阿司匹林抑制脂多糖诱导RAW264.7细胞MMP-9表达及机制研究

中国临床药理学与治疗学 ›› 2015, Vol. 20 ›› Issue (4): 388-393.

阿司匹林抑制脂多糖诱导RAW264.7细胞MMP-9表达及机制研究

张平¹, 沈陈林¹, 李琳¹, 张炜¹, 姚成增², 黄晓晖¹

¹安徽医科大学药学院,合肥 230032,安徽;
²上海中医药大学附属曙光医院心内科,上海 201203

收稿日期:2014-10-10 修回日期:2015-03-22 发布日期:2015-05-07
通讯作者: 黄晓晖,男,博士,教授,硕士生导师,研究方向:定量药理学和临床药代动力学。Tel: 13855183138 E-mail: mathdrug@sina.com
作者简介:张平,女,在读硕士,研究方向:临床药物代谢动力学。Tel: 15056925212 E-mail: 1063374460@qq.com
基金资助:
国家自然科学基金项目(81173133)

Inhibitory effect of aspirin on the expression of MMP-9 and its mechanism in LPS-induced RAW264.7 cells

ZHANG Ping¹, SHEN Chen-lin¹, LI Lin¹, ZHANG Wei¹, YAO Cheng-zeng², HUANG Xiao-hui¹

¹ Department of Basic and Clinical Pharmacology, School of Pharmacy, Anhui Medical University, Hefei 230032, Anhui,China;
² Department of Cardiology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China

Received:2014-10-10 Revised:2015-03-22 Published:2015-05-07

摘要/Abstract

摘要： 目的: 探讨阿司匹林对脂多糖(LPS)诱导RAW264.7细胞中基质金属蛋白酶-9(matrix metalloproteinase-9, MMP-9)的表达及其机制研究。方法: MTT法检测LPS诱导RAW264.7细胞作用下,阿司匹林对细胞毒性的影响。Q-RT- PCR检测MMP-9 mRNA表达,免疫蛋白印迹法(western blot)检测MMP-9、p38MAPK及磷酸化p38(Phospho-p38,P-p38)蛋白表达。特异性抑制剂SB203580(SB)阻断p38MAPK通路后,分别应用Q-RT-PCR和Western blot检测MMP-9的基因和蛋白表达。结果: 阿司匹林呈剂量依赖性抑制MMP-9基因和蛋白表达。与LPS组相比,阿司匹林可明显抑制P-p38MAPK蛋白的表达,而p38MAPK总蛋白无明显变化。抑制p38MAPK通路后,MMP-9 mRNA和蛋白水平均明显下调。结论: 阿司匹林抑制LPS诱导RAW264.7细胞中MMP-9表达可能与抑制p38MAPK信号转导通路有关,进而发挥其抗AS药理作用。

关键词: 阿司匹林, 基质金属蛋白酶, 动脉粥样硬化, 巨噬细胞

Abstract: AIM: To investigate the effect of aspirin on the expression of MMP-9 and its mechanism in LPS-induced RAW264.7 cells.METHODS: The cytotoxic effect of aspirin on LPS-induced RAW264.7 cells was detected by MTT assay. MMP-9mRNA expression was detected by quantitative real-time PCR. The protein expression level of MMP-9, p38MAPK and Phospho-p38 (P-p38) were examined by western blotting. After the inhibition of p38MAPK pathway by specific inhibitor SB203580, the gene and protein expression of MMP-9 was measured by Quantitative real-time PCR and western blotting separately.RESULTS: The gene and protein expression of MMP-9 were inhibited by aspirin in dose-dependent manner.The protein expression of P-p38 was significantly inhibited by aspirin as compared with LPS treatment group. However, the protein expression of p38MAPK was not significantly different. The expression of MMP-9mRNA and protein were obviously decreased after inhibiting p38MAPK pathway.CONCLUSION: The results suggest that aspirin has potent anti-atherosclerotic action with inhibitory action on MMP-9 production by blocking p38MAPK pathway in LPS-induced RAW264.7 cells.

Key words: aspirin, matrix metalloproteinase, atherosclerosis, macrophage

中图分类号:

R965.2

张平, 沈陈林, 李琳, 张炜, 姚成增, 黄晓晖. 阿司匹林抑制脂多糖诱导RAW264.7细胞MMP-9表达及机制研究[J]. 中国临床药理学与治疗学, 2015, 20(4): 388-393.

ZHANG Ping, SHEN Chen-lin, LI Lin, ZHANG Wei, YAO Cheng-zeng, HUANG Xiao-hui. Inhibitory effect of aspirin on the expression of MMP-9 and its mechanism in LPS-induced RAW264.7 cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2015, 20(4): 388-393.

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