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加味丹参饮预处理调控自噬相关基因Beclin-1和Atg5表达抗大鼠心肌缺血再灌注损伤

ISSN号：1674-070X
期刊名称：《湖南中医药大学学报》
时间：0
分类：R285[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
相关基金：国家自然科学基金资助项目(81373576;81503536);湖南省教育厅项目(14B136;14C0872);湖南省中医药管理局重点课题(201304);湖南省科技厅一般项目(2014SK3034);湖南省教育厅优秀青年项目(14B136);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, 心肌缺血再灌注, 自噬, BECLIN-1, Atg5, 丹参, 檀香, 川芎

中文摘要：

目的探讨加味丹参饮预处理是否通过调节Beclin-1和Atg5表达调控自噬抗缺血再灌注损伤大鼠心肌。方法将60只健康SD大鼠随机分为空白对照(control group,C)组、假手术(sham,S)组、缺血再灌注损伤(ischemia reperfusion injury,IRI)组、IRI+加味丹参饮(Jiawei Danshen Yin,JDY)组、IRI+JDY+自噬抑制剂(inhibitor,I)组,每组12只。通过结扎-放松大鼠左冠状动脉前降支制备心肌IRI模型。通过氯化三苯基四氮唑(TTC)染色观察心肌梗死面积率;在透射电镜下观察自噬泡;采用实时荧光定量逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)方法检测心肌Beclin-1和Atg5 m RNA表达;采用蛋白质印迹(western blot)法检测心肌Beclin-1蛋白表达变化。结果 IRI+JDY组心肌梗死面积率显著低于IRI组及IRI+JDY+I组(P<0.01)。电镜结果显示,IRI+JDY组适度调节大鼠缺血再灌注损伤心肌细胞的自噬,改善心肌细胞结构。IRI组大鼠心肌细胞中Beclin-1和Atg5 m RNA表达水平及Beclin-1蛋白表达较SG组显著升高(P<0.01);IRI+JDY组、IRI+JDY+I组大鼠心肌细胞中Beclin-1和Atg5 m RNA表达水平及Beclin-1蛋白表达较IRI组显著降低(P<0.01);IRI+JDY组大鼠心肌细胞中Beclin-1和Atg5 m RNA表达水平及Beclin-1蛋白表达较IRI+JDY+I组显著升高(P<0.01)。结论加味丹参饮通过调节缺血再灌注损伤大鼠心肌细胞自噬相关基因Beclin-1和Atg5表达适度,调控缺血再灌注心肌细胞发生适度自噬,从而发挥细胞保护作用。

英文摘要：

〔Abstract〕 Objective To investigate the pretreatment with Jiawei Danshen decoction against myocardial ischemia reperfusion injury in rats whether by regulating Beclin-1 and Atg5 expression of autophagy. Methods The 60 healthy SD rats were randomly divided into control (C) group, sham-operation (S) group, ischemia reperfusion injury (IRI) group, IRI+Jiawei Danshen decoction (JDS) group, IRI+JDY+Inhibitor (I) group, 12 rats in each group. The IRI model was established by ligating-relaxing the left anterior descending coronary of rats. Myocardial infarction rate was measured by using triphenyltetrazolium chloride (TTC) staining. The autophagic vacuoles were observed via transmission electron microscope (TEM). The mRNA expression of autophagy related gene Beclin-1 and Atg5 were detected by real-time quantitative reverse transcription- polymerase chain reaction (RT-PCR), and the expression changes of myocardial Beclin-1 protein were detected by Western blot. Results The infarction rate inIRI+JDY group was significantly smaller than that in IRI group and IRI+JDY+I group (P<0.01). TEM showed that in IRI+JDY group, autophagy of cardiomyocytes could be regulated moderately and the structure of cardiomyocytes could be improved. The expression levels of Beclin-1 and Atg5 mRNA, and Beclin-1 protein in myocardium of rat in IRI group were significantly higher than SG group (P<0.01). The expression levels of Beclin-1 and Atg5 mRNA, and the expression of Beclin-1 protein in myocardium of rats in IRI+JDY group and IRI+JDY+I group were decreased significantly compared with the IRI group (P<0.01). The expression levels of Beclin-1 and Atg5 mRNA, and Beclin-1 protein in myocardium of rats with IRI+JDY group were significantly higher than that in IRI+JDY+I group (P<0.01). Conclusion The modified Danshen decoction can regulate the autophagy of ischemia reperfusion cardiomyocytes and protect the cells by regulating the autophagy related genes (Beclin-1 and Atg5).

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