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尿促皮素诱导乳大鼠心肌细胞肥大的作用及信号传导机制
引用本文:梁春光,王洪新,黄雷,刘杰. 尿促皮素诱导乳大鼠心肌细胞肥大的作用及信号传导机制[J]. 中国药理学与毒理学杂志, 2009, 23(6): 436-442. DOI: 10.3867/j.issn.10003002.2009.06.004
作者姓名:梁春光  王洪新  黄雷  刘杰
作者单位:1. 辽宁医学院药理学教研室,辽宁,锦州,121001
2. 锦州市药品检验所,辽宁,锦州,121001
基金项目:辽宁省高等学校优秀人才支持计划资助项目,辽宁省教育厅创新团队资助项目 
摘    要:目的探讨尿促皮素(urocortin)诱导大鼠心肌细胞肥大的作用及其信号传导机制。方法实验分8组,正常对照组、尿促皮素0.1μmol.L-1组、星形孢菌素(Sta)1μmo.lL-1、H890.1μmol.L-1和维拉帕米(Ver)1μmol.L-1组及尿促皮素分别加Sta,H89和Ver组。采用体外培养的乳大鼠心肌细胞,应用尿促皮素0.1μmol.L-1诱导心肌肥大,观察Sta1μmol.L-1,H890.1μmo.lL-1和Ver1μmol.L-1的作用,进一步探讨尿促皮素0.1μmol.L-1诱导心肌肥厚的作用机制。用消化分离法及计算机图像分析系统检测心肌细胞直径;[3H]亮氨酸掺入法测定心肌细胞蛋白质的合成;用Lowry法检测心肌细胞蛋白质含量;用Western蛋白印迹法测定心房钠尿肽(ANP)表达;采用Till阳离子测定系统,以Fura-2/AM为荧光探针,观察心肌细胞[Ca2+]i瞬间变化。结果尿促皮素使心肌细胞直径、蛋白质合成、蛋白质含量和ANP表达分别增加30.9%,36.3%,35.5%和34.7%;尿促皮素+Sta组使心肌细胞直径、蛋白质合成、蛋白质含量和ANP表达分别降低了16.5%,22.1%,18.1%和21.3%;尿促皮素+H89组使心肌细胞直径、蛋白质合成、蛋白质含量和ANP表达分别降低了16.6%,21.5%,19.5%和20.6%;尿促皮素+Ver组使心肌细胞直径、蛋白质合成、蛋白质含量和ANP表达分别降低了17.1%,20.9%,17.9%及19.9%;尿促皮素能够使心肌细胞[Ca2+]i瞬间变化水平增高,Sta,H89和Ver能够降低尿促皮素引起的心肌细胞[Ca2+]i瞬间变化升高。结论尿促皮素可能通过蛋白激酶C和蛋白激酶A信号途径影响L-型Ca2+通道,进而影响细胞[Ca2+]i瞬间变化水平,诱导乳大鼠心肌细胞肥大。

关 键 词:尿促皮素  肌细胞,心脏  信号传导通路
收稿时间:2009-03-18

Effect and mechanism of signal transduction pathway in urocortininduced cardiomyocytes hypertrophy in neonatal rat
LIANG Chun-Guang,WANG Hong-Xin,HUANG Lei,LIU Jie. Effect and mechanism of signal transduction pathway in urocortininduced cardiomyocytes hypertrophy in neonatal rat[J]. Chinese Journal of Pharmacology and Toxicology, 2009, 23(6): 436-442. DOI: 10.3867/j.issn.10003002.2009.06.004
Authors:LIANG Chun-Guang  WANG Hong-Xin  HUANG Lei  LIU Jie
Affiliation:LIANG Chun-Guang1, WANG Hong-Xin1, HUANG Lei2, LIU Jie1
Abstract:AIM To investigate the effects and mechanism of cardiomyocyte hypertrophy induced by urocortin. METHODS The cardiomyocytes were divided into 8 groups: normal control, urocortin, staurosporine(Sta), verapamil(Ver), H89, urocortin+Sta, urocortin+Ver, and urocortin+H89 groups. The cardiomyocytes diameter was measured by computer photograph analysis system. The protein synthetic rate was obtained through measuring the incorporation of [~3H]-leucine into myocyte protein by liquid scintillation method. The total protein content was assayed by Lowry method. The expression of atrial natriuretic peptide (ANP) was determined by Western blot. [Ca~(2+)]_i transient was measured by Till image system by cell loading Fura-2/AM. RESULTSUrocortin group enhanced cardiomyocyte volume, protein synthesis, total protein content and expression of ANP by 30.9%, 36.3%, 35.5% and 34.7%;urocortin+Sta group decreased cardiomyocyte diameter, protein synthesis, total protein content and expression of ANP by 16.5%, 22.1%, 18.1% and 21.3%;urocortin+H89 group decreased the cardiomyocyte diameter, the protein synthesis,total protein content and expression of ANP by 16.6%, 21.5%, 19.5% and 20.6%;urocortin+Ver decreased the cardiomyocyte diameter, the protein synthesis, total protein content and the expression of ANP by 17.1%, 20.9%, 17.9% and 19.9%;Sta, H89 and Ver could decrease the [Ca~(2+)]_i transient induced by urocortin. CONCLUSION The hypertrophic effect of urocortin in rat neonatal cardiomyocytes is mediated via activation of protein kinase C and protein kinase A pathway and L-type calcium channels.
Keywords:urocortin  myocytes,cardiac  signal transduction pathway
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