蓬子菜总黄酮对氧化损伤人脐静脉内皮细胞NF-κB/IκB信号途径的影响  

Effects of Total Flavones of Galium verum on NF-κB/IκB Signal Pathway in Human Umbilical Vein Endothelial Cells Injured by Hydrogen Dioxide

作　　者：张紫阳[1] 马英丽[2] 董晶[1] 李瑞[1] 董均明 朱玉良[1] 马玲[1] 

机构地区：[1]哈尔滨医科大学流行病教研室,哈尔滨150081 [2]黑龙江中医药大学药学院,哈尔滨150040

出　　处：《中国实验方剂学杂志》2015年第1期107-111,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81274034)

摘　　要：目的:研究蓬子菜总黄酮(FGV)对过氧化氢(H2O2)损伤的人脐静脉内皮细胞(HUVECs)保护作用的机制。方法:采用体外培养的HUVECs,取对数生长期的细胞调整细胞密度为7.5×104/m L,置培养箱中培养至细胞达80%融合时,将细胞随机分为5组:空白组、模型组(仅加入浓度为100μmol·L-1的H2O2培养4 h),FGV保护组(质量浓度分别为12.5,25,50 mg·L-1的FGV培养24 h,之后加入浓度为100μmol·L-1的H2O2培养4 h),每组设4个复孔。采用四甲基偶氮唑蓝(MTT)比色法检测细胞增殖抑制率;荧光分光光度法测定超氧化物歧化酶(SOD)的活性以及丙二醛(MDA)的含量;酶联免疫分析方法(ELISA)检测血栓调节蛋白(TM)和血管内皮细胞生长因子(VEGF)的分泌水平。逆转录聚合酶链式反应(RT-PCR)检测核因子-κB(NF-κB)/NF-κB抑制因子(IκB)mRNA表达。结果:模型组细胞MDA,TM,VEGF分泌水平以及NF-κB mRNA表达均显著升高(P<0.01),SOD活性显著降低(P<0.01);FGV保护组可显著降低模型细胞增殖抑制率,MDA,TM,VEGF分泌水平以及NF-κB mRNA表达(P<0.05,P<0.01),提高SOD活性(P<0.01)。结论:FGV具有明显的抗氧化损伤及保护HUVECs的作用,其机制可通过对NF-κB/IκB信号途径的调节。Objective： To study the effects of total flavones of Galium verum （FGV） on protecting in human umbilical vein endothelial cells （HUVECs） injured by hydrogen dioxide （H202 ）. Method： HUVECs were cultured in vitro. An H202injured model was established, FGV at dose of 12.5, 25, 50 mg·L-1was cultured with the cells for 24 h before the model establishment. HUVECs were randomly divided into five groups ： normal control group, model group, and FGV group （12.5, 25, 50 mg·L-1）. Cell viability was measured by methyl thiazolyl tetrazolium （MTT） assay. The activity of superoxide dismutase （SOD） and the secretion levels of malondialdehyde （MDA） were determined by spectrophotometry. The secretion levels of thrombomodulin （TM） and vascular endothelial growth factor （VEGF） were determined using ELISA. The expression levels of NF-kappa B （NF-κB） mRNA and NF-kappa B inhibitor （IκB） mRNA were measured by RT-PCR. Result： The content of MDA, TM, VEGF and NF-κB mRNA were markedly increased in model group （P 〈0.01 ）. Compared to model group, FGV group could reduce the proliferation inhibition of HUVECs, depress the content of MDA, improve the activity of SOD, decrease the secretion levels of TM and VEGF, down-regulated the expression of NF-KB mRNA （P 〈 0.05,P 〈0.01 ）. Conclusion： FGV has a protective effect on the HUVECs impairment induced by H202, and the potential mechanism of action may be associated with down-regulating the expression of NF-κB mRNA.

关 键 词：蓬子菜总黄酮 人脐静脉内皮细胞 过氧化氢 抗氧化 核因子-κB/核因子-κB抑制因子 

分 类 号：R285.5[医药卫生—中药学]