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培养的肥厚心肌细胞中RhoA/ROCK通路和ERK通路的相互对话

发表时间:2016-03-15  浏览量:1295  下载量:440
全部作者: 李乐,张可,陶厚权,尚好,张彩玲
作者单位: 浙江工业大学药学院;浙江省人民医院中心实验室;浙江人才学院
摘 要: 目的:探讨心肌肥厚(cardiac hypertrophy,CH)心肌细胞中RhoA(ras homologue A)/ROCK(Rho相关激酶)通路和细胞外信号调节蛋白激酶(extra cellular signal regulated protein kinase,ERK)通路是否存在相互作用。方法:原代培养的大鼠乳鼠心肌细胞,分为四组:对照组、RhoA组、ERK阻断剂(PD98059)组和RhoA+ERK阻断剂(PD98059)组。分别采用Western blotting法检测CH心肌细胞中ROCK、ERK1/2蛋白表达及磷酸化ROCK(p-ROCK)、磷酸化ERK1/2(p-ERK1/2)蛋白含量,逆转录PCR(reverse transcription PCR,RT-PCR)方法测CH心肌细胞中ROCK蛋白的表达。结果:与对照组相比,RhoA组p-ROCK、p-ERK1/2蛋白含量增多(P<0.05),ERK阻断剂组p-ROCK蛋白含量无差异、p-ERK1/2蛋白含量减少(P<0.05),RhoA+ERK阻断剂组p-ROCK、p-ERK1/2蛋白含量无差异;与RhoA组相比,RhoA+ERK阻断剂组p-ROCK、p-ERK1/2蛋白含量减少(P<0.05)。各组间ROCK、ERK1/2蛋白表达无差异。各组的ROCK mRNA表达差异显著。结论:RhoA可直接活化ROCK蛋白和ERK1/2通路,ERK1/2通路可能位于RhoA/ROCK通路的下游,ROCK蛋白的活化作用可能部分依赖于ERK1/2通路的激活。
关 键 词: 药理学;心肌肥厚细胞;RhoA;ERK
Title: Interaction between RhoA/ROCK pathway and ERK pathway in cultured hypertrophy cardiomyocyts
Author: LI Le, ZHANG Ke, TAO Houquan, SHANG Hao, ZHANG Cailing
Organization: College of Pharmaceutical Science, Zhejiang University of Technology; Central Lab, Zhejiang Province People’s Hospital; Rencai College of Zhejiang
Abstract: Objective: To investigate whether the interaction between RhoA (ras homologue A)/ROCK pathway and extra cellular signal regulated protein kinase (ERK) pathway in the cultured hypertrophy cardiomyocyts. Methods: The hearts of neonatal rats were isolated and cultured in vitro,then the cardiomyocyts were divided into four groups which stimulated by same dosage isoprenaline respectively: control group, RhoA group, ERK blocking drug (PD98059) group, RhoA+ERK blocking drug (PD98059) group. The expression of ROCK, ERK1/2 proteins, the content of phosphorylated ERK1/2 (p-ERK1/2) and ROCK proteins (p-ROCK) were detected by Western blotting,and the expression of ROCK mRNA in the cultured hypertrophy cardiomyocyts was detected by reverse transcription PCR (RT-PCR). Results: Compared with the control group, the contents of p-ROCK and p-ERK1/2 proteins in RhoA group were increased (P<0.05), while those in ERK blocking drug group were increased (P<0.05). There was no difference between control group and RhoA+ERK blocking drug group. Compared with RhoA group, the contents of p-ROCK and p-ERK1/2 proteins in ERK blocking agent group and RhoA+ERK blocking drug group were decreased (P<0.05). There was no difference in the expression of ROCK and ERK1/2 proteins among different groups. There were obvious differences in expression of ROCK mRNA among different groups. Conclusion: RhoA may directly activate ROCK proteins and ERK1/2 pathway, and the ERK1/2 pathway may be located in the downstream of the RhoA/ROCK pathway. The activation of ROCK proteins may partially depend on ERK1/2 pathway stimulation.
Key words: pharmacology; cardiac hypertrophy cell; RhoA; ERK
发表期数: 2016年3月第5期
引用格式: 李乐,张可,陶厚权,等. 培养的肥厚心肌细胞中RhoA/ROCK通路和ERK通路的相互对话[J]. 中国科技论文在线精品论文,2016,9(5):470-475.
 
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