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阿利克仑抑制血管紧张素原-肾素双转基因高血压小鼠心肌成纤维细胞增殖

王丽萍^*, 范素净, 李树民, 王小君, 孙娜

华北理工大学基础医学院生理学系；唐山市慢性病临床基础研究重点实验室，唐山 063000

摘要

本研究旨在观察阿利克仑(aliskiren)在血管紧张素原-肾素(AGT-REN)双转基因高血压(double transgenic hypertension, dTH)小鼠心肌成纤维细胞(cardiac fibroblasts, CFs)增殖中的作用。将培养的AGT-REN dTH小鼠CFs分为高血压组(dTH)和阿利克仑干预组；另培养同品系野生C57B6小鼠CFs作为对照(WT)。采用MTT法观察不同浓度阿利克仑(1 × 10⁻⁶、1 × 10⁻⁷、1 × 10⁻⁸、1 × 10⁻⁹ mol/L)对dTH小鼠CFs增殖的影响；选取1 × 10⁻⁷ mol/L阿利克仑作用dTH小鼠CFs 24 h，羟脯氨酸试剂盒定量检测细胞胶原含量；Western blot法检测细胞I、III型胶原蛋白表达，观察细胞胶原合成变化；Western blot法检测α-SMA表达，观察细胞转化；DHE检测细胞内活性氧表达，Western blot法检测细胞内NADPH氧化酶蛋白表达，观察细胞内氧化应激反应的改变。结果显示，随年龄增长，AGT-REN dTH组小鼠血压及血浆Ang II水平均呈明显升高趋势，与WT组小鼠相比，CFs增殖明显。1 × 10−6、1 × 10−7、1 × 10−8 mol/L阿利克仑均能抑制AGT-REN dTH小鼠CFs增殖。1 × 10−7 mol/L阿利克仑使dTH小鼠CFs表达α-SMA减少，胶原合成及I、III型胶原蛋白表达下降，活性氧表达减少；同时，阿利克仑降低了dTH小鼠CFs内NADPH氧化酶NOX2及NOX4的蛋白表达。阿利克仑抑制AGT-REN dTH小鼠CFs增殖、表型转化及胶原合成，这一过程可能与其抑制细胞内氧化应激反应有关。

关键词： 阿利克仑; 心肌成纤维细胞; 氧化应激; 心肌纤维化

分类号：R542.2

[Aliskiren inhibits proliferation of cardiac fibroblasts in AGT-REN double transgenic hypertensive mice in vitro.] [Article in Chinese]

WANG Li-Ping^*, FAN Su-Jing, LI Shu-Min, WANG Xiao-Jun, SUN Na

Department of Physiology； Tangshan Key Laboratory for Preclinical on Chronic Diseases, School of Basic Medicine, North China University of Science and Technology, Tangshan 063000, China

Abstract

The purpose of the present study is to explore the effect of aliskiren on the proliferation of cardiac fibroblasts (CFs) in AGT-REN double transgenic hypertensive (dTH) mice. The cultured CFs from AGT-REN dTH mice were divided into AGT-REN group (dTH) and aliskiren group (ALIS). Cultured CFs from C57B6 mice were served as control (WT). The effect of different concentration of aliskiren (1 × 10⁻⁶, 1 × 10⁻⁷, 1 × 10⁻⁸, 1 × 10⁻⁹ mol/L) on CFs proliferation was determined by MTT assay. After treatment with 1 × 10⁻⁷ mol/L aliskiren for 24 h, α-SMA, collagen I, III and NADPH oxidase (NOX) protein expression in CFs of AGT-REN dTH mice were detected by Western blot. The collagen synthesis in CFs was assessed by hydroxyproline kit. The expression of ROS was determined by DHE. Results showed that the blood pressure and plasma Ang II levels were significantly increased and CFs proliferation was significantly increased as well in AGT-REN dTH mice compared with WT group. However, aliskiren intervention decreased CFs proliferation, myofibroblast transformation, as well as the collagen I and III synthesis in CFs of AGT-REN dTH mice. Meanwhile, aliskiren inhibited ROS content and NOX2/NOX4 protein expression in CFs of AGT-REN dTH mice. These results suggest that aliskiren decreases the cell proliferation, myofibroblast transformation and collagen production in CFs of AGT-REN dTH mice, which might be through inhibition of oxidative stress response.

Key words: aliskiren; cardiac fibroblast; oxidative stress; myocardial fibrosis

收稿日期：2016-01-05　　录用日期：2016-06-29

通讯作者：王丽萍　　E-mail: mywlpzjm@163.com

引用本文：

王丽萍, 范素净, 李树民, 王小君, 孙娜. 阿利克仑抑制血管紧张素原-肾素双转基因高血压小鼠心肌成纤维细胞增殖[J]. 生理学报 2016; 68 (5): 684-690.

WANG Li-Ping, FAN Su-Jing, LI Shu-Min, WANG Xiao-Jun, SUN Na. [Aliskiren inhibits proliferation of cardiac fibroblasts in AGT-REN double transgenic hypertensive mice in vitro.] [Article in Chinese]. Acta Physiol Sin 2016; 68 (5): 684-690 (in Chinese with English abstract).