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

关键词:  蕨麻  内皮细胞  缺氧  缺氧诱导因子-1α  内皮素-1

Key words:  Potentilla anserina L.  endothelial cell  hypoxia  hypoxia inducible factor-1  endothelin-1

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)mRNA和内皮素-1(ET-1)mRNA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着降低,HIF-1α 和ET-1 mRNA 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,细胞存活率显着升高,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 mRNA及蛋白水平显着降低(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。

Abstract:

[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-1α and ET-1 mRNA were detected by reverse transcription-PCR assay. The Western blot and imuunocytochemistry were used for determining HIF-1α protein. The ET-1 protein was detected 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 products 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-1α pathway.