23-羟基白桦酸对阿霉素所致H9c2心肌细胞损伤的保护作用及抗氧化机制研究

中国临床药理学与治疗学 ›› 2012, Vol. 17 ›› Issue (7): 736-743.

23-羟基白桦酸对阿霉素所致H9c2心肌细胞损伤的保护作用及抗氧化机制研究

郝刚¹, 周芳¹, 刘嘉莉¹, 王广基¹, 桑国卫², 叶文才³

¹中国药科大学药物代谢动力学重点实验室,南京 210009,江苏;
²中国药品生物制品检定所,北京 100050;
³暨南大学中药及天然药物研究所,广州 510632,广东

收稿日期:2012-03-20 修回日期:2012-05-25 发布日期:2012-07-17
通讯作者: 王广基,男,博士,教授,博士生导师,研究方向:药物代谢动力学。Tel: 025-83271182 E-mail: guangjiwang@hotmail.com
桑国卫,男,博士,中国工程院院士,研究方向:临床药理。Tel: 010-67095371 E-mail: sanggw@nicpbp.org.cn
作者简介:郝刚,男,博士研究生,研究方向:药物代谢动力学。Tel: 025-83271176 E-mail: haogangnice@yahoo.com.cn
基金资助:
“十一五”重大新药创制专项资助(2009ZX09304-001);国家自然科学基金资助(30801411);中央高校基本科研业务费专项资金资助 (JKY2009012)

Protective effect of 23-hydroxybetulinic acid against doxorubicin-induced cardiotoxicity through the antioxidant activity in H9c2 cells

HAO Gang¹, ZHOU Fang¹, LIU Jia-li¹, WANG Guang-ji¹, SANG Guo-wei², YE Wen-cai³

¹Key Laboratory of Drug Metabolism and Pharmacokinetics, Nanjing 210009, Jiangsu, China;
²National Institute for the Control of Pharmaceutical and Biological Products, Beijing 100050, China;
³Institute of Traditional Chinese Medicine and Nature Products, Jinan University, Guangzhou 510632, Guangdong, China

Received:2012-03-20 Revised:2012-05-25 Published:2012-07-17

摘要/Abstract

摘要： 目的: 基于细胞水平研究23-羟基白桦酸(23-HBA)对阿霉素(DOX)心肌损伤的保护作用并探讨其抗氧化机制。 方法: 采用H9c2心肌细胞,考察对照组、DOX组、23-HBA组及23-HBA联合DOX组给药后心肌细胞形态学变化,测定细胞体积及蛋白含量,Real time-PCR测定心钠素(ANP)、脑钠素(BNP)的mRNA水平。考察心肌细胞内Caspase-3活性及Hoechst荧光染色后细胞核形态从而评价细胞凋亡水平。利用DCFH荧光探针测定细胞ROS水平,同时检测细胞内脂质过氧化物(MDA)生成量,评价细胞氧化损伤水平,寻找23-HBA心肌保护的可能机制。 结果: 5 μmol/L DOX诱导心肌细胞肥大,表现为细胞体积增大,蛋白含量增多,并显著上调细胞内ANP与BNP的mRNA水平,提高细胞内Caspase-3活性,导致细胞凋亡。23-HBA与DOX联合用药后,浓度依赖地降低DOX给药后心肌细胞内ROS水平,清除脂质过氧化物MDA,逆转DOX诱导的心肌肥大、心肌损伤标志物上调、心肌细胞凋亡等过程。 结论: 23-HBA可通过其抗氧化作用降低DOX所致的心肌损伤。

关键词: 23-羟基白桦酸, 阿霉素, 心肌损伤

Abstract: AIM: To investigate the potential protective effects of 23-hydroxybetulinic acid (23-HBA) on doxorubicin (DOX)-induced cardiotoxicity and the underlying mechanisms. METHODS: H9c2 cells were treated with DOX, 23-HBA, or their combinations, cell size and protein content were determined to evaluate the cardiac hypertrophy induced by DOX. Expression of ANP and BNP mRNA were studied by Real-time PCR. DOX-induced caspase-3 activation and apoptosis were further studied. Oxidative stress and lipid peroxidation were determined to evaluate the potential mechanism of the cardioprotective effect of 23-HBA. RESULTS: 23-HBA remarkably reduced the mRNA level of ANP and BNP induced by DOX, and reversed the cardiac hypertrophy caused by DOX treatment. Caspase-3 activity also significantly reduced when combined treated with 23-HBA and DOX compared to DOX treatment. Further studies showed that 23-HBA exerted its cardioprotective effect on DOX through decreasing the intracellular ROS and MDA amount. CONCLUSION: 23-HBA protected H9c2 cells against the cardiotoxicity of DOX, and the reduction of oxygen free radicals in cardiac myocytes may be the underlying mechanism mediating the protective effect of 23-HBA.

Key words: 23-HBA, Doxorubicin, Cardiotoxicity

中图分类号:

R965.2

郝刚, 周芳, 刘嘉莉, 王广基, 桑国卫, 叶文才. 23-羟基白桦酸对阿霉素所致H9c2心肌细胞损伤的保护作用及抗氧化机制研究[J]. 中国临床药理学与治疗学, 2012, 17(7): 736-743.

HAO Gang, ZHOU Fang, LIU Jia-li, WANG Guang-ji, SANG Guo-wei, YE Wen-cai. Protective effect of 23-hydroxybetulinic acid against doxorubicin-induced cardiotoxicity through the antioxidant activity in H9c2 cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(7): 736-743.

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