蕨麻正丁醇部位下调缺氧内皮细胞HIF-1α及ET-1表达  

Study of N-butanol extract of Potentilla anserina L. in down-regulating expression levels of HIF-1α and ET-1 in hypoxia endothelial cells

作　　者：杨硕[1,2] 张岭[2] 龚海英[2] 张永亮 李灵芝[2,3] 高宏生 

机构地区：[1]天津医科大学,天津300070 [2]武警后勤学院药物化学教研室,天津300309 [3]天津市职业与环境危害防制重点实验室,天津300309

出　　处：《天津中医药》2015年第3期168-172,共5页Tianjin Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81073152);天津市重点基金项目(12JCZDJC34700);中国博士后基金项目(20100470106)

摘　　要：[目的]探讨蕨麻正丁醇部位对内皮细胞缺氧损伤的保护机制。[方法]采用人脐静脉内皮细胞(EA.hy926)建立缺氧损伤实验模型,实验设常氧对照组、缺氧模型组、高(3.00 g/L)、中(1.50 g/L)、低浓度(0.75 g/L)蕨麻正丁醇部位组,复方丹参(3.0 g/L)组。胎盘兰染色法测定各组细胞存活率;逆转录-聚合酶链反应(RT-PCR)技术检测各组缺氧诱导因子-1α(HIF-1α)m RNA和内皮素-1(ET-1)m RNA表达,免疫细胞化学染色及Western Blot方法检测HIF-1α蛋白表达。放免法测定培养基中ET-1的活性。[结果]与对照组比较,缺氧模型组细胞存活率显著降低,HIF-1α和ET-1m RNA及蛋白表达增加(P<0.01);蕨麻正丁醇部位各剂量组与缺氧模型组比较,细胞存活率显著升高,高、中、低剂量蕨麻正丁醇部位组、复方丹参组细胞HIF-1α和ET-1 m RNA及蛋白水平显著降低(P<0.01)。[结论]在缺氧状态下,蕨麻正丁醇部位可能通过HIF-1α途径调控靶基因的表达,从而发挥保护作用。Objective] To study the protective effect of n-butanol extract of Potentilla anserine L. on EA.hy926 endothelial cells exposed to hypoxia. [Methods] The model of hypoxia injury was established with human umbilical vein endothelial cell line （EA.hy926）. The cell death was determined by Trypan blue staining assay. The expression levels of HIF-1αand ET-1 mRNA were detected by reverse tran-scription-PCR assay. The Western blot and imuunocytochemistry were used for determining HIF-1αprotein. The ET-1 protein was de-tected with radioimmunoassay. [Results] Compared with control group, the survival rate of cell was significantly increased after pretreated with 3, 1.5 and 0.75 g/L of n-butanol extract of Potentilla anserine L. Meanwhile, the mRNAs of HIF-1 and ET-1 and their protein prod-ucts were significantly decreased. [Conclusion] Potentilla anserina L. could exert its protective effect on endothelial cells during hypoxia injury by modulating expression of target genes through HIF-1αpathway.

关 键 词：蕨麻 内皮细胞 缺氧 缺氧诱导因子-1α 内皮素-1 POTENTILLA anserina L. hypoxia INDUCIBLE factor-1 ENDOTHELIN-1 

分 类 号：R285.5[医药卫生—中药学]