N6-甲基腺嘌呤-胰岛素样生长因子-2 mRNA结合蛋白1通过促进甾醇邻酰基转移酶1表达调节心力衰竭的机制研究  

N6-methyladenosine-Insulin-like growth factor-Ⅱ binding protein l exacerbates heart failure by promoting sterol O-acyltransferase 1 expression

作　　者：钟妮尔 王菲 邓纪钊 杨光[1] Zhong Nier;Wang Fei;Deng Jizhao;Yang Guang(Department of Cardiology,Shaanxi Provincial People's Hospital,Xi'an 710068,China)

机构地区：[1]陕西省人民医院心血管内二科,西安710068

出　　处：《临床内科杂志》2025年第3期237-242,共6页Journal of Clinical Internal Medicine

基　　金：陕西省自然科学基础研究计划项目(2024JC-YBMS-670);陕西省人民医院科技人才支持计划项目(2022BJ-16、2023JY-49)。

摘　　要：目的 探讨胰岛素样生长因子-2 mRNA结合蛋白1(IGF2BP1)和甾醇邻酰基转移酶1(SOAT1)在心力衰竭(HF)中的分子机制。方法 分离获取大鼠心肌细胞,将其分为对照组、缺氧模型组、sh-NC组、sh-SOAT1组、sh-METTL3组、sh-METTL3+Vector组、sh-METTL3+pcDNA-SOAT1组、sh-IGF2BP1组、sh-IGF2BP1+Vector组、sh-IGF2BP1+pcDNA-SOAT1组。对照组细胞在正常氧环境培养,其余各组均在缺氧环境培养。采用Me-RIP分析SOAT1 mRNA的N6-甲基腺嘌呤(m6A)富集水平;RIP分析验证SOAT1 mRNA与METTL3和IGF2BP1蛋白的结合情况;检测细胞活力、细胞凋亡率、活性氧(ROS)、乳酸脱氢酶(LDH)、葡萄糖摄取、IL-1β及肿瘤坏死因子(TNF)-α水平。通过横向主动脉收缩(TAC)手术构建HF模型,将64只雄性SD大鼠随机分为假手术组、HF组、HF+LV-NC组、HF+sh-SOAT1组、HF+sh-SOAT1+METTL3组和HF+sh-SOAT1+IGF2BP1组,术后均饲养4周。采用超声心动图分析左心室的功能,2,3,5-氯化三苯基四氮唑(TTC)染色分析梗死面积,Western blot检测SOAT1、METTL3和IGF2BP1蛋白表达水平。结果 与对照组比较,缺氧模型组SOAT1的mRNA和蛋白表达水平均显著升高,细胞活力和葡萄糖摄取水平减少,细胞凋亡率、ROS、LDH、IL-1β、TNF-α水平均升高;与sh-NC组比较,sh-METTL3组、sh-IGF2BP1组和shSOAT1组SOAT1的mRNA和蛋白表达水平均降低,细胞活力和葡萄糖摄取水平增多,细胞凋亡率、ROS、LDH、IL-1β、TNF-α水平均降低(P <0.05)。与sh-NC组比较,sh-METTL3组Anti-METTL3抗体下拉的复合物中m6A修饰的SOAT1 mRNA减少,sh-IGF2BP1组SOAT1 mRNA的半衰期更短;分别与sh-METTL3+vector组和sh-IGF2BP1+vector组比较,sh-METTL3+pcD NA-SOAT1组和shIGF2BP1+pcD NA-SOAT1组细胞活力均更低、细胞凋亡均更高(P <0.05)。动物实验结果显示,与假手术组比较,HF组大鼠心脏组织中SOAT1、METTL3和IGF2BP1蛋白表达水平均升高,心脏梗死面积增大,心脏功能发生障碍[左心室射血分数(LVEF)、左心室短轴�ObjectiveTo investigate the role of Insulin-like growth factor-II binding protein 1(ICF2BP1)and sterol O-acyltransferase 1(SOAT1)in heart failure(HF).Methods Rat cardiomyocytes were isolated and divided into control group,hypoxia model group,sh-NC group,sh-SOAT1 group,sh-METTL3 group,sh-METTL3+Vector group,sh-METTL3+pcDNA-SOAT1 group,sh-ICF2BP1 group,sh-IGF2BP1+Vector group,sh-IGF2BP1+pcDNA-SOAT1 group.The cells in control group were cultured in normal oxygen environment,and other groups were cultured in hypoxic environment.N6-methyladenosine(m6A)level of SOAT1 mRNA was analyzed by Me-RIP,and the bindings of METTL3 and IGF2BP1 to SOAT1 mRNA were verified by RIP.Cell viability,cell apoptosis,reactive oxygen species(ROS),lactate dehydrogenase(LDH),glucose uptake,IL-1β,and tumor necrosis factor(TNF)-αlevels were detected.HF model was constructed by transverse aortic constriction(TAC)surgery,and 64 male SD rats were randomly divided into sham group,HF group,HF+LV-NC group,HF+sh-SOAT1 group,HF+sh-SOAT1+METTL3 group and HF+sh-SOAT1+IGF2BP1 group,4 weeks of post operative feeding in each group.The function of left ventricle was analyzed by echocardiography;infarct volume was analyzed by 2,3,5-Triphenyl tetrazolium chloride(TTC)staining;protein levels of SOAT1,METTL3 and ICF2BP1 were detected by Western blot.Results Compared with control group,mRNA and protein levels of SOAT1 in hypoxia model group were significantly increased,cell viability and glucose uptake were decreased,and cell apoptosis rate,ROS,LDH,IL-1β,and TNF-αlevels were increased;compared with sh-NC group,sh-METTL3 group,sh-IGF2BP1 group,and the sh-SOAT1 group mRNA and protein levels of SOAT1 were decreased,cell viability and glucose uptake were increased,and cell apoptosis rate,ROS,LDH,IL-1β,and TNF-αlevels were decreased(P<0.05).Compared with sh-NC group,the m6A-modified SOAT1 mRNA in the complex of Anti-METTL3 in sh-METTL3 group was reduced,and the half-life of SOAT1 mRNA in sh-IGF2BP1 group was shorter.The cell viability of sh-METTL3+vector group a

关 键 词：心力衰竭 RNA N6腺苷甲基化 甾醇邻酰基转移酶1 甲基转移酶样蛋白3 胰岛素样生长因子-2 mRNA结合蛋白1 

分 类 号：R394.3[医药卫生—医学遗传学] R541[医药卫生—基础医学]