葱白提取物调控HIF-1α/VEGF通路抑制缺氧/复氧心肌细胞的氧化应激和凋亡  

Fistular onion bulb regulated HIF-1α/VEGF pathway for arresting oxidative stress and apoptosis of hypoxic/reoxygenated cardiomyocytes

作　　者：吴勇宏 贺立群[1] 潘婉[1] 田立群[1] 卜文玉 康棋棋 刘翰 WU Yonghong;HE Liqun;PAN Wan;TIAN Liqun;BU Wenyu;KANG Qiqi;LIU Han(Department of Cardiovascular Medicine,First Municipal Hospital,Hubei Wuhan 430022,China;College Ⅰ of Clinical Medicine,Hubei University of Chinese Medicine,Hubei Wuhan 430070,China)

机构地区：[1]武汉市第一医院心血管内科,湖北武汉430022 [2]湖北中医药大学第一临床学院,湖北武汉430070

出　　处：《中国医院药学杂志》2024年第23期2725-2730,共6页Chinese Journal of Hospital Pharmacy

基　　金：武汉市卫生健康委科研项目面上项目(编号:WZ21C39)。

摘　　要：目的:评估葱白提取物(fistular onion bulb,FOB)通过调控HIF-1α/VEGF通路对缺氧/复氧H9c2心肌细胞氧化应激和凋亡的影响。方法:基于H9c2大鼠心肌细胞构建H/R细胞模型,通过细胞形态观察和CCK-8法检测确定细胞复氧时间为24 h。将细胞分为Control组、H/R组、H/R+FOB组、H/R+YC-1(HIF-1α抑制剂)组、H/R+YC-1+FOB组。CCK-8法检测各组细胞增殖活力,流式细胞术检测细胞凋亡及细胞中活性氧(ROS)含量,生化试剂盒检测细胞中丙二醛(MDA)、乳酸脱氢酶(LDH)含量,Western blot检测细胞中Bax、Bcl-2、低氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)和血管内皮生长因子(vascular endothelial growth factor,VEGF)的蛋白表达水平。结果:与Control组比较,H/R组细胞增殖活力下降(P<0.05),凋亡率上升(P<0.05),ROS、MDA和LDH含量上升(P<0.05),凋亡相关蛋白Bax表达上升(P<0.05)、Bcl-2表达下降(P<0.05),HIF-1α/VEGF通路蛋白表达均上升(P<0.05)。与H/R组比较,H/R+FOB组和H/R+YC-1组细胞增殖活力增加(P<0.05),凋亡率下降(P<0.05),ROS、MDA和LDH含量下降(P<0.05),Bcl-2蛋白表达增加(P<0.05),Bax、HIF-1α、VEGF蛋白表达均下降(P<0.05)。与H/R+YC-1组比较,H/R+YC-1+FOB组变化趋势更显著。结论:FOB通过抑制HIF-1α/VEGF通路,从而抑制缺氧/复氧心肌细胞的氧化应激和凋亡,减轻细胞损伤。OBJECTIVE To evaluate the effects of fistular onion bulb(FOB)extract on oxidative stress and apoptosis in hypoxia/reoxygenated H9c2 cardiomyocytes through regulating HIF-1α/VEGF pathway.METHODS Hypoxia/reoxygenation(H/R)cell model was constructed by H9c2 rat cardiomyocytes with a reoxygenation time of 24 h as determined by cell morphology observation and CCK-8 assay.The cells were assigned into five groups of control,H/R,H/R+FOB,H/R+YC-1(HIF-1αinhibitor)and H/R+YC-1+FOB.Cell proliferation activity was assessed by CCK-8 while cell apoptosis by flow cytometry.The contents of reactive oxygen species(ROS)were detected by flow cytometry and malondialdehyde(MDA)and level of lactate dehydrogenase(LDH)by biochemical kits.Western blot was utilized for determining the protein expression levels of Bax,Bcl-2,hypoxia-inducible factor-1α(HIF-1α)and vascular endothelial growth factor(VEGF).RESULTS As compared with control group,cell proliferation activity and apoptotic rate decreased in H/R group(P<0.05)while the contents of ROS,MDA and LDH rose significantly(P<0.05).Additionally,apoptosis-related protein Bax was up-regulated(P<0.05)while Bcl-2 down-regulated(P<0.05).Protein expression levels of HIF-1α/VEGF pathway became elevated(P<0.05).As compared with H/R group,cell proliferation activity spiked obviously in both H/R+FOB and H/R+YC-1 groups(P<0.05)while apoptotic rate decreased(P<0.05).Furthermore,the contents of ROS,MDA and LDH all declined(P<0.05)while Bcl-2 protein was up-regulated(P<0.05).The protein expressions of Bax,HIF-1αand VEGF all decreased(P<0.05).CONCLUSION FOB may suppress the activation of HIF-1α/VEGF pathway,resulting in lower oxidative stress and slower apoptosis within hypoxic/reoxygenated cardiomyocytes,ultimately alleviating cellular injury.

关 键 词：心肌梗死 葱白提取物 缺氧/复氧 氧化应激 细胞凋亡 HIF-1α/VEGF通路 

分 类 号：R285.5[医药卫生—中药学] R963[医药卫生—中医学]