超极化激活环核苷酸门控阳离子通道4在大鼠心肌成纤维细胞和心肌细胞中的表达  

Expression of hyperpolarization-activated cyclic nucleotide gated cation channel 4 in cardiac fibroblasts and cardiomyocytes of rats

作　　者：贾丹丹 许亚平 郭志坤[1,2] Jia Dandan;Xu Yaping;Guo Zhikun(Henan Key Laboratory of Medical Tissue Regeneration,Xinxiang Medical University,Xinxiang 453003;Zhengzhou Seventh People's Hospital,Zhengzhou 450000,China)

机构地区：[1]新乡医学院河南医用组织再生重点实验室,新乡453003 [2]郑州第七人民医院,郑州450000

出　　处：《解剖学杂志》2023年第2期105-109,F0002,F0003,共7页Chinese Journal of Anatomy

基　　金：国家自然科学基金(81570268)。

摘　　要：目的:探讨超极化激活环核苷酸门控阳离子通道4(HCN4)在大鼠体内和体外心肌成纤维细胞、心肌细胞中的表达分布及其增龄变化。方法:SD大鼠分为新生组(1~3 d),成年组(17个月)和老年组(22个月),每组8只,其中4只用于冰冻切片,另4只用于PCR;取新生SD大鼠心,酶消化法分离培养原代心肌成纤维细胞和原代心肌细胞。用免疫荧光、免疫组织化学检测原代心肌成纤维细胞和原代心肌细胞HCN4的表达;免疫印迹、实时荧光定量PCR检测原代心肌成纤维细胞HCN4的表达,用实时荧光定量PCR检测不同月龄大鼠心肌组织HCN4的表达。结果:体外细胞培养显示,成纤维细胞和原代心肌细胞中均表达HCN4。组织切片显示,新生、成年和老年大鼠心肌成纤维细胞和部分心肌细胞表达HCN4。P1~P4代成纤维细胞的免疫印迹和实时荧光定量PCR结果显示HCN4随着成纤维细胞代次增加,表达量逐渐降低;不同月龄大鼠左心室的实时荧光定量PCR结果显示随着大鼠月龄增加,HCN4的表达量逐渐降低,其结果与培养的细胞表达一致。结论:HCN4在心肌成纤维细胞和部分心肌细胞中表达。随增龄变化,心肌细胞和成纤维细胞HCN4的表达量逐渐下降。Objective:To detect the expression and aging of hyperpolarization-activated cyclic nucleotide gated cation channel 4(HCN4)in myocardial fibroblasts and cardiomyocytes in vitro and in vivo.Methods:SD rats were divided into the newborn group(1-3 days),the adult group(17 months),and the elderly group(22 months),with eight rats in each group.Four of them were used for frozen sections and the other four were used for real-time fluorescence quantitative polymerase chain method(qRT-PCR).Cardiac fibroblasts and cardiomyocytes were isolated and cultured in 15 newborn SD rats by enzyme digestion.The expression of HCN4 in primary cardiac fibroblasts and primary cardiomyocytes was detected by immunofluorescence and immunohistochemistry.The expression of HCN4 in primary cardiac fibroblasts was detected by Western blotting and qRT-PCR,and the expression of HCN4 in myocardium of SD rats of different age was detected by qRT-PCR.Results:HCN4 was expressed in fibroblasts and primary cardiac myocytes in vitro.Tissue sections showed that HCN4 was expressed in cardiac fibroblasts and some cardiac myocytes of newborn,adult and aged rats.Western blotting and qRT-PCR results of P1-P4 generation fibroblasts showed that the expression level of HCN4 gradually decreased with the increase of fibroblast generation,and the qRT-PCR results of left ventricle of rats at different months of age showed that the expression level of HCN4 gradually decreased with the increase of month of age,which was consistent with the expression level of cultured cells.Conclusion:HCN4 is expressed in fibroblasts and some cardiac myocytes,and the expression of HCN4 in cardiac myocytes and fibroblasts decreases gradually with age.

关 键 词：超极化激活环核苷酸门控阳离子通道4 成纤维细胞 心肌细胞 心 大鼠 

分 类 号：R542.2[医药卫生—心血管疾病]