冠心舒通胶囊调节PI3K/AKT/mTOR信号通路保护H9c2心肌细胞机制研究  

Guanxin Shutong capsule treats damage protection mechanism of H9c2 cardiomyocytes through PI3K/AKT/mTOR pathway

作  者:后亚芳 刘园 陆晨曦 段婷婷 任菲菲 杜霞[2] HOU Yafang;LIU Yuan;LU Chenxi;DUAN Tingting;REN Feifei;DU Xia(Shaanxi University of Chinese Medicine,Xianyang 712046,China;Shaanxi Academy of Traditional Chinese Medicine,Xi’an 710003,China)

机构地区:[1]陕西中医药大学,陕西咸阳712046 [2]陕西省中医药研究院,陕西西安710003

出  处:《陕西中医》2025年第3期296-300,共5页Shaanxi Journal of Traditional Chinese Medicine

基  金:国家自然科学基金青年科学基金资助项目(82204711);陕西省中医药管理局“双链融合”中青年科研创新团队(2022-SLRH-YQ-003);陕西省中医药管理局“医研校企”中医药传承创新平台(中医药创新药物/器械“研发-转化-推广”平台)项目。

摘  要:目的:本研究探讨冠心舒通胶囊基于PI3K/AKT/mTOR通路防治H9c2心肌细胞缺氧/复氧损伤(H/R)的作用机制。方法:建立H9c2心肌细胞H/R模型,分为对照组、模型组、冠心舒通胶囊不同浓度组。采用PI染色法(PI)和酶标法测定乳酸脱氢酶(LDH)活性、肌酸激酶同工酶(CK-MB)含量评价模型。冠心舒通胶囊组采用梯度浓度对细胞进行预给药处理,采用CCK-8法测定细胞活性,酶标法测定LDH活性、CK-MB含量,并通过实时荧光定量PCR(RT-qPCR)法检测磷酸肌醇3-激酶(PI3K)、蛋白激酶B(AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)的mRNA水平。结果:CCK-8法筛选出缺氧8、16 h细胞用于后续实验检测。PI染色4、8μg/ml冠心舒通胶囊组细胞活性在60%~75%;与模型组相比,经冠心舒通胶囊干预后H/R H9c2心肌细胞活性显著提升,而LDH活性、CK-MB含量降低,PI3K、AKT、mTOR的mRNA表达降低。结论:冠心舒通胶囊通过调节PI3K/AKT/mTOR信号通路抑制H9c2细胞损伤,对H/R造成的心肌损伤具有一定保护作用。Objective:To investigate mechanism of Guanxin Shutong capsule in prevention and treatment of hypoxia reoxygenation cardiomyocytes(Hypoxia/Reoxygenation,H/R)based on PI3K/AKT/mTOR pathway.Methods:H9c2 cardiomyocyte H/R model was established and randomly divided into control group,model group and Guanxin Shutong capsule group with different concentration.The model was evaluated by PI staining method,enzyme labelling to determine lactate dehydrogenase(LDH)activity and creatine kinase isoenzymes(CK-MB)content.Guanxin Shutong capsule group were treated with gradient concentration of pre-administration of cells,cell viability determined by cell counting kit-8(CCK-8),LDH activity and CK-MB content determined by enzymatic hydrogenase method.The mRNA levels of Phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT)and mechanistic target of rapamycin(mTOR)were detected by RT-qPCR.Results:Cells with 8 h and 16 h of hypoxia were screened by CCK-8 method for follow-up test.The cell viability of the 4μg/ml and 8μg/ml Guanxin Shutong capsule groups stained by PI was 60%~75%.Compared with the model group,after intervention of Guanxin Shutong capsule,the viability of H9c2 cardiomyocytes were significantly improved,LDH activity and CK-MB content reduced,and mRNA expressions of PI3K,AKT and mTOR significantly decreased.Conclusion:Guanxin Shutong capsule can inhibit injury of H9c2 cells by regulating PI3K/AKT/mTOR signaling pathway,and has certain protective effect on myocardial injury caused by hypoxia/reoxygenation.

关 键 词:冠心病 冠心舒通胶囊 H9C2心肌细胞 H/R模型 PI3K/AKT/mTOR信号通路 心肌损伤 

分 类 号:R256[医药卫生—中医内科学]

 

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