Pyk2促进血管紧张素Ⅱ诱导的大鼠心肌细胞肥大  

Pyk2 promotes Ang Ⅱ-induced cardiac hypertrophy in rat

作　　者：鄢雯[1] 刘立新[1] 孔祥照[1] 骆雨辰 李中秋[1] 李囡[1] YAN Wen;LIU Li-xin;KONG Xiang-zhao;LUO Yu-chen;LI Zhong-qiu;LI Nan(Yanjing Medical College,Capital Medical University,Beijing 101300)

机构地区：[1]首都医科大学燕京医学院

出　　处：《基础医学与临床》2020年第3期351-355,共5页Basic and Clinical Medicine

基　　金：首都医科大学燕京医学院自然科学基金(2016QD04)

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(AngⅡ)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、AngⅡ诱导组(AngⅡ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10μmol/L,在AngⅡ刺激前30 min加入心肌细胞培养皿中)以及PF-431396干预AngⅡ诱导组(AngⅡ剂量为100 nmol/L,PF-431396剂量为10μmol/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AngⅡ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AngⅡ诱导的大鼠心肌细胞肥大。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 cardiomyocytes were isolated from rats and randomly divided into control group,Ang Ⅱ induction group( the dose of AngⅡ 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.

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) 

分 类 号：R33[医药卫生—人体生理学]