Effect of calcium-sensing receptor in cardiac hypertrophy induced by angiotensin II in cultured neonatal rat cardiomyocytes

Q3 Medicine

中华地方病学杂志 Pub Date : 2011-11-20 DOI:10.3760/CMA.J.ISSN.1000-4955.2011.06.013

Li-na Wang, Jin-ping Guo, Bo Wu, Xiu-xiang Gao

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Abstract

Objective To explore the roles and possible mechanism of calcium-sensing receptor(CaSR) in cell cardiac hypertrophy model using angiotensin Ⅱ (Ang Ⅱ ).Methods The cultured neonatal rat ventricular myocytes were treated with Ang Ⅱ as cell cardiac hypertrophy model.Hypertrophic neonatal rat cardiomyocytes were treated with GdCl3(a specific agonist of CaSR) and/or with Ro318220(a specific inhibitor of PKC pathway).To evaluate the status of cardiac hypertrophy,cell diameter was observed by HE dyeing,and protein content was determined through coomassie brilliant blue protein kit.The intracellular calcium concentration( [ Ca2+]i) was determined by laser scanning confocal microscope.The protein expression of CaSR and PKC pathway were analyzed using Western blotting.Results ①Compared to the control group(0.1263 ± 0.0443),the protein expression of CaSR was increased in Ang Ⅱ group and in GdCl3 group(0.1963 ± 0.0375,0.2778 ± 0.0564,all P＜ 0.05).Moreover,compared with Ang Ⅱ alone,the increase was significant in GdCl3 group(P ＜ 0.05).②Compared to control group(222.70 ± 22.09),AngⅡ group(392.16 ± 36.85) remarkably increased [Ca2+]i(P＜ 0.05),and this increase of [Ca2+]i was further enhanced in GdCl3 group (502.60 ± 44.21) versus Ang Ⅱ group (P ＜ 0.05).③Compared to control group,Ang Ⅱ could induce cardiomyocyte hypertrophy,and GdCl3 enhanced the effect.Moreover,this enhancement was attenuated by Ro318220.④Compared to control group(0.27 ± 0.07,0.69 ± 0.06,0.87 ± 0.04),the protein expression of PKCα,PKCe and PKCδ was increased in Ang Ⅱ group(0.60 ± 0.16,1.02 ± 0.13,1.20 ± 0.18,all P＜ 0.05) and the protein expression of PKCα,PKCe was increased in GdCl3 group(0.82 ± 0.16,1.34 ± 0.12,all P ＜ 0.05).Moreover,compared with Ang Ⅱ group,the protein expression of PKCα,PKCe was obviously increased in GdCl3 group (all P ＜ 0.05);compared with GdCl3 group,the protein expression of PKCα,PKCe(0.41 ± 0.10,0.85 ± 0.14) was obviously decreased in Ro318220 group(all P ＜ 0.05).Conclusions CaSR is involved in cardiac hypertrophy induced by Ang Ⅱ through PKC pathway in cultured neonatal rat cardiomyocytes. Key words: Receptors, calcium-sensing; Protein kinase C; Angiotensin Ⅱ ; Cardiac hypertrophy

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钙敏感受体在新生大鼠血管紧张素II诱导心肌肥大中的作用

目的探讨钙敏感受体(CaSR)在血管紧张素Ⅱ(AngⅡ)心肌细胞肥厚模型中的作用及其可能机制。方法用AngⅡ处理培养的新生大鼠心室肌细胞作为细胞心肌肥大模型。用GdCl3(CaSR的特异性激动剂)和/或Ro318220(PKC通路的特异性抑制剂)处理肥厚新生大鼠心肌细胞。采用HE染色法观察细胞直径，考马斯亮蓝蛋白试剂盒测定蛋白质含量，评价心肌肥厚状态。用激光扫描共聚焦显微镜测定细胞内钙浓度([Ca2+]i)。Western blotting分析CaSR和PKC通路的蛋白表达。结果①与对照组(0.1263±0.0443)相比，AngⅡ组和GdCl3组CaSR蛋白表达量分别升高(0.1963±0.0375、0.2778±0.0564,P均< 0.05)。GdCl3组与单独使用AngⅡ组比较，差异有统计学意义(P < 0.05)。②与对照组(222.70±22.09)相比，AngⅡ组(392.16±36.85)明显升高[Ca2+]i(P< 0.05)， GdCl3组(502.60±44.21)较AngⅡ组进一步升高[Ca2+]i(P< 0.05)。③与对照组相比，AngⅡ能诱导心肌细胞肥大，GdCl3能增强作用。此外，这种增强被Ro318220减弱。④与对照组(0.27±0.07、0.69±0.06、0.87±0.04)相比，AngⅡ组PKCα、PKCe、PKCδ蛋白表达量升高(0.60±0.16、1.02±0.13、1.20±0.18，均P< 0.05)， GdCl3组PKCα、PKCe蛋白表达量升高(0.82±0.16、1.34±0.12，均P< 0.05)。与AngⅡ组相比，GdCl3组PKCα、PKCe蛋白表达量明显升高(P < 0.05);与GdCl3组相比，Ro318220组PKCα、PKCe蛋白表达量(0.41±0.10,0.85±0.14)明显降低(P < 0.05)。结论CaSR通过PKC途径参与AngⅡ诱导的新生大鼠心肌细胞心肌肥厚。关键词:受体;钙感应;蛋白激酶C;血管紧张素Ⅱ;心脏肥大

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来源期刊

中华地方病学杂志我国对人类健康危害特别严重的地方性疾病：克山病、大骨节病、碘缺乏病、地方性氟中毒、地方性砷中毒、鼠疫、布鲁氏菌病、寄生虫、新冠肺炎等疾病，同时还报道多发性自然疫源性疾病。

CiteScore

1.60

自引率

0.00%

发文量

8714

期刊介绍： The Chinese Journal of Endemiology covers predominantly endemic diseases threatening health of the people in the areas affected by the diseases including Keshan disease, Kaschin-Beck Disease, iodine deficiency disorders, endemic fluorosis, endemic arsenism, plague, epidemic hemorrhagic fever, brucellosis, parasite diseases and the diseases related to local natural and socioeconomic conditions; and reports researches in the basic science, etiology, epidemiology, clinical practice, control as well as multidisciplinary studies on the diseases.