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陈聪,廖菁,李鑫,宋厚盼,黄政德.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[J].湖南中医药大学学报英文版,2016,36(2):35-39.[Click to copy
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| 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 |
| DOI：10.3969/j.issn.1674-070X.2016.02.010 |
| Received:September 08, 2015 |
| 基金项目:国家自然科学基金资助项目（81373576）；湖南省中医药科研计划课题（B0347）；湖南省教育厅科学研究项目（14C0872）；湖南省科技厅科学研究项目（2014SK3034）；湖南省中医药管理局重点课题（201304）；湖南中医药大学优秀教师培养计划（2015）。 |
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| 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 protect cardiomyocytes and decrease H/R-induced myocardial cells injury by inhibiting the expression and phosphorylation of p38MAPK through the inhibition of its p38MAPK signaling pathway. |
| Key words: modified Danshen Decocotion cardiomyocytes hypoxia/reoxygenation p38MAPK signaling pathway MAP Kinase Kinase 3 MAP Kinase Kinase 6 Salvia miltiorrhiza sandalwood red peony root |
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