亚精胺通过PI3K/AKT/mTOR通路调节自噬抑制心肌细胞肥大的机制研究  

Mechanism of autophagy inhibition of cardiomyocyte hypertrophy mediated by spermidine through PI3K/AKT/mTOR signaling pathway

作　　者：朱磊[1] 谌赟 童婷 谈文娟 程波[1] 刘敏[1] ZHU Lei;CHEN Yun;TONG Ting;TAN Wenjuan;CHENG Bo;LIU Min(Department of General Practice,Affiliated Hospital of Jianghan University,Hubei Wuhan 430015,China)

机构地区：[1]江汉大学附属医院全科医学科,湖北武汉430015

出　　处：《中国医院药学杂志》2024年第22期2563-2568,2575,共7页Chinese Journal of Hospital Pharmacy

基　　金：2020年武汉市科创局项目(编号:2020020601012316);武汉市卫健委科研项目(编号:WX19C10)。

摘　　要：目的:探讨亚精胺通过调节自噬对心肌细胞肥大的作用及其机制。方法:异丙肾上腺素(isoprenaline,ISO)处理心肌细胞H9c2,构建心肌细胞肥大模型。RT-qPCR检测心肌肥大标志基因心房钠尿肽(atrial natriuretic peptide,ANP)和β型肌球蛋白重链(β-myosin heavy chain,β-MHC)mRNA的表达,进行模型鉴定。不同浓度亚精胺处理H9c2细胞,测量各组细胞表面积,CCK-8检测细胞活力,RT-qPCR检测细胞中ANP和β-MHC mRNA的表达,Western blot检测细胞中自噬相关蛋白LC3-Ⅰ、LC3-Ⅱ的水平。为进一步验证亚精胺对PI3K/AKT/mTOR信号通路的影响,用PI3K激活剂740Y-P处理细胞,流式细胞术检测细胞凋亡,RT-qPCR检测细胞中ANP和β-MHC mRNA的表达,Western blot检测细胞中自噬相关蛋白LC3-Ⅰ、LC3-Ⅱ、p62和PI3K/Akt/mTOR信号通路相关蛋白PI3K、p-PI3K、mTOR、p-mTOR的表达。结果:亚精胺干预后能够提高细胞活力和细胞内LC3-Ⅱ/Ⅰ水平(P<0.01),减少细胞表面积,降低ANP和β-MHC mRNA的表达(P<0.05),且呈剂量依赖性。PI3K激活剂740Y-P干预后,细胞凋亡率、细胞中ANP和β-MHC mRNA表达水平、p62和p-mTOR蛋白水平升高(P<0.01),LC3-Ⅱ/Ⅰ水平降低(P<0.01),而SPD与740Y-P联合干预能够逆转740Y-P的上述作用(P<0.01)。结论:亚精胺能够通过激活细胞自噬抑制心肌细胞肥大,其机制可能与抑制PI3K/Akt/mTOR信号通路有关。OBJECTIVE To explore the effect of spermidine(SPD)on cardiomyocyte hypertrophy through regulating autophagy and elucidate its mechanism.METHODS Cardiomyocyte H9c2 was treated with isoprenaline(ISO)and a hypertro-phic model of cardiomyocyte established.The mRNA expressions of atrial natriuretic peptide(ANP)and beta-myosin heavy chain(β-MHC)markers of myocardial hypertrophy were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and the model was verified.At different concentrations of SPD,cell surface area of each group was measured.Cellular viability was examined by CCK-8,the expressions of ANP andβ-MHC mRNA were measured by RT-qPCR and the expressions of autophagy related proteins LC3-Ⅰand LC3-Ⅱwas detected by Western blot.To further verify the effect of SPD on PI3K/AKT/mTOR signaling pathway,cells were treated with PI3K activator 740Y-P.Cellular apoptosis was examined by flow cytom-etry,the expressions of ANP andβ-MHC mRNA were quantified by RT-qPCR and the expressions of autophagy related proteins LC3-Ⅰ,LC3-Ⅱ,p62 and PI3K/Akt/mTOR signaling pathway related proteins PI3K,p-PI3K,mTOR and p-mTOR were detected by Western blot.RESULTS Spermidine intervention could improve cellular viability and intracellular LC3-Ⅱ/Ⅰlevels(P<0.01),reduce cell surface area and down-regulate the expressions of ANP andβ-MHC mRNA(P<0.05)in a dose-dependent manner.After 740Y-P intervention,apoptotic rate,mRNA expression levels of ANP/β-MHC and protein expression levels of p62 and p-mTOR rose(P<0.01)while LC3-Ⅱ/Ⅰlevel declined(P<0.01).The combined intervention of SPD and 740Y-P could reverse the above effects of 740Y-P(P<0.01).CONCLUSION Spermidine can arrest myocardial hypertrophy through activating autophagy.Its mechanism may be related to an inhibition of PI3K/Akt/mTOR signaling pathway.

关 键 词：亚精胺 自噬 心肌肥大 PI3K/AKT/mTOR信号通路 

分 类 号：R96[医药卫生—药理学]