甲基化转移酶样蛋白3在急性心肌梗死患者外周血中的表达及对缺氧缺血所致心肌细胞损伤的调节作用  

Expression of peripheral blood methyltransferase-like protein 3 in patients with acute myocardial infarction and its regulatory effect on hypoxia/ischemia-induced myocardial damage

作　　者：周钰[1] 白峰华[2] 张君[3] 詹峰[3] 董小莉[4] 吴奎 ZHOU Yu;BAI Feng-hua;ZHANG Jun;ZHAN Feng;DONG Xiao-li;WU Kui(Admission Service Center,Hainan Provincial People′s Hospital,Haikou 570311,China;Department of Science and Education,Hainan Provincial People′s Hospital,Haikou 570311,China;Emergency ICU Ward,Hainan Provincial People′s Hospital,Haikou 570311,China;Department of Cardiology,Hainan Provincial People′s Hospital,Haikou 570311,China;School of Basic Medicine and Life Science,Hainan Medical College,Haikou 571199,China)

机构地区：[1]海南省人民医院入院服务中心,海口市570311 [2]海南省人民医院科教处,海口市570311 [3]海南省人民医院急诊ICU病房,海口市570311 [4]海南省人民医院心血管内科,海口市570311 [5]海南医学院基础医学与生命科学学院,海口市571199

出　　处：《广西医学》2021年第9期1095-1100,1148,共7页Guangxi Medical Journal

基　　金：海南省卫生健康行业科研项目(19A200042)。

摘　　要：目的探讨甲基化转移酶样蛋白3(METTL3)在急性心肌梗死患者外周血中的表达及对急性缺氧/缺血(HI)所致心肌细胞损伤的调节作用。方法(1)收集60例急性心肌梗死患者和60例健康者的血浆和血清,采用酶联免疫吸附测定法和实时荧光定量聚合酶链反应法分别检测血清和血浆中METTL3的表达水平。(2)取人心肌细胞AC16,分为对照组、HI-2h组、HI-4h组、siMETTL3+HI-2h组、siNC+HI-2h组;对照组不进行任何处理,HI-2h组、HI-4h组使用1%O 2和1%胎牛血清培养基诱导HI心肌细胞模型,siMETTL3+HI-2h组、siNC+HI-2h组分别转染有效沉默METTL3的小干扰RNA序列、阴性对照小干扰RNA序列后建立HI心肌细胞模型。检测各组AC16细胞的增殖率和凋亡率,METTL3、激活型天冬氨酸特异性半胱氨酸蛋白酶(cleaved-Caspase3)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)。将溶酶体抑制剂巴弗洛霉素A1添加至培养基后,按照上述方法进行分组及干预,检测微管相关蛋白1轻链3B-Ⅱ(LC3B-Ⅱ)、人死骨片1(SQSTM1)的蛋白表达水平。结果(1)心肌梗死患者血浆METTL3 mRNA相对表达水平以及血清中METTL3表达水平均高于健康者(P<0.05)。(2)与对照组比较,HI-2h组、HI-4h组AC16细胞的增殖率降低,细胞凋亡率升高,METTL3蛋白、cleaved-Caspase3蛋白、Bax/Bcl-2比值、SQSTM1蛋白的表达上调,而LC3B-Ⅱ的蛋白表达下调(均P<0.05)。与HI-2h组和siNC+HI-2h组比较,siMETTL3+HI-2h组的细胞增殖率升高,细胞凋亡率降低,METTL3、cleaved-Caspase3、Bax/Bcl-2比值、SQSTM1的蛋白表达下调,LC3B-Ⅱ的蛋白表达上调(均P<0.05)。结论METTL3在心肌梗死患者体内和HI诱导的急性心肌细胞损伤模型中的表达均升高,沉默METTL3可以抑制HI心肌细胞的凋亡并增加自噬通量,从而促进细胞增殖的恢复。Objective To explore the expression of peripheral blood methyltransferase-like protein 3(METTL3)in patients with acute myocardial infarction and its regulatory effect on acute hypoxia/ischemia(HI)-induced myocardial damage.Methods(1)Plasma and serum were collected from 60 patients with acute myocardial infarction and 60 healthy individuals,enzyme-linked immunosorbent assay and real-time quantitative PCRwere separately used to detect the expression levels of serum and plasma METTL3.(2)Human myocardiocytes AC16 were collected and divided into control group,HI-2h group,HI-4h group,siMETTL3+HI-2h group,and siNC+HI-2h group.The control group did not receive any treatment,the HI-2h and HI-4h groups used culture medium containing 1%O 2 and 1%fetal calf serum to induce a myocardiocyte model of HI,the siMETTL3+HI-2h group and the siNC+HI-2h group were modeled as myocardiocytes of HI afteRtransfection with small interfering RNA sequence effectively silencing METTL3 and negative control small interfering RNA sequence,respectively.The proliferation rate and apoptosis rate of AC16 cells,METTL3,cleaved cysteinyl aspartate specific proteinase(cleaved-Caspase),B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)were detected in groups.AfteRlysosome inhibitoR bafilomycin A1 was added to culture medium,grouping and intervention were performed according to the procedures mentioned above,the expression levels of microtubule-associated protein 1 light chain 3B-Ⅱ(LC3B-Ⅱ)and human sequestosome 1(SQSTM1)proteins were detected.Results(1)Patients with myocardial infarction had higheR relative expression level of plasma METTL3 mRNA and serum METTL3 expression level than healthy individuals(P<0.05).(2)Compared with the control group,the HI-2h and HI-4h groups exhibited loweRproliferation rate of AC16 cells,increased apoptosis rate,up-regulated expression of METTL3,cleaved-Caspase3 and SQSTM1 proteins,higheRBax/Bcl-2 ratio,and down-regulated LC3B-Ⅱprotein expression(all P<0.05).The siMETTL3+HI2h group had increased proliferatio

关 键 词：急性心肌梗死 急性缺血缺氧损伤 N 6-甲基腺苷 甲基化转移酶样蛋白3 细胞增殖 细胞凋亡 细胞自噬 

分 类 号：R542.22[医药卫生—心血管疾病]