芪参益气滴丸对过氧化氢所致体外心肌细胞损伤的保护作用

PROTECTION OF QISHENYIQIWAN ON CARDIOMYOCYTE INJURY INDUCED BY HYDROGEN PEROXIDE IN VITRO

目的:探讨芪参益气滴丸对过氧化氢所致心肌细胞损伤的保护作用及其机制。方法:采用纯化培养的心肌细胞建立体外过氧化氢损伤模型。实验分5组,每组8孔细胞(24孔细胞培养板):正常对照组;过氧化氢损伤模型组;芪参益气滴丸高、中、低3个不同剂量组。其中损伤模型组过氧化氢浓度为200μmol·L-1。芪参益气滴丸组在200μmol·L-1过氧化氢损伤的基础上,分别加入15、30、45μg·L-1芪参益气滴丸。加入过氧化氢损伤的同时加入芪参益气滴丸,共同孵育12h后测定结果。利用试剂盒分别测定细胞培养液上清中乳酸脱氢酶(LDH)、肌酸磷酸激酶(CPK)和心肌细胞内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)及还原型谷胱甘肽(GSH)的含量。结果:损伤模型组细胞培养液中LDH,CPK含量明显高于正常对照组(P<0.05)和芪参益气滴丸组(P<0.05);损伤模型组心肌细胞内SOD,CAT,GPx和GSH的含量,均明显低于正常对照组(P<0.05)和芪参益气滴丸组(P<0.05)。结论:芪参益气滴丸可明显减轻体外培养条件下过氧化氢所致心肌细胞损伤。

Objective： To study the protection of Qishenyiqiwan on cardiomyocyte injury induced by hydrogen peroxide in vitro. Method： Hydrogen dioxide injured model was implemented with purified cultured cardiomyocytes, and the experiment was divided into 5 groups with 8 pores in each group （24-well cell culture plate） ： normal control group, hydrogen peroxide （200 μmol L- 1 ）injured group, three different Qishenyiqiwan dose（ 15, 30, 45 μg L -1 ） groups, For Qishenyiqiwan groups, the cell culture was damaged by hydrogen peroxide while adding Qishenyiqiwan, and the incubation time was 12 h. The cell culture medium was used for assaying the activity of lactate dehydrogenase （LDH） , creatine phosphokinase （ CPK）, antioxidant enzymes superoxide dismutase （ SOD）, Catalase （ CAT）, glutathione-peroxidase （GPx）, and glutathione （GSH）. Cultured myocardial cells apoptosis was detected by flow cytometry. Results： Compared with the hydrogen peroxide injured group, thee activities of marker enzymes, LDH, CPK in the cell culture medium of the normal control group and Qishengyiqiwan group were significantly decreased（P 〈 0.05） , while the activities of SOD, CAT, GPx and GSH were significantly increased（ P 〈0.05 ） , and apoptosis ratio was decreased markedly（ P 〈0.05 ）. Conclusion： The present study reveals that Qishenyiqiwan can significantly reduce myocardial damaged by Hydrogen dioxide in vitro.