氯沙坦对心肌成纤维细胞基质金属蛋白酶-2 、JNK1/2 表达及增殖活性的影响

中国临床药理学与治疗学 ›› 2007, Vol. 12 ›› Issue (1): 72-77.

氯沙坦对心肌成纤维细胞基质金属蛋白酶-2 、JNK1/2 表达及增殖活性的影响

肖云彬¹, 秦旭平², 覃丽², 廖端芳², 黄红林²

¹浙郴州市第一人民医院药剂科临床药学室, 郴州 423000, 湖南;
²南华大学药物药理研究所, 衡阳 421000, 湖南

收稿日期:2006-05-22 修回日期:2006-09-30 出版日期:2007-01-26 发布日期:2020-10-26

Effects of losartan on expression of matrix metalloproteinase-2,JNK1/2 and proliferation in cardiac fibroblast

XIAO Yun-bin¹, QIN Xu-ping², QIN Li², LIAO Duan-fang², HUANG Hong-lin²

¹Pharmacy Department, the First Hospital of Chenzhou, Chenzhou 423000, Hunan, China;
²Institute of Pharmacy and Pharmacology, Nanhua University, Hengyang 421001, Hunan, China

Received:2006-05-22 Revised:2006-09-30 Online:2007-01-26 Published:2020-10-26
Contact: HUANG Hong-Lin, female, professor, tutor of master,engaged in protective efficacy of cardiovascular system of estrogen.Tel:0735-8281408 E-mail: huanghonglinhui@yahoo.com.cn
About author:XIAO Yun-bin, female, pharmacist, engaged in cardiovascular pharmacy.Tel:0735-2227769 　E-mail:xiaoyunbin.student@sina.com

摘要/Abstract

摘要： 目的: 研究氯沙坦通过影响高血压大鼠心肌成纤维细胞间质成分抑制心室重构的药理机制。方法: 培养心肌成纤维细胞, 免疫细胞化学法鉴定细胞,MTT 法测氯沙坦和Ang Ⅱ对细胞增殖活性影响的量效关系, Western Blotting 测Ang Ⅱ刺激成纤维细胞心肌JNK1/2 、磷酸化JNK1/2 表达的时效关系。根据实验所得Ang Ⅱ刺激心肌成纤维细胞增殖活性作用的EC₅₀ 值、氯沙坦抑制心肌成纤维细胞活性作用的IC₅₀值和Ang Ⅱ刺激JNK1/2 表达的最佳时间, 心肌成纤维细胞分为无血清DMEM 组、Ang Ⅱ100 nmol/L组、Ang Ⅱ 100 nmol/L +losartan 100 nmol/L组、losartan 100 nmol/L组, 药物干预后分别收集各组蛋白质、培养上清液,Western blotting 检测各组MMP-2 、JNK1/2 、磷酸化JNK1/2 蛋白表达, ELISA 检测分泌至培养上清液中MMP-2 的量。结果: Ang Ⅱ 刺激心肌成纤维细胞增殖活性作用的EC50为53 nmol/L, 氯沙坦抑制心肌成纤维细胞增殖活性作用的EC₅₀ 为56.3 nmol/L 。JNK1 2 蛋白表达在Ang Ⅱ刺激2 min达高峰, 随后表达逐渐降低。Ang Ⅱ增加JNK1/2 表达, 氯沙坦降低Ang Ⅱ 刺激导致的JNK1 2 表达。Ang Ⅱ组MMP-2 蛋白表达较无血清DMEM 组明显增高, 氯沙坦降低Ang Ⅱ刺激增高的MMP-2 表达。AngⅡ组MMP-2 分泌较无血清DMEM 组增高, 氯沙坦减少Ang Ⅱ刺激所致MMP-2 分泌增高。结论: 氯沙坦防治高血压引起的心室重构, 可能与其对抗AngⅡ刺激心肌成纤维细胞增殖活性、MMP-2 合成、分泌有关, 信号通路涉及JNK1 2 。

关键词: 氯沙坦, 基质金属蛋白酶-2, JNK1 2, 心肌成纤维细胞

Abstract: AIM: To elucidate the effects of losartan on the expression of matrix metalloproteinases-2, JNK1/2 and proliferation in cardiac fibroblast. METHODS: Neonatal rat cardiac fibroblasts were cultured.The cells proliferation was determined by MTT.To determine effects of Ang Ⅱ on JNK1/2 activity, cells were incubated (for 0,2, 5, 10, 30, 60, 120 min) in serum-free media with Ang Ⅱ, and the other group fibroblasts were exposed to serum-free media with or without Ang Ⅱ and losartan (Ang Ⅱ 100 nmol/L, Ang Ⅱ 100 nmol/L +losartan 100 nmol/L, losartan100 nmol/L, losartan for 45 min before).Cells protein was collected with MBST buffer.The relative abundance of MMP-2, JNK1/2 and p-JNK1/2 in cells was determined by immunoblotting.The secretion of MMP-2 in media of cell culture was determined by ELISA. RESULTS: Ang Ⅱ increased the proliferation of CFB in a dose-dependent manner, whereas losartan decreased the proliferation of CFB stimulated by Ang Ⅱ in a dose-dependant manner, too (P<0.05).The relative abundance of JNK1/2 was highest in Ang Ⅱ of the 2-minstimulated group.Ang Ⅱ increased expression of JNK1/2 andMMP-2 protein (P<0.05), on the contrary, losartan inhibited JNK1/2 and MMP-2 protein expression. CONCLUSION: Ang Ⅱ induce the increase of proliferation of CFB, expression of JNK1/2 and MMP-2 in CFB, and losartan inhibits these effects of Ang Ⅱ.

Key words: matrix metalloproteinase-2, JNK1/2, losartan, cardiac fibroblasts

中图分类号:

R969

肖云彬, 秦旭平, 覃丽, 廖端芳, 黄红林. 氯沙坦对心肌成纤维细胞基质金属蛋白酶-2 、JNK1/2 表达及增殖活性的影响[J]. 中国临床药理学与治疗学, 2007, 12(1): 72-77.

XIAO Yun-bin, QIN Xu-ping, QIN Li, LIAO Duan-fang, HUANG Hong-lin. Effects of losartan on expression of matrix metalloproteinase-2,JNK1/2 and proliferation in cardiac fibroblast[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2007, 12(1): 72-77.

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