κ-阿片受体激动对异丙肾上腺素诱导的乳大鼠心肌细胞肥厚的抑制作用被引量：4

Inhibitory effect of kappa-opioid receptor stimulation on isoprenaline-induced myocardial hypertrophy of neonatal rats

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摘要目的通过观察选择性κ-阿片受体(κ-OR)激动剂U50488H抑制异丙肾上腺素(Iso)诱导乳大鼠心肌细胞肥厚的作用及其对细胞内游离钙离子浓度([Ca2+]i)瞬变及钙调素依赖蛋白激酶Ⅱ(CaMKⅡ)表达的影响,研究κ-OR激动抑制Iso诱导的大鼠心肌细胞肥厚的信号传导机制。方法以体外培养的乳大鼠心肌细胞为模型,应用β肾上腺素受体激动剂Iso10μmol.L-1诱导心肌肥大,观察U50488H1μmo.lL-1的作用,并进一步探讨在CaMKⅡ特异性抑制剂KN930.2μmol.L-1,普萘洛尔2μmol.L-1及L-钙通道阻滞剂维拉帕米1μmol.L-1存在情况下,κ-OR的激活对心肌肥厚的作用。用Lowry法检测心肌细胞蛋白含量;消化分离法及计算机图像分析系统检测心肌细胞体积;[3H]亮氨酸掺入法测定心肌细胞蛋白的合成;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;用Western蛋白印迹法测定CaMKⅡδB表达。结果Iso10μmol.L-1使心肌细胞总蛋白含量、体积和蛋白合成明显增加,U50488H1μmol.L-1抑制Iso诱导的心肌肥大,且抑制程度与KN930.2μmol.L-1,普萘洛尔2μmol.L-1及维拉帕米1μmol.L-1相似,在KN93存在的情况下,U50488H抑制Iso诱导的心肌肥大作用增强;U50488H能降低Iso引起的心肌细胞[Ca2+]i瞬间变化升高;Iso能明显增强心肌细胞内CaMKⅡδB的表达,U50488H能降低其表达。结论κ-OR激动剂U50488H可能通过降低心肌细胞[Ca2+]i瞬间变化和减少心肌细胞内CaMKⅡδB的表达,抑制Iso诱导的乳大鼠心肌细胞肥厚。 AIM To observe the inhibitive effects and signal transduction by kappa-opioid receptor （k-OR ） stimulation on hypertrophic myocardial cells induced by isoprenaline （Iso）in neonatal rats. METHODS The total protein content was assayed by the method of Lowry. The cardiomyocytes volume was measured by computer photograph analysis system and the protein synthesis was assayed with [ 3H ] leucine incorporation method. [ Ca^2＋ ]i transient was measured by Till image system by cellloading Fura-2/AM. The expression of Ca^2＋- calmodulin dependent kinase Ⅱ （ CaMK Ⅱ ） δB was determined by Western blot. RESULTS Iso enhanced the total protein content, the cardiomyocyte volume and the protein synthesis in myocardial cells. U50488H showed the function on reducing the previous mentioned indices induced by Iso, which were similar to KN93, propranolol and verapamil. US0488H also at- tenuated the hypertrophy and the expression of CaMK Ⅱ δB induced by Iso through decreasing the [ Ca^2＋ ]i. CONCLUSION Kappa-opioid receptor stimulation can abolish the hypertrophic response induced by Iso, which is partially via attenuating the augment of [ Ca^2 ＋ ] i and the high expression of CaMK Ⅱ δB induced by Iso.

作者吴国强王洪新荆黎

机构地区辽宁医学院分子生物学与新药开发重点实验室

出处《中国药理学与毒理学杂志》 CAS CSCD 北大核心 2008年第3期186-192,共7页 Chinese Journal of Pharmacology and Toxicology

基金辽宁省自然科学基金项目(20042170)~~

关键词受体阿片 k 心肌肥大 U50488H Ca^2+-钙调蛋白依赖性蛋白激酶 receptors, opioid, kappa myocardial hypertrophy U50488H Ca^2 ＋ -calmodu-lin dependent kinases

分类号 R96 [医药卫生—药理学]

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参考文献15

1Barron BA. Opioid peptides and the heart[J]. Cardiovasc Res, 1999, 43(1):13-16.
2裴建明,陈迈,毕辉,王跃民,朱运龙.κ-阿片受体在抑制心肌肥厚中的作用[J].心脏杂志,2002,14(6):465-468. 被引量：2
3Yu XC, Li HY, Wang HX, Wong TM. U50,488H inhibits effects of norepinephrine in rat cardiomyocytes-cross-talk between kappa-opioid and beta-adrenergic receptors[J]. J Mol Cell Cardiol, 1998, 30(2):405-413.
4Wang G, Wang H, Yang Y, Wong TM. Kappa-opioid receptor stimulation inhibits growth of neonatal rat ventricular myocytes[J]. Eur J Pharmacol, 2004, 498(1-3):53-58.
5杨彦玲,王洪新,杨玲,杨育红.κ受体激动对去甲肾上腺素诱导的心肌肥大的抑制作用[J].中国药理学通报,2002,18(1):91-94. 被引量：13
6Shan D, Wang H, Su Y, Jing Y, Wong TM. Kappa-opioid receptor stimulation inhibits cardiac hypertrophy induced by β1-adrenoceptor stimulation in the rat[J]. Eur J Pharmacol, 2007, 555(2-3):100-105.
7ZhouQ LiuJ XiaoYB.The effect of Ca^2+ and CaMKⅡ in the cardiac hypertrophy and heart failure.心肺血管病杂志,2006,2(25):57-59.
8Passier R, Zeng H, Frey N, Naya FJ, Nicol RL, McKinsey TA, et al. CaM kinase signaling in duces cardiac hypertrophy and activates the MEF2 transcription factor in vivo[J]. J Clin Invest, 2000, 105(10):1395-1406.
9Zhu W, Zou Y, Shiojima I, Kudoh S, Aikawa R, Hayashi D, et al. Ca^2+/calmodulin-dependent kinaseⅡ and calcineurin play critical roles in endothelin-1-induced cardiomyocyte hypertrophy[J]. J Biol Chem, 2000, 275(20):15239-15245.
10Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with Folin phenol reagent[J]. J Biol Chem, 1951, 193(1):265-275.

二级参考文献12

1Ventura C, Guarnieri C, Vaona I, et al. Dynorphin gene expression and release in the myocardial cell[J]. J Biol Chem, 1994,269: 5384-5386.
2Krumins SA, Faden AL, Feuerstein G. Opiate binding in rat hearts: modulation of binding after hemorrhagic shock[J]. Biochem Biophys Res Comm, 1985,127: 120-128.
3McLaughlin PJ. Regulation of DNA synthesis of myocardial and epicardial cells in developing rat heart by[Met5]enkephalin[J]. Am J Physiol, 1996,271: R122- R129.
4Pepe S, Xiao RP, Hohl C, et al. `Cross Talk' between opioid peptide and adrenergic receptor signaling in isolated rat heart[J]. Circulation, 1997, 95: 2122-2129.
5Yu XC, Wang HX, Zhang WM, et al. Cross-talk between cardiac κ-opioid and β-adrenergic receptors in developing hypertensive rats[J]. J Mol Cell Cardiol, 1999, 31: 597-605.
6Sandoshima J, Isumo S. The cellular and molecular response of cardiac myocyte to mechanical stress[J]. Annu Rev Physiol, 1997, 59:551-571.
7Pei JM, Zhou JJ, Bian JS, et al. Impaired[Ca2+]i and pHi responses to κ-opioid receptor stimulation in the heart of chronically hypoxic rats[J]. Am J Physiol (Cell Physiol), 2000, 279: C1483-C1494.
8Pei JM, Wang YM, Chen M, et al. Signaling pathway mediated by κ-opioid receptor is impaired in cardiac hypertrophy[J]. Acta Pharmacologica Sinica, 2001,22(10):865-960.
9Ventura C, Spurgeon H, Lakatta EG, et al.κ and δ opioid receptor stimulation affects cardiac myocyte function and Ca2+ release from an intracellular pool in myocytes and neurons[J]. Circ Res,1992, 70: 66-81.
10Knowton KU, Michel MC, Itani M, et al. The alpha 1A-adrenergic receptor subtype mediates biochemical, molecular, and morphologic features of cultured myocardial cell hypertrophy[J]. J Biol Chem,1993, 268(21):15374-15380.