文献类型：期刊文献

中文题名：当归补血汤超滤物抗H_2O_2致人脐静脉内皮细胞(ECV-304)氧化损伤的影响

英文题名：Effects of Ultra-filtration Extract from Danggui Buxue Tang against Human Umbilical Vein Endothelial Cells Injured by H_2O_2

作者：李应东[1,2];刘凯[1];赵信科[2]

第一作者：李应东

机构：[1]甘肃中医学院,甘肃兰州730000;[2]甘肃中医学院附属医院,甘肃兰州730000

第一机构：甘肃中医药大学

年份：2012

卷号：23

期号：1

起止页码：71

中文期刊名：时珍国医国药

外文期刊名：Lishizhen Medicine and Materia Medica Research

收录：北大核心:【北大核心2011】；CSCD:【CSCD_E2011_2012】；

基金：国家自然科学基金(No.81160478/H2902);甘肃省自然科学基金(No.1010RJZA174)

语种：中文

中文关键词：当归补血汤超滤物;人脐静脉内皮细胞;过氧化氢;血管紧张素Ⅱ受体1

外文关键词：Ultra-filtration extract from Danggui Buxue Tang Human umbilical vein endothelial cells Hydrogen dioxide（H2O2） Angiotensin Ⅱ receptor 1（AT1）

摘要：目的研究当归补血汤超滤物对H2O2诱导人脐静脉内皮细胞(human umbilieal vein endothelial cells,HUVECs)ECV-304氧化损伤的影响,并初步探讨其作用机制。方法通过酶消化法对离体的人脐静脉内皮细胞系ECV-304细胞进行消化传代,建立过氧化氢(peroxide hydrogen,H2O2)诱导的ECV-304细胞氧化损伤模型。采用超滤技术对当归红芪合剂进行分离与纯化,以不同分子量(2万以下、5万以下、10万以下)且浓度均为15.0 g/L的当归红芪超滤膜提取物作用于氧化损伤的ECV-304细胞,黄嘌呤氧化酶法检测细胞内超氧化物歧化酶(superoxide dismutase,SOD)活力,硫代巴比妥酸(TBA)法检测细胞内丙二醛(malondialdeyde,MDA)活力,流式细胞仪(flow cytometry,FCM)分析细胞周期的变化,RT-PCR检测细胞内AT1 mRNA的表达。结果经0.5mmol/L H2O2刺激后细胞活性明显降低,SOD活力明显降低,MDA活力明显提高,G0/G1期细胞的数目明显增加,S期细胞的数目明显减少,AT1 mRNA表达增加(P﹤0.05);与H2O2组相比,当归红芪超滤膜提取物可以增强H2O2诱导ECV-304细胞氧化损伤的细胞活性,增加细胞的存活率且以作用72 h为佳,提高氧化损伤的ECV-304细胞的SOD活力,降低氧化损伤的ECV-304细胞的MDA活力,促进氧化损伤的ECV-304细胞向S期转变,减少细胞内AT1 mRNA表达(P﹤0.01,P﹤0.05)。结论当归补血汤超滤物对氧化损伤ECV-304细胞的保护作用可能与其提高了抗氧化酶活性,降低了脂质过氧化物活性,促进了内皮细胞DNA合成复制及减少了细胞内AT1 mRNA的表达有关。
Objective To investigate the effect of ultra-filtration extract from the Danggui Buxue Tang（EDBT）on the oxidative stress of human umbilical vein endothelial cells（HUVECs） induced by H2O2,and to observe the mechanism of action of EAH. Methods HUVECs were digested by trypsin,and were transfering cultured in vitro,established to the model of the oxidative stress of HUVECs induced by H2O2.The EDBT was separated and purified by the ultra-filtration technique.The effect of EDBT on the oxidative stress of HUVECs induced by H2O2 was observed by following five means：the cell activity and survival ratio were measured by Methyl Thiazolyl Tetrazolium（MTT）,the activity of superoxide dismutase（SOD） were measured by xanthine oxidase,the activity of malondialdeyde（MDA） were measured by Thibabituric Acid（TBA）,using flow cytometry（FCM） to analyze the cell cycle and using RT-PCR to measure the expression of AT1 mRNA. Results H2O2（0.5mmol/L） could obiviously lower the cellular activity and the activity of SOD,raise the content of MDA,reduce the number of cells in S,increase the number of cells in G0/G1（P﹤0.01） and increase the expression of AT1 mRNA（P﹤0.05）;Compared with the model（H2O2） group,the EDBT could raise the cell activity,increase the survival ratio of cells with the better effect in 72h,raise the activity of SOD,lower the activity of MDA,increase the number of cells in S,reduce the number of cells in G0/G1 and reduce the expression of AT1 mRNA（P﹤0.05）. Conclusion The protective effects of EAH on HUVECs have business with the following factors：raising the activity of SOD,lowering the content of MDA,promoting DNA＇s synthesis and reproducation and reducing the expression of AT1 mRNA.