长链非编码RNA巨噬细胞刺激1假基因2对缺氧诱导心肌细胞损伤的影响及机制  

Effects of long non-coding RNA macrophage stimulating 1 pseudogene 2 on hypoxia-induced myocardial cell injury and relevant mechanisms

作　　者：何忠开 郑重洲 安佰超 黎明 冯景焜 姚峰 HE Zhongkai;ZHENG Chongzhou;AN Baichao;LI Ming;FENG Jingkun;YAO Feng(Department of Structural Heart Disease,Cardiovascular Medicine Center,Affiliated Hospital of Guangdong Medical University,Zhanjiang 524001,Guangdong Province,China;School of Pharmacy,Guangdong Medical University,Dongguan 523808,Guangdong Province,China;School of Traditional Chinese Medicine,Southern Medical University,Guangzhou 510515,Guangdong Province,China;Cardiovascular Center of The Sixth Affiliated Hospital,South China University of Technology,Foshan 528200,Guangdong Province,China)

机构地区：[1]广东医科大学附属医院心血管中心结构性心脏病专科,广东湛江524001 [2]广东医科大学药学院,广东东莞523808 [3]南方医科大学中医药学院,广东广州510515 [4]华南理工大学附属第六医院心血管中心,广东佛山528200

出　　处：《新乡医学院学报》2025年第2期85-92,共8页Journal of Xinxiang Medical University

基　　金：广东省自然科学基金项目(编号:2018A030313355);湛江市科技计划项目(编号:2021B01073)。

摘　　要：目的探讨长链非编码RNA巨噬细胞刺激1假基因2(lncRNA MST1P2)对心肌细胞缺氧性损伤的影响及机制。方法将人心肌细胞株AC16细胞接种于不含胎牛血清的无糖达尔伯克改良伊格尔培养基(DMEM),置于37℃、含体积分数5%CO_(2)和1%O_(2)的培养箱培养0~48 h;分别于培养0、3、6、12、24、48 h采用实时荧光定量聚合酶链反应检测AC16细胞中lncRNA MST1P2及微RNA-133b(miR-133b)表达水平。将对数生长期AC16细胞随机分为正常对照组、缺氧组、缺氧+过表达对照组、缺氧+MST1P2过表达组,其中正常对照组、缺氧组细胞不做转染处理,缺氧+过表达对照组细胞转染空载体质粒,缺氧+MST1P2过表达组细胞转染lncRNA MST1P2过表达质粒,正常对照组细胞用低糖DMEM培养基在37℃、体积分数5%CO_(2)条件下培养,缺氧组、缺氧+过表达对照组、缺氧+MST1P2过表达组细胞用无糖DMEM培养基在37℃、含体积分数1%O_(2)和5%CO_(2)条件下缺氧处理24 h。应用细胞计数试剂盒-8检测4组细胞活性,流式细胞术检测4组细胞凋亡情况,Western blot法检测4组细胞中沉默信息调节因子-1(Sirt1)、磷酸化蛋白激酶B(p-Akt)及裂解的caspase-3(cleaved caspase-3)蛋白表达。结果缺氧处理0、3、6 h时AC16细胞中lncRNA MST1P2相对表达量比较差异无统计学意义(P>0.05);缺氧12、24、48 h时AC16细胞中lncRNA MST1P2的相对表达量显著低于缺氧处理0、3、6 h时(P<0.05);缺氧12 h与24 h时AC16细胞中lncRNA MST1P2的相对表达量比较差异无统计学意义(P>0.05);缺氧48 h时AC16细胞中lncRNA MST1P2的相对表达量显著低于缺氧12、24 h时(P<0.05)。缺氧处理0、3、6 h时AC16细胞中miR-133b相对表达量比较差异无统计学意义(P>0.05);缺氧12、24、48 h时AC16细胞中miR-133b的相对表达量显著高于缺氧处理0、3、6 h时(P<0.05);缺氧24 h与48 h时AC16细胞中miR-133b的相对表达量比较差异无统计学意义(P>0.05);缺氧24、48 h时AC16细胞中miR-133b�Objective To investigate the effects of long non-coding RNA macrophage stimulating 1 pseudogene 2(lncRNA MST1P2)on hypoxia-induced myocardial cell injury and relevant mechanisms.Methods AC16 cells were inoculated into the sugar-free Dulbecco′s modified Eagle′s medium(DMEM)without fetal bovine serum and then incubated in a 37℃incubator containing 5%CO_(2)and 1%O_(2) for 0-48 hours.The expression level of lncRNA MST1P2 and microRNA(miR)-133b in AC16 cardiomyocytes was detected by real-time quantitative polymerase chain reaction(RT-qPCR)after incubation for 0,3,6,12,24,and 48 hours,respectively.AC16 cardiomyocytes in the logarithmic growth phase were randomly divided into the normal control(NC)group,the hypoxia group,the hypoxia+overexpression control group,and the hypoxia+MST1P2 overexpression group.The cells in the NC group and the hypoxia group were not transfected,those in the hypoxia+overexpression control group were transfected with empty vector plasmids,and those in the hypoxia+MST1P2 overexpression group were transfected with lncRNA MST1P2 overexpression plasmids.The cells in the NC group were cultured in the low sugar DMEM under the condition of 37℃and 5%CO_(2)by volume,while those in the hypoxia group,the hypoxia+overexpression control group,and the hypoxia+MST1P2 overexpression group were cultured in the sugar-free DMEM medium under the condition of 37℃and 1%O_(2) and 5%CO_(2)by volume for 24 hours.The activity of cells in the four groups was detected by cell counting kit-8(CCK-8),the apoptosis of cells in the four groups was detected by flow cytometry(FC),and the expression of silent information regulator 1(Sirt1),phospho-Akt(p-Akt),and cleaved caspase-3 protein in the cells of the four groups was detected by Western blot(WB).Results There was no statistically significant difference in the relative expression level of lncRNA MST1P2 in AC16 cardiomyocytes treated with hypoxia for 0,3,and 6 hours(P>0.05).The relative expression level of lncRNA MST1P2 in AC16 cardiomyocytes treated with hypoxia fo

关 键 词：长链非编码RNA巨噬细胞刺激1假基因2 微RNA-133b 缺氧 凋亡 沉默信息调节因子-1/磷酸化蛋白激酶B信号通路 

分 类 号：R541.4[医药卫生—心血管疾病]