心肌梗死大鼠钙敏感受体表达与心肌细胞凋亡的变化

doi:10.12047/j.cjap.5753.2019.056

摘要/Abstract

摘要： 目的:观察大鼠急性心肌梗死后不同时间心肌钙敏感受体(CaSR)的表达和心肌细胞凋亡的变化情况。方法:健康Wistar大鼠随机分为假手术组(Sham)和心肌梗死(AMI)组,通过结扎左侧冠状动脉前降支的方法,建立大鼠心肌梗死模型,分别在手术后1、2、4周(每组成功存活n=5)检测心脏形态学和血流动力学的改变,检测心肌组织中CaSR mRNA和蛋白的表达,以及Bax、Bcl- 2、caspase-3和caspase-9蛋白的表达,检测血清中乳酸脱氢酶( LDH ) 、肌酸激酶( CK )活性和肌钙蛋白( cTnT) 水平,观察心肌细胞凋亡情况。结果:和Sham组相比,随着心肌梗死的发展,AMI组大鼠心肌组织CaSR的mRNA和蛋白的表达、细胞凋亡指数均明显增加(P＜0.05)),心肌细胞超微结构损伤严重;左心室收缩压(LVSP)、左心室内压最大上升速率(+dp/dt_max)(mmHg/s)和最大下降速率 (-dp/dt_max)(mmHg/s)减少,左心室舒张末期压(LVEDP)明显增大(P＜0.05);AMI组血清cTnT 水平、CK和LDH活性均升高 (P＜0.05),随着心肌梗死的发展,cTnT 水平和CK活性逐渐降低,LDH变化不明显。心肌组织中促凋亡相关蛋白Bax、caspase-3、caspase-9表达增多,抑制凋亡的相关蛋白(或因子)Bcl-2表达减少(P＜0.05)。结论:随着AMI的发展,AMI组大鼠心肌组织中CaSR的mRNA和蛋白的表达增多,细胞凋亡数增加,表明CaSR参与了心肌梗死的发展,其机制可能与促进细胞凋亡有关。

关键词: 钙敏感受体, 急性心肌梗死, 细胞凋亡, 大鼠

Abstract: Objective: To investigate the change of calcium sensing receptor (CaSR) expression at different time in rat tissue with acute myocardial infarction (AMI) and its effect on cardiomyocyte apoptosis. Methods: The healthy Wistar rats were randomly divided into Sham and AMI groups, the rat myocardial infarction model was established by ligating left anterior descending coronary artery. The changes of cardiac morphology and hemodynamics were detected at 1, 2 and 4 weeks,respectively. The expressions of CaSR mRNA and protein in myocardial tissue were detected by RT-PCR and Western blot, respectively. The expressions of Bax, Bcl-2, caspase-3 and caspase-9 proteins were detected by Western blot. The serum levels of lactate dehydrogenase (LDH), creatine kinase (CK) activity and cardiac troponin (cTnT) were determined. The apoptosis of cardiomyocytes were tested by TUNEL staining. Results: Compared with the sham group, the expressions of CaSR mRNA and protein, the apoptosis index were increased significantly with the development of AMI (P＜0.05). The ultrastructural damage of cardiomyocytes was serious; the levels of LVSP, +dp/dt_max and -dp/dt_max were decreased,while the levels of LVEDP was increased (P＜0.05); In AMI group, the cTnT level, CK and LDH activities were all increased (P＜0.05). With the development of myocardial infarction, the cTnT level and CK activity were gradually decreased, while the activity of LDH was not significantly changed. The expressions of promote apoptosis-related Bax, caspase-3 and caspase-9 were significantly increased, and the expression of inhibited apoptosis-related protein(factor)Bcl-2 was significantly decreased (P＜0.05). Conclusion: With the development of myocardial infarction,the expressions of CaSR mRNA and protein,the apoptosis index in rat myocardial tissue were increased with time prolongation after AMI. The increased expression of CaSR is involved in rat myocardial infarction, which is related with apoptosis.

Key words: calcium sensing receptor, acute myocardial infarction, cells apoptosis, rat

袁辉, 扬国宏, 李姝, 李丽, 宋高臣, 徐长庆, 孙健. 心肌梗死大鼠钙敏感受体表达与心肌细胞凋亡的变化[J]. 中国应用生理学杂志, 2019, 35(3): 268-272.

YUAN Hui, YANG Guo-hong, LI Shu, LI Li, SONG Gao-chen, XU Chang-qing, SUN Jian. The expression of calcium-sensing receptor in rats with acute myocardial infarction and its effect on cells apoptosis[J]. CJAP, 2019, 35(3): 268-272.

参考文献

[1] Sun YH, Liu MN, Li H, et al. Calcium-sensing receptor induces rat neonatal ventricular cardiomyocyte apoptosis[J]. Biochem Biophys Res Commun, 2006, 350 (4): 942- 948.
[2] Wang LN, Wang C, Lin Y, et al. Involvement of calcium-sensing receptor in cardiac hypertrophy-induced by angiotensin II through calcineurin pathway in cultured neonatal rat cardiomyocytes[J]. Biochem Biophys Res Commun, 2008, 369(2) : 584- 589.
[3] Wang R, Xu C, Zhao W, et al. Calcium and polyamine regulated calcium-sensing receptors in cardiac tissues[J]. Eur J Biochem, 2003, 270(12): 2680-2688.
[4] Zhang WH, Fu SB, Lu FH, et al. Involvement of calcium-sensing receptor in ischemia/reperfusion-induced apoptosis in rat cardiomyocytes[J]. Biochem Biophys Res Commun, 2006, 347(4): 872-881.
[5] ChouSH, Lin SZ, Kuo WW, et al. Mesenchymal stem cell insights: prospects in cardiovascular therapy[J]. Cell Transplant, 2014, 23(4-5): 513-529.
[6] 贾振, 孙建, 李鸿珠, 等. 钙敏感受体对大鼠糖尿病性心肌病的影响[J]. 中国应用生理学杂志, 2015, 31(1): 35-37.
[7] Guo J, Li HZ, Wang LC, et al. Increased expression of calcium-sensing receptors in atherosclerosis confers hypersensitivity to acute myocardial infarction in rats[J]. Mol Cell Biochem, 2012, 366(1-2): 345-354.
[8] Levine GN, Bates ER, Blankenship JC, et al. 2015 ACC/AHA/SCAI focused update on primary percutaneous coronary intervention for patients with ST-elevation myocardial infarction: An update of the 2011 ACCF/AHA/SCAI guideline for percutaneous coronary intervention and the 2013 ACCF/AHA guideline for the management of ST-elevation myocardial infarction[J]. J Am Coll Cardiol, 2016, 67(10): 1235-1250.
[9] Gottlieb RA, Burleson KO, Kloner RA, et al. Reperfusion injury induces apoptosis in rabbit cardiomyocytes[J]. J Clin Invest, 1994, 94(4): 1621-1628.
[10]孙晓娟, 冯武龙, 侯娜, 等. JAK2/STAT3 信号通路在运动预适应抗心肌细胞凋亡中的作用[J]. 中国应用生理学杂志, 2017, 33(5): 393-397.
[11]Hengartner MO.The biochemistry of apoptosis[J]. Nature, 2000, 407 (6805): 770-776.
[12]Jaattela M. Tschopp J. Caspase-independent cell death in T lymphocytes [J]. Nat immunol, 2003, 4(5): 416-423.

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