心肌纤维化模型大鼠心肌组织中GRK2的表达以及对胶原合成的影响  

Effect and expression of GRK2 on collagen synthesis in myocardial fibrosis rat model

作　　者：阚丽丽[1] 蒋方国[1] 安丰双[2] 

机构地区：[1]兖矿集团有限公司总医院重症医学科,山东邹城273500 [2]山东大学齐鲁医院心内科,济南250012

出　　处：《山东大学学报（医学版）》2013年第9期22-25,30,共5页Journal of Shandong University:Health Sciences

摘　　要：目的探讨G蛋白偶联受体激酶2(GRK2)在心肌纤维化发生过程中的作用。方法皮下注射异丙肾上腺素诱导大鼠心肌纤维化,采用HE染色观察模型大鼠心肌组织学变化,差速贴壁法分离培养心肌成纤维细胞,检测成纤维细胞中GRK2的表达和胶原Iα含量,采用siRNA干涉法抑制GRK2表达,采用cAMP检测试剂盒检测细胞内cAMP含量。结果 HE染色显示大鼠心肌纤维化模型心肌细胞出现坏死、肥大、淋巴细胞浸润等变化。Western blotting结果显示,异丙肾上腺素诱导模型大鼠心肌成纤维细胞中GRK2表达增加(P<0.05),胶原Iα表达增加(P<0.01)。GRK2特异性siRNA可以显著抑制GRK2的表达(P<0.05)。cAMP含量分析显示,异丙肾上腺素诱导心肌纤维化模型大鼠中心肌组织cAMP含量明显下降(P<0.01),在siRNA抑制GRK2表达后,心肌组织中cAMP水平显著升高(P<0.05)。结论采用siRNA法抑制GRK2表达可以增加细胞内cAMP的含量。GRK2有望成为心肌纤维化防治靶点。Objective To study the role of G protein-coupled receptor kinase 2 （GRK2） on myocardial fibrosis. Meth- ods Rat model of myocardial fibrosis was established by subcutaneous injection with isoproterenol. Myocardial histo- logical changes of the rats were tested by microscope after hematoxylin and eosin staining （HE）. Myocardial fibroblasts were isolated and cultured with differential anchoring velocity. The expressions of GRK2 and collagen type Is were examined by western blotting. The cAMP level was tested after GRK2 gene was silenced by siRNA. Results HE stai- ning showed myocardium necrosis, hypertrophy, and lymphocytic infiltration. The expression of GRK2 （ P 〈 0.05 ） and collagen type IoL （ P 〈 0.05 ） were up-regulated in isoproterenol-induced rats. The level of cAMP decreased significantly in isoproterenol-induced rats （ P 〈 0.05 ）. However, the level of cAMP increased after GRK2 gene silenced by siRNA （ P 〈 0.05 ）. Conclusion The intracellular cAMP increased after GRK2 gene expression is silenced by siRNA. GRK2 may serve as a novel potential therapeutic target.

关 键 词：心肌纤维化 G蛋白偶联受体激酶2 环磷酸腺苷 

分 类 号：R542.2[医药卫生—心血管疾病]