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 . ......