochondria swelled and cristae were broken;ROS con-tent increased(P<0.05);ATP content decreased(P<0.05).Compared with HG group,the cell proliferation activity of WYSJ group,CA Group and GA group was enhanced(P<0.05);The ability of tube formation was strengthened(P<0.05);Mito-chondria swelled,ruptured,and decreased in number,while mitophagy occurred;ROS content decreased(P<0.05);ATP con-tent increased(P<0.05).Compared with CA Group and GA group,the cell proliferation activity of WYSJ group was stronger(P<0.05);The ability of tube formation was weaker than that of CA Group(P<0.05);The content of ROS was higher in the CA group than in the GA group;ATP content was higher(P<0.05).Conclusion Wenyang Shengji Ointment and its active in-gredients may alleviate high glucose induced damage to vascular endothelial cell by scavenging cellular ROS,improving mitochon-drial function,and increasing ATP production.