Pyk2促进血管紧张素II诱导的大鼠心肌细胞肥大

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (3): 351-355.

Pyk2促进血管紧张素II诱导的大鼠心肌细胞肥大

鄢雯, 刘立新, 孔祥照, 骆雨辰, 李中秋, 李囡^*

首都医科大学燕京医学院, 北京 101300

收稿日期:2019-05-31 修回日期:2019-11-04 出版日期:2020-03-05 发布日期:2020-03-02
通讯作者: ^*linan881219@126.com
基金资助:
首都医科大学燕京医学院自然科学基金(2016QD04)

Pyk2 promotes Ang Ⅱ-induced cardiac hypertrophy in rat

YAN Wen, LIU Li-xin, KONG Xiang-zhao, LUO Yu-chen, LI Zhong-qiu, LI Nan^*

Yanjing Medical College,Capital Medical University,Beijing 101300

Received:2019-05-31 Revised:2019-11-04 Online:2020-03-05 Published:2020-03-02
Contact: ^*linan881219@126.com

摘要/Abstract

摘要： 目的探讨富含脯氨酸蛋白酪氨酸激酶(Pyk2)对血管紧张素Ⅱ(Ang Ⅱ)诱导大鼠心肌细胞肥大的作用及其机制。方法提取大鼠乳鼠原代心肌细胞,随机分为对照组、Ang Ⅱ诱导组(Ang Ⅱ剂量为100 nmol/L,将其加入心肌细胞培养皿中刺激24 h)、Pyk2抑制剂PF-431396组(PF-431396剂量为10 μmol/L,在Ang Ⅱ刺激前30 min加入心肌细胞培养皿中)以及PF-431396干预Ang Ⅱ诱导组(Ang Ⅱ剂量为100 nmol/L,PF-431396剂量为10 μmol/L)。用细胞骨架蛋白(α-actinin)免疫荧光染色检测心肌细胞表面积大小;real-time PCR检测心肌细胞中肥大指标(ANP和β-MHC)mRNA表达;Western blot检测心肌细胞p-Pyk2、Pyk2以及p53的蛋白表达。结果在Ang Ⅱ刺激下,心肌细胞表面积增大(P＜0.05)、肥大指标的mRNA水平升高(P＜0.05)、p-Pyk2和p53的蛋白水平增加(P＜0.05);而加入Pyk2抑制剂PF-431396后,心肌肥大指标明显改善,p-Pyk2和p53的蛋白表达降低(P＜0.05)。结论 Pyk2通过p53信号通路促进Ang Ⅱ诱导的大鼠心肌细胞肥大。

关键词: 心肌肥大, 血管紧张素Ⅱ(AngⅡ), 富脯氨酸蛋白酪氨酸激酶(Pyk2)

Abstract: Objective To investigate the role of proline-rich protein tyrosine kinase (Pyk2) in angiotensin Ⅱ(Ang Ⅱ)-induced cardiac hypertrophy and the underlying mechanism in rat. Methods Primary rat cardio-myocytes were isolated from rats and randomly divided into control group, Ang Ⅱ induction group (the dose of Ang Ⅱ was 100 nmol/L, cardiomyocytes were stimulated in the cell culture dish for 24 hours), Pyk2 inhibitor PF-431396 group (the dose of PF-431396 was 10 μmol/L, added to the cell culture dish 30 minutes in advance before Ang Ⅱ stimulation) and Ang Ⅱ induction group intervened by PF-431396 (the dose of Ang Ⅱ was 100 nmol/L, the dose of PF-431396 was 10 μmol/L). The size of cardiomyocytes was measured by α-actinin fluorescence staining. RT-qPCR was performed to detect the mRNA level of hypertrophic markers (ANP and β-MHC) in cardiomyocytes. The expressions of p-Pyk2, Pyk2 and p53 in cardiomyocytes were detected by Western blot analysis. Results Ang Ⅱ stimulation increased the surface area of cardiomyocytes and the mRNA level of hypertrophic markers(P＜0.05). In addition, the proteins of p-Pyk2 and p53 were increased(P＜0.05). However, after treatment of Pyk2 inhibitor PF-431396, moycardial hypertrophy was significantly improved and the protein level of Pyk2 and p53 decreased(P＜0.05). Conclusions Pyk2 promotes cardiomyocyte hypertrophy in rat induced by Ang Ⅱ through the p53 signaling pathway.

Key words: cardiac hypertrophy, angiotensin Ⅱ(Ang Ⅱ), proline-rich tyrosine kinase 2 (Pyk2)

中图分类号:

鄢雯, 刘立新, 孔祥照, 骆雨辰, 李中秋, 李囡. Pyk2促进血管紧张素II诱导的大鼠心肌细胞肥大[J]. 基础医学与临床, 2020, 40(3): 351-355.

YAN Wen, LIU Li-xin, KONG Xiang-zhao, LUO Yu-chen, LI Zhong-qiu, LI Nan. Pyk2 promotes Ang Ⅱ-induced cardiac hypertrophy in rat[J]. Basic & Clinical Medicine, 2020, 40(3): 351-355.

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