miR-483-3p对乏氧/复氧诱导的心肌细胞凋亡和焦亡的作用研究  

Effects of miR⁃483⁃3p on hypoxia/reoxygenation⁃induced apoptosis and pyroptosis in cardiomyocytes

作　　者：卢钰芬 郑孝明 韦少娟[1] 陈礼琴 徐彤彤[1] 吕祥威 LU Yufen;ZHENG Xiaoming;WEI Shaojuan;CHEN Liqin;XU Tongtong;LÜXiangwei(Department of General Medicine,Affiliated Hospital of Guilin Medical College,Guilin 541001,Guangxi,China;不详)

机构地区：[1]桂林医学院附属医院综合科,广西桂林541001 [2]桂林医学院广西肿瘤免疫与微环境调控重点实验室,广西桂林541199

出　　处：《实用医学杂志》2025年第3期339-346,共8页The Journal of Practical Medicine

基　　金：国家自然科学基金项目(编号:82160068,82260864)。

摘　　要：目的探讨miR-483-3p对乏氧/复氧(hypoxia/reoxygenation,H/R)诱导的H9c2心肌细胞凋亡和焦亡的影响及其可能机制。方法体外培养大鼠H9c2心肌细胞,腺相关病毒感染H9c2及三气培养箱构建H/R模型,将细胞随机分为空白对照(Sham)组、模型(H/R)组、AAV-miR-483-3p mimic+H/R(AAV-miR-483-3p)组、AAV-miR-483-3p阴性对照+H/R(AAV-NC)组。使用倒置显微镜观察各组细胞生长状态;细胞计数试剂盒-8(CCK-8)法检测细胞增殖活性;乳酸脱氢酶(LDH)试剂盒检测各组细胞LDH释放量;流式细胞术检测各组细胞凋亡率;缺口末端标记法(TUNEL)法检测各组细胞凋亡情况;蛋白免疫印迹法(Western blot)检测各组细胞IL-1β、GSDMD蛋白的表达水平。结果与Sham组相比,H/R组细胞状态异常且细胞死亡数量增多、细胞活性下降、LDH释放量增多、细胞凋亡率及凋亡水平升高、IL-1β、GSDMD蛋白的表达水平升高(P<0.05);与H/R组相比较,AAV-miR-483-3p组细胞状态得到改善且细胞死亡数量较少、细胞增殖活性上升、LDH释放量下降、细胞凋亡率及凋亡水平降低、IL-1β及GSDMD蛋白的表达水平降低(P<0.05)。结论过表达miR-483-3p可通过提升细胞活性、细胞代谢、抑制细胞凋亡及细胞焦亡的机制改善H/R诱导的H9c2心肌细胞损伤。Objective To investigate the effects of miR-483-3p on hypoxia/reoxygenation(H/R)-induced apoptosis and pyroptosis of H9c2 cardiomyocytes and its possible mechanism.Methods Rat H9c2 cardiomyocytes were cultured in vitro,adeno-associated virus-infected H9c2 and the H/R model were constructed by triple-air incubator,and the cells were randomly divided into blank control(Sham)group,model(H/R)group,AAV-miR-483-3p mimic+H/R(AAV-miR-483-3p)group,AAV-miR-483-3p negative control+H/R(AAV-NC)group.The growth status of cells in each group was observed using an inverted microscope;cell proliferation activity was detected by cell counting kit-8(CCK-8);LDH release by lactate dehydrogenase(LDH)kit;apoptosis rate by flow cytometry;apoptosis by notched end labeling(TUNEL).Western blot(WB)was used to detect the expression levels of IL-1βand GSDMD proteins in each group.Results Compared with the Sham group,the H/R group showed abnormal cell status and increased cell death,decreased cell activity,increased LDH release,increased apoptosis rate and apoptosis level,and increased expression levels of IL-1βand GSDMD proteins(P<0.05);compared with the H/R group,the AAV-miR-483-3p group showed improved cell status and less cell death,increased cell proliferation activity,increased LDH release,and increased IL-1βand GSDMD protein expression levels(P<0.05).Compared with the H/R group,the AAV-miR-483-3p group showed improved cell status and less cell death,increased cell proliferation activity,decreased LDH release,decreased apoptosis rate and apoptosis level,and decreased expression of IL-1βand GSDMD proteins(P<0.05).Conclusion Over-expression of miR-483-3p can improve H/R-inducedH9c2 cardiomyocyte injury by enhancing cell activity and cell metabolism,and inhibiting apoptosis and cell charring.

关 键 词：miR-483-3p 心肌缺血再灌注损伤 细胞凋亡 细胞焦亡 

分 类 号：R318.08[医药卫生—生物医学工程]