中药药理与临床2025，Vol.41Issue(11)：33-38,6.
康达心调控AMPK/mTOR介导自噬减轻异丙肾上腺素诱导心肌细胞损伤
Regulatory Effect of Kangdaxin on Myocardial Injury Induced by Isoprenaline Based on AMPK/mTOR-Mediated Autophagy
摘要
Abstract
Objective:To investigate the mechanism of Kangdaxin(康达心)on alleviation of myocardial injury induced by isoprenaline(ISO)based on the autophagy mediated by adenosine 5'-monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR).Methods:The Kangdaxin-containing serum was prepared,and the optimum concentration and time of treatment were screened.H9c2 cardiomyocytes were pretreated with different concentrations of Kangdaxin-containing serum for two hours and then treated with 10 μmol/L ISO for 24 hours.The cell counting kit-8(CCK8)was used to determine the cell survival rate,and the flow cytometry was em-ployed to detect the cell apoptosis.The polymerase chain reaction(PCR)method was used to detect the expression of atrial natriuretic pep-tide(Anp),and western blot was used to detect the expressions of p-AMPK/AMPK,p-mTOR/mTOR,microtubule associated protein 1 light chain 3(LC3)-Ⅱ,LC3-Ⅰ,and autophagy regulatory protein-1(BECLIN-1).Transmission electron microscopy was used to observe the autoph-agosomes.The AMPK inhibitor Dorsomorphin was used to further verify the relationship between Kangdaxin and AMPK/mTOR signaling pathway.Results:Compared with that in the complete medium group,the cell survival rate showed no obvious change in the 10%and 5%18 g/kg Kangdaxin-containing serum groups.The 10%Kangdaxin-containing serum intervention for 24 h was selected as the optimal experi-ment condition,while 5%Kangdaxin-containing serum was selected as the gradient comparison concentration.Compared with the blank con-trol group,the model control group exhibited significantly reduced cell survival rate,increased cell apoptosis rate and Anp gene expression,down-regulated p-AMPK protein expression,up-regulated p-mTOR protein,decreased autophagy-related protein expressions(LC3 Ⅱ/Ⅰ and BE-CLIN-1),and reduced number of autophagosomes(P<0.05 or P<0.01),which suggested a successful myocardial injury model.Compared with the model control group,the 5%and 10%18 g/kg Kangdaxin-containing serum groups showed significantly increased cell survival rate,decreased cell apoptosis rate and Anp gene expression,significantly elevated p-AMPK protein and autophagy-related protein(LC3 Ⅱ/Ⅰ and BE-CLIN-1)expressions,significantly increased number of autophagosomes,and down-regulated p-mTOR protein expression(P<0.05).After being treated with the AMPK inhibitor Dorsomorphin,the effect of Kangdaxin in activating autophagy and reducing myocardial injury was weakened(P<0.05).Conclusion:Kangdaxin can induce autophagy and attenuate ISO-induced myocardial injury.Its mechanism may be as-sociated with the AMPK/mTOR signaling pathway.关键词
康达心/自噬/腺苷酸活化蛋白激酶/哺乳动物雷帕霉素靶蛋白Key words
Kangdaxin(康达心)/Autophagy/Adenosine 5'-monophosphate-activated protein kinase/Mammalian target of rapamycin引用本文复制引用
林飞宁,黄丽华,林超,杨直,李翠云,熊尚全..康达心调控AMPK/mTOR介导自噬减轻异丙肾上腺素诱导心肌细胞损伤[J].中药药理与临床,2025,41(11):33-38,6.基金项目
福建省自然科学基金项目(编号:2023J01830) （编号:2023J01830）
福建中医药大学优势学科专项课题(编号:X2022008-专项). （编号:X2022008-专项）
