摘要
目的 探讨以腺病毒为载体的HIF-1α三突变体(Ad-HIF-1α-Trip)对高糖、低氧环境下H9c2心肌细胞凋亡的影响。方法 将体外培养的H9c2心肌细胞置于低氧浓度培养箱中培养,并随机分组,依据正常或高葡萄糖浓度的培养环境以及病毒转染情况分为4组,分别为低氧+正常葡萄糖浓度组(LO+NG组),低氧+高葡萄糖浓度组(LO+HG组),低氧+高糖+腺病毒空载体组(LO+HG+Ad-Null组),低氧+高糖+腺病毒HIF-1α三突变组(LO+HG+Ad-HIF-1α-Trip组)。低氧下培养12 h后,采用qRT-PCR、Western blot检测各组H9c2心肌细胞HIF-1α、PI3K和Akt在mRNA和蛋白水平的表达情况;采用流式细胞仪检测各组H9c2心肌细胞的凋亡情况。结果 qRT-PCR、Western blot检测显示高糖、低氧环境下HIF-1α、PI3K、Akt在心肌细胞中的表达下降,转染Ad-HIF-1α-Trip可增强其表达。流式细胞术分析显示,在高糖、低氧环境下,心肌细胞凋亡率增加,而LO+HG+Ad-HIF-1α-Trip组H9c2心肌细胞的凋亡率降低。结论 Ad-HIF-1α-Trip可增强高糖、低氧环境下H9c2心肌细胞HIF-1α、PI3K、Akt的表达,可能通过激活PI3K-Akt信号通路减少心肌细胞凋亡。
Objective To investigate the effect of the mutant of adenovirus-mediated HIF-1αtriple mutant(Ad-HIF-1α-Trip)on the apoptosis of H9c2 cardiomyocytes under high glucose and hypoxia conditions.Methods H9c2 cardiomyocytes were cultured under hypoxic conditions in vitro and randomly divided into four groups based on glucose concentration and viral transfection status:low oxygen+normal glucose concentration group(LO+NG group),low oxygen+high glucose concentration group(LO+HG group),low oxygen+high glucose+adenoviral null vector group(LO+HG+Ad-Null group),and low oxygen+high glucose+adenovirus HIF-1αtriple mutant group(LO+HG+Ad-HIF-1α-Trip group).After 12 hours of hypoxia,HIF-1α,PI3K,and Akt expression were measured via qRT-PCR and Western blot,and apoptosis of H9c2 cardiomyocytes was assessed by flow cytometry.Results HIF-1α,PI3K,and Akt expression decreased in cardiomyocytes under high glucose and hypoxia,while Ad-HIF-1α-Trip enhanced their expression.Flow cytometry revealed increased apoptosis under high glucose and hypoxia,which was reduced by Ad-HIF-1α-Trip.Conclusion Ad-HIF-1α-Trip upregulates HIF-1α,PI3K,and Akt in H9c2 cardiomyocytes under high glucose and hypoxia,likely reducing apoptosis via PI3K-Akt pathway activation.
作者
麦莉
潘永婷
何文凯
吴晓倩
李明琰
MAI Li;PAN Yongting;HE Wenkai;WU Xiaoqian;LI Mingyan(Department of Cardiology,the Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 511400,Guangdong,China;不详)
出处
《实用医学杂志》
北大核心
2025年第17期2617-2623,共7页
The Journal of Practical Medicine
基金
国家自然科学基金项目(编号:82370391)
广州市校(院)联合资助项目(编号:202201020135)。
