亚精胺通过miR-451介导自噬缓解阿霉素诱导的心肌细胞衰老  

Spermidine Mediates Autophagy Through miR-451 to Alleviate Adriamycin-induced Cardiomyocyte Senescence

作　　者：戴婷 徐亚宁 谌赟 蔡毅 刘敏 Dai Ting;Xu Yaning;Chen Yun(General Department,Wuhan Sixth Hospital,Affiliated Hospital of Jianghan University,Wuhan 43014,China)

机构地区：[1]江汉大学附属医院,武汉市第六医院综合科,武汉430014

出　　处：《华中科技大学学报(医学版)》2024年第6期791-795,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：武汉市科创局项目(No.2020020601012316);武汉市卫健委医学科研项目(No.WG20M02)。

摘　　要：目的评估亚精胺(spermidine,SPD)对miR-451-5p的调控作用及通过调控miR-451-5p对衰老的心肌细胞增殖活性和自噬的影响。方法阿霉素处理大鼠心肌H9C2细胞,构建心肌衰老细胞模型。将细胞分为对照(Control)组,模型(Model)组,阴性对照(NC)+SPD组和miR-451-5p抑制剂(Inhibitor)+SPD组。除Control组外,其余组均采用阿霉素构建心肌衰老细胞模型,NC+SPD组和Inhibitor+SPD组细胞造模后分别转染miR-NC和miR-451-5p inhibitor,并用10μmol/L的SPD处理24 h。CCK-8检测细胞增殖活性,β-半乳糖苷酶染色(SA-β-gal)检测心肌细胞衰老情况,Western blot检测细胞中心肌衰老标志物P21蛋白的表达,qRT-PCR检测细胞中miR-451-5p、自噬标志物(LC3Ⅱ和Beclin1)的mRNA表达。结果与正常H9C2细胞相比,心肌衰老细胞模型中miR-451-5p低表达(P<0.05)。与Control组相比,Model组细胞增殖活性下降(P<0.05),SA-β-gal阳性的染色面积增加,P21蛋白表达上调(均P<0.05),miR-451-5p、LC3Ⅱ和Beclin1 mRNA表达下调(均P<0.05)。与Model组相比,NC+SPD组细胞增殖活性增加,SA-β-gal阳性的染色面积减少(均P<0.05),P21蛋白表达下调(P<0.05),miR-451-5p、LC3Ⅱ和Beclin1 mRNA表达上调(均P<0.05)。与NC+SPD组相比,Inhibitor+SPD组细胞增殖活性下降(P<0.05),SA-β-gal阳性的染色面积增加,P21蛋白表达上调(均P<0.05),miR-451-5p、LC3Ⅱ和Beclin1 mRNA表达下调(均P<0.05)。结论亚精胺可通过上调miR-451-5p的表达促进心肌衰老模型细胞的增殖和自噬,进而缓解心肌细胞的衰老。Objective To study the regulatory effect of spermidine(SPD)on miR-451-5p and the effect of autophagy mediated by miR-451-5p on proliferation and apoptosis of aging cardiomyocytes.Methods Doxorubicin was used to treat H9C2 cells in rat myocardium,and cardiac senescence cell model was constructed.The cells were divided into Control group,Model group,negative control(NC)+SPD group and miR-451-5p Inhibitor+SPD group.Cardiac senescent cell models were constructed with adriamycin in all groups except Control group.After modeling,cells of NC+SPD group and Inhibitor+SPD group were transfected with miR-NC and miR-451-5p inhibitor,respectively,and treated with 10μmol/L SPD for 24 h.CCK-8 was used to detect cell proliferative activity;β-galactosidase staining was used to detect myocardial senescence;Western blot was used to detect the protein expression of P21,a marker of aging in heart tissue;and qRT-PCR was used to detect mRNA expression of miR-451-5p and autophagy markers(LC3Ⅱand Beclin1).Results Compared with normal H9C2 cells,the expression of miR-451-5p was lower in cardiac senescent cell models(P<0.05).Compared with Control group,cell proliferation activity in Model group was decreased(P<0.05),SA-β-gal positive staining area increased,and P21 protein expression was up-regulated(P<0.05).The miR-451-5p expression,mRNA expressions of LC3Ⅱand Beclin1 were down-regulated(P<0.05).Compared with model group,cell proliferation activity in NC+SPD group was increased(P<0.05),SA-β-gal positive staining area decreased,and P21 protein expression was down-regulated(P<0.05).The miR-451-5p expression,mRNA expressions of LC3Ⅱand Beclin1 were up-regulated(P<0.05).Compared with NC+SPD group,cell proliferation activity of Inhibitor+SPD group was decreased(P<0.05).SA-β-gal positive staining area increased,and P21 protein expression was up-regulated(P<0.05).The miR-451-5p expression,mRNA expressions of LC3Ⅱand Beclin1 were down-regulated(P<0.05).Conclusion Spermidine can promote the proliferation and autophagy of cardiac senes

关 键 词：心肌细胞 亚精胺 miR-451 自噬 细胞衰老 

分 类 号：R542.2[医药卫生—心血管疾病]