加味丹参饮抑制p38MAPK表达保护缺氧/复氧乳鼠心肌细胞损伤的实验研究

陈　聪，廖　菁，李　鑫，宋厚盼，黄政德*（湖南中医药大学，湖南长沙410208）



加味丹参饮抑制p38MAPK表达保护缺氧/复氧乳鼠心肌细胞损伤的实验研究

陈聪，廖菁，李鑫，宋厚盼，黄政德*
（湖南中医药大学，湖南长沙410208）

〔摘要〕目的观察加味丹参饮含药血清对缺氧/复氧（hypoxia/reoxygenation，H/R）乳鼠心肌细胞p38 MAPK蛋白表达的影响，以探讨其保护心肌的作用机制。方法将20只SD乳鼠的心肌细胞进行原代培养，建立H/R模型并随机分为H/R组、SB203580（p38MAPK阻断剂组）、加味丹参饮含药血清组，正常心肌细胞组作为对照组。采用T淋巴细胞化学染色法鉴定心肌细胞，罗丹明B（Rhodamine B）染色法观察细胞形态，MTT比色法检测含药血清对乳鼠心肌细胞的毒性，Western-blot法检测MKK3（促分裂原活化蛋白激酶-激酶3）、MKK6（促分裂原活化蛋白激酶-激酶6）、p38MAPK、phospho-p38MAPK蛋白表达。结果与正常血清对照组比较，H/R组MKK3、MKK6的蛋白表达增多，p38MAPK通路阻断剂SB203580和加味丹参饮均不能减少其表达；p38MAPK、phospho-p38MAPK在H/R时表达均增加（P＜0.01），而SB203580和加味丹参饮含药血清均能减少其表达（P＜0.01）。结论加味丹参饮含药血清预处理可通过抑制缺氧，复氧心肌细胞p38MAPK信号通路发挥保护作用，抑制p38MAPK表达及其磷酸化，减轻心肌细胞损伤，起到保护心肌细胞的作用。

〔关键词〕加味丹参饮；心肌细胞；缺氧/复氧；p38MAPK信号通路；促分裂原活化蛋白激酶-激酶3；促分裂原活化蛋白激酶-激酶6；丹参；檀香；赤芍

Experimental Study of Modified Jiawei Danshen Decoction on the Protection of Hypoxia/Reoxygenation Myocardial Cells Injury by Inhibition of p38MARK Expression in Neonatal Rats

CHEN Cong, LIAO Jing, LI Xin, SONG Houpan, HUANG Zhengde*
（Hunan University of Chinese Medicine, Changsha, Hunan 410208, China）

〔Abstract〕Objective To observe the protective effect of modified Danshen Decoction containing serum on p38MARK expression in myocardial cells of hypoxia/reoxygenation（H/R）neonatal rats and ivvestigate its underlying molecular mechanisms. Methods Cardiomyocytes from 20 neonatal SD rats were primary cultured, and the built H/R models were randomly divided into H/R group, SB203580 group（p38MAPK inhibitor group）, modified Danshen Decoction-containing serum group, and the normal cultured cardiomyocytes were as the control group. Cardiomyocytes were identified by T cell chemical staining method, and the morphologic changes in cardiomyocytes were observed by the Rhodamine B staining. Cytotoxicity of drug-contained sera was determined by the MTT method. Expression of MAP Kinase Kinase 3（MKK3）, MKK6, p38MAPK, and phospho-p38MAPK were measured by the Western blotting. Results Compared with the normal serum control group, the expressions of MKK3 and MKK6 were increased in H/R group, which was not altered by SB203580 and modified Danshen Decoction; The levels of p38MAPK and phospho -p38MAPK at H/R were increased, while the p38MAPK and phosphop -38MAPK expression in SB203580 and modified Danshen Decoction groups were decreased. Conclusion The modified Danshen Decoction could protectcardiomyocytes and decrease H/R -induced myocardial cells injury by inhibiting the expression and phosphorylation of p38MAPK through the inhibition of its p38MAPK signaling pathway .

〔Keywords〕modified Danshen Decocotion; cardiomyocytes; hypoxia/reoxygenation；p38MAPK signaling pathway；MAP Kinase Kinase 3; MAP Kinase Kinase 6; Salvia miltiorrhiza; sandalwood; red peony root

心肌缺血再灌注（ischemia reperfusion, IR）损伤是目前心血管疾病研究的热点，新近研究显示了中医药预处理对心肌IR损伤保护的优势[1]。p38丝裂原激活的蛋白激酶（p38MAPK）信号通路是1993年发现的一类MAPK信号通路，它通过转录因子磷酸化而改变基因的表达水平，参与多种胞内信息传递过程，被认为是细胞众多信号转导通路的中转站。……