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加味丹参饮含药血清预处理对缺氧/复氧H9C2心肌细胞自噬相关基因Atg5和Beclin1 mRNA表达的影响

ISSN号：1005-9202
期刊名称：《中国老年学杂志》
时间：0
分类：R285.5[医药卫生—中药学;医药卫生—中医学]
作者机构：湖南中医药大学,湖南长沙410208
相关基金：国家自然科学基金资助项目（81373576,81503536）;湖南省中医药管理局重点课题（201304）;湖南省教育厅项目（14B136,14C0872）;湖南省教育厅优秀青年项目（14B136）;湖南省科技厅一般项目（2014SK3034）;中西医结合防治心脑疾病的相关基础研究湖南省高校科技创新团队科研项目

关键词：加味丹参饮含药血清, H9C2心肌细胞, 缺氧/复氧损伤, 自噬, Atg5, mRNA, BECLIN1, mRNA, Jiawei Danshen decoction medicated serum, H9C2 cardiomyocytes, Hypoxia/reoxygenation injury, Autophagy, Atg5 mRNA, Beclin1 mRNA

中文摘要：

目的探讨加味丹参饮含药血清预处理对缺氧/复氧H9C2心肌细胞自噬及其相关基因Atg5、Beclin1 mRNA表达的影响。方法将体外培养的H9C2心肌细胞随机分为空白血清组（CG）、缺氧/复氧组（HRG）、加味丹参饮含药血清组（JDG）、JDG＋自噬抑制剂（3-MA）组（JIG）、JDG＋自噬激动剂（RAPA）组（JAG）。倒置显微镜观察各组心肌细胞生长及形态变化;MTT比色法测定心肌细胞存活率;透射电子显微镜观察心肌细胞自噬体形态及数量;实时荧光定量PCR检测自噬相关基因Atg5、Beclin1 mRNA表达。结果与CG比较,HRG镜下心肌细胞变性坏死程度增加,细胞存活率下降（P〈0.01）,电镜下自噬体增多,实时荧光定量PCR示Atg5、Beclin1 mRNA表达上调;与HRG相比,JDG及JAG镜下心肌细胞变性坏死程度减轻,细胞存活率显著升高（P〈0.01）,电镜下自噬体增加,Atg5、Beclin1 mRNA表达进一步上调（P〈0.05）。结论加味丹参饮预处理通过上调自噬相关基因Atg5、Beclin1 mRNA表达,增强细胞自噬,保护缺氧/复氧损伤H9C2心肌细胞。

英文摘要：

Objective To investigate the influence of preconditioning with Jiawei Danshen decoction on expression level of autophagy related gene Atg5 and Beclin1 in H9C2 cardiomyocytes during hypoxia/reoxygenation injury.Methods H9C2 cardiomyocytes cultured in vitro were randomly divided into control group（ CG）,hypoxia/reoxygenation group（ HRG）,Jiawei Danshen decoction group（ JDG）,JDG ＋ 3-methyladenine（ a inhibitor of autophagy） group（ JIG） and JDG ＋Rapamycin（ RAPA）（ an agonists of autophagy） group（ JAG）. The inverted microscope was used to observe cell growth and morphological changes.The cell viability of H9C2 cardiomyocytes were measured by MTT assay.Transmission electron microscope was applied to observe the changes of autophagosome formation in cardiomyocytes.The expression levels of Atg5 and Beclin1 mRNA were determined by qRT-PCR.Results Compared with those of CG,the degree of cell necrosis and damage were rised in HRG,the cell viability was declined（ P〈0.01）,the autophagosome formation in cardiomyocytes was increased.Meanwhile,the expression levels of Atg5 and Beclin1 gene were up-regulated compared with those of HRG,the cardiomyocyte morphological structure of JDG and JAG were improved,the cell viability of JDG and JAG were increased significantly（ P〈0.01）,the autophagosome formation in cardiomyocytes were increased.In addition,the expression levels of Atg5 and Beclin1 gene were up-regulated（ P 〈 0. 05）. Conclusions Preconditioning with Jiawei Danshen decoction has a protective effect on H9C2 cardiomyocytes during hypoxia/reoxygenation injury by up-regulating the expression levels of Atg5 and Beclin1 gene.