基于Nrf2-ARE信号通路探讨维立西呱减轻心肌细胞缺氧复氧损伤作用机制OA
Mechanism of vericiguat Alleviating Hypoxic Reoxygenation Injury of Cardiomyocytes Based on Nrf2-ARE Signaling Pathway
目的:探究基于 Nrf2-ARE 信号通路探讨维立西呱减轻心肌细胞缺氧复氧损伤作用机制.方法:常规培养 H9C2 细胞,分对照组(常规培养)、缺氧-复氧组(无血清 DMEM 培养液+94%N2+5%CO2+1%O2 及 95%空气培养)、维立西呱低、中剂量组、高剂量组(分别添加 1μmoL/L、3μmoL/L、10μmoL/L 维立西呱培养).TBA 法测定 H9C2 细胞中 SOD、MDA 活性,流式细胞仪检测细胞 ROS 含量,克隆形成实验检测 H9C2 细胞克隆情况,划痕实验检测 H9C2 细胞迁移功能,流式细胞仪检测 H9C2细胞凋亡,Western blot 法检测 H9C2 细胞中 Nrf2、ARE 蛋白.结果:与对照组比较,缺氧-复氧组 MDA、ROS、细胞凋亡率升高,GSH、细胞克隆率、Nrf2、ARE 降低(P<0.05),与缺氧-复氧组比较,维立西呱-低剂量组 MDA、ROS、细胞凋亡率降低,GSH、细胞克隆率、Nrf2、ARE 升高(P<0.05);与维立西呱-低剂量组比较,维立西呱-中剂量组 MDA、ROS、细胞凋亡率降低,GSH、细胞克隆率、Nrf2、ARE 升高(P<0.05);与维立西呱-中剂量组比较,维立西呱-高剂量组 MDA、ROS、细胞凋亡率降低,GSH、细胞克隆率、Nrf2、ARE 升高(P<0.05).划痕实验结果显示,在0h 各组空白区域无显著差异;24h 后,缺氧-复氧组空白区增大,维立西呱低、中、高剂量组空白区域缩小,其中维立西呱-高剂量组空白区域总体变窄显著,提示降低缺氧/复氧导致的细胞迁移功能缺陷显著.结论:维立西呱对于改善心肌细胞缺氧复氧损伤具有一定积极效用,并可能通过调节 Nrf2-ARE 信号通路发挥干预效用,这一发现可为临床治疗心肌缺血再灌注损伤提供新的思路和潜在治疗靶点.
Objective:To explore the mechanism of vericiguat alleviating hypoxic reoxygenation injury of cardiomyocytes based on Nrf2-ARE signaling pathway.Methods:H9C2 cells were routinely cultured and di-vided into control group(routine culture),hypoxic-reoxygenation group(no serum DMEM culture medium+94%N2+5%CO2+1%O2 and 95%air culture),low dose group,medium dose group and high dose group(adding 1μmol/L,3μmol/L and 10μmol/L vericiguat culture,respectively).SOD and MDA activities in H9C2 cells were determined by TBA method,ROS content was detected by flow cytometry,colony formation assay was used to detect H9C2 cell colony formation,scratch assay was used to detect H9C2 cell migration function,and flow cytometry was used to detect H9C2 cell apoptosis.Nrf2 and ARE proteins in H9C2 cells were detected by Western blot.Results:Compared with control group,MDA,ROS and apoptosis rates were increased in hypoxia-reoxygenation group,while GSH,cell colony formation rate,Nrf2 and ARE were de-creased(P<0.05).Compared with hypoxia-reoxygenation group,MDA,ROS and apoptosis rates were de-creased in low dose vericiguat group,while GSH,cell colony formation rate,Nrf2 and ARE were increased(P<0.05);Compared with low dose vericiguat group,MDA,ROS and apoptosis rate were decreased in me-dium dose vericiguat group,while GSH,cell colony formation rate,Nrf2 and ARE were increased(P<0.05).Compared with medium dose vericiguat group,MDA,ROS and apoptosis rates were decreased in high dose vericiguat group,while GSH,cell colony formation rate,Nrf2 and ARE were increased(P<0.05).The scratch test results showed that there was no significant difference in the blank area among all groups at 0h.After 24h,the blank area increased in the hypoxic-reoxygenation group,and decreased in the low,medium and high dose groups of vericiguat,among which the blank area narrowed significantly in the high dose ver-iciguat group,suggesting that vericiguat significatly alleviated hypoxia/reoxygenation-induced cell migration dysfunction.Conclusion:Vericeiquat has positive effect on alleviating hypoxic-reoxygenation injury in cardio-myocytes,and may exert intervention effect by regulating Nrf2-ARE signaling pathway.This finding can pro-vide new ideas and potential therapeutic targets for clinical treatment of myocardial ischemia-reperfusion inju-ry.
张珍;李翩;刘凯;张欢;方媛
华中科技大学同济医学院附属梨园医院,湖北 武汉 430077华中科技大学同济医学院附属梨园医院,湖北 武汉 430077华中科技大学同济医学院附属梨园医院,湖北 武汉 430077华中科技大学同济医学院附属梨园医院,湖北 武汉 430077华中科技大学同济医学院附属梨园医院,湖北 武汉 430077
心肌细胞维立西呱Nrf2ARE损伤
VericiguatCardiomyocytesvericiguatNrf2AREInjury
《河北医学》 2026 (6)
949-954,6
国家自然科学基金项目(82004309)
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