银杏二萜内酯对缺氧处理人脐静脉内皮细胞凋亡和血管生成的影响及机制  

Effect of ginkgo diterpene lactones on hypoxia-induced apoptosis and angiogenesis of human umbilical vein endothelial cells and its mechanisms

作　　者：石晓明[1] 杨永宾[1] 赵伟[1] 安彦波 吕柏楠[1] SHI Xiao-ming;YANG Yong-bin;ZHAO Wei;AN Yan-bo;LÜ Bo-nan(Department of Vascular Surgery,Hebei General Hospital,Shijiazhuang 050051,Hebei,China)

机构地区：[1]河北省人民医院血管外科,石家庄050051

出　　处：《第二军医大学学报》2020年第10期1123-1128,共6页Academic Journal of Second Military Medical University

基　　金：河北省中医药管理局科研计划项目(2016177)。

摘　　要：目的探讨银杏二萜内酯对缺氧处理的人脐静脉内皮细胞(HUVEC)凋亡和血管生成的影响及分子机制。方法在低氧状态下培养HUVEC 24 h,然后分别以低剂量(6.25 mg/L)、高剂量(25.00 mg/L)银杏二萜内酯进行处理。采用MTT法检测细胞活性,流式细胞术检测细胞凋亡情况,Transwell小室实验检测细胞迁移能力,qRT-PCR和蛋白质印迹法分别检测缺氧诱导因子1α(HIF-1α)、凋亡相关基因[Bcl-2、Bcl-2相关X蛋白(Bax)]及血管生成相关基因[血管内皮生长因子(VEGF)、TGF-β]的mRNA和蛋白表达。结果与正常氧培养组相比,低氧培养24 h后HUVEC的活性降低(P<0.05),细胞凋亡率增高(P<0.05),HIF-1α的mRNA和蛋白表达均增高(P均<0.05)。低剂量及高剂量银杏二萜内酯处理组HUVEC活性、细胞迁移能力均高于低氧处理组(P均<0.05),细胞凋亡率均低于低氧处理组(P均<0.05);同时高剂量银杏二萜内酯处理组HUVEC活性、细胞迁移能力均高于低剂量组(P均<0.05),细胞凋亡率低于低剂量组(P<0.05)。低、高剂量银杏二萜内酯处理组细胞HIF-1α、Bax的mRNA和蛋白表达均低于低氧处理组,Bcl-2、VEGF、TGF-β的mRNA和蛋白表达均高于低氧处理组,并且这些基因的表达变化在高剂量组更明显。结论银杏二萜内酯能通过调控HIF-1α及细胞凋亡、血管生成相关基因表达改善缺氧处理HUVEC的凋亡及血管生成能力,其可能成为缺氧性血管疾病的有效治疗药物。Objective To investigate the effect of ginkgo diterpene lactones on hypoxia-induced apoptosis and angiogenesis of human umbilical vein endothelial cells(HUVECs)and the related molecular mechanisms.Methods HUVECs were cultured under hypoxia for 24 h,and then treated with ginkgo diterpene lactones(low-dose:6.25 mg/L and high-dose:25.00 mg/L).MTT assay was used to detect the cell activity.Flow cytometry was used to detect the apoptosis of HUVECs.Transwell assay was employed to detect the migration of HUVECs.Quantitative real-time polymerase chain reaction and Western blotting were used to test the expression levels of mRNA and protein of hypoxia-inducible factor 1α(HIF-1α),apoptosis-related genes(B-cell lymphoma 2[Bcl-2]and B-cell lymphoma 2-related X protein[Bax]),and angiogenesisrelated genes(vascular endothelial growth factor[VEGF]and transforming growth factorβ[TGF-β]).Results Compared with the normal group,the HUVEC activity was significantly decreased after exposed to hypoxia for 24 h(P<0.05),and the apoptosis rate of HUVECs and mRNA and protein expression levels of HIF-1αwere significantly higher(all P<0.05).The cell activity and migration ability of HUVECs were significantly higher in the low-and high-dose ginkgo diterpene lactones groups than those in the hypoxia group(all P<0.05),and the apoptosis rates of HUVECs were significantly lower than those in the hypoxia group(both P<0.05).Meanwhile,the cell activity and migration ability were significantly higher in the highdose group than those in the low-dose group(both P<0.05),and the apoptosis rate was significantly lower than that in the low-dose group(P<0.05).The mRNA and protein expression levels of HIF-1αand Bax were lower in the low-and high dose ginkgo diterpene lactones groups than those in the hypoxia group,and the mRNA and protein expression levels of Bcl-2,VEGF and TGF-βwere higher than those in the hypoxia group;and the expression changes of the above genes were more significant in the high-dose group.Conclusion Ginkgo diterpene lactones can

关 键 词：银杏二萜内酯 低氧 人脐静脉内皮细胞 细胞凋亡 血管生成 

分 类 号：R543.6[医药卫生—心血管疾病]