益气活血复方对Toll样受体4信号转导通路及下游炎症因子的影响

Effects of Yiqi Huoxue Compound Contained Drug Serum on the Expressions of Toll-like Receptor 4 and Downstream Signal Transduction Pathway,and LOX-1,TNF-α,ICAM-1 in the Human Vein Endothelial Cells

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DOI：

中文关键词: 中药含药血清内皮细胞 Toll样受体4 髓样分化因子88 血凝素样氧化低密度脂蛋白受体-1 动脉粥样硬化

英文关键词:Chinese medicine contained serum endothelial cell toll-like receptor-4 MyD88 hemagglutinin low-concentration lipoprotein receptor-1 artherosclerosis

基金项目:国家自然科学基金资助项目(No.81160476)

作者	单位
姜华	延边大学附属医院
姜玉姬	延边大学附属医院

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中文摘要:

目的研究益气活血复方含药血清对脂多糖(lipopolysaccharide,LPS)诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)Toll样受体4(Toll-like receptor4,TLR4)信号转导通路及下游炎症因子血凝素样氧化低密度脂蛋白受体-1(LOX-1)、肿瘤坏死因子(TNF-α)、细胞间黏附分子-1(ICAM-1)的影响,从基因和蛋白水平研究益气活血复方防治动脉粥样硬化(atherosclerosis,AS)的机制。方法含药血清的制备:选择新西兰大耳白兔20只,随机分为4组,即空白对照组、中药高浓度组、中药中浓度组、中药低浓度组,每组5只。正常组以生理盐水灌胃;中药高浓度组、中药中浓度组、中药低浓度组分别以高、中、低浓度益气活血复方灌胃,连续灌胃7天。在末次灌胃给药2h后,心脏取血,离心后分离血清。体外培养HUVECs,随机分为6组,即空白对照组、模型组、西药对照组、中药高、中、低浓度组,用LPS刺激2h后,分别加入高、中、低浓度益气活血复方含药血清干预,24h后收集细胞,用荧光定量PCR方法测定TLR4、骨髓样分化因子88(MyD88)、肿瘤坏死因子受体相关因子-6(TRAF-6)、Toll样受体相关分子(TRAM)、Toll样受体相关的干扰素活化因子(TRIF)、核转录因子-κB(NF-κB)、LOX-1、TNF-α及ICAM-1的基因表达,用Western blot法分别测定TLR4、MyD88、TRAF-6、LOX-1的蛋白表达。结果用LPS刺激HUVECs后,引起TLR4、MyD88、TRAF-6、TRAM、TRIF、NF-κB、LOX-1、TNF-α及ICAM-1的高表达,与空白对照组比较,差异有统计学意义(P<0.01),而用益气活血复方含药血清干预以后显著抑制TLR4、MyD88、TRAF-6、NF-κB、LOX-1、TNF-α及ICAM-1的高表达,与模型组比较,差异有统计学意义(P<0.05);对TRAM和TRIF无抑制作用。结论益气活血复方可抑制TLR4信号转导通路及LOX-1、TNF-α及ICAM-1的表达,这可能是其治疗各种免疫炎症性疾病及防治动脉粥样硬化作用的机制之一。

英文摘要:

Objective To investigate the effects of Yiqi Huoxue Compound(YHC) contained drug serum on the expressions of TLR4 and its downstream signal transduction pathway and LOX-1,TNF-α,ICAM-1 in human umbilical vein endothelial cells(HUVECs),and to study its possible anti-atherosclerosis(AS) mechanism.Methods Twenty New Zealand rabbits were divided into 4 groups in random,i.e.,the normal control group,the high concentration group,the middle concentration group,and the low concentration group,5 in each group.Normal saline was given to rabbits in the normal control group by gastrogavage.High,middle,and low concentration of YHC was respectively given to rabbits in the rest 3 groups by gastrogavage for 7 successive days.The blood was withdrawn from the heart 2 h after the last gastrogavage.The serum was isolated after centrifuge.HUVECs was in vitro cultured and then randomly divided into 6 groups,i.e.,the normal control group,the model group,the Western medicine control group,the high,middle,and low YHC groups.HUVECs were stimulated with LPS for 2 h,and then treated with high,middle,and low YHC contained serum.HUVECs were collected 24 h later.The gene expressions of TLR4,MyD88,TRAF-6,TRAM,TRIF,NF-κB,LOX-1,TNF-α,and ICAM-1 were detected using fluorescent quantitative PCR.The protein expressions of TLR4,MyD88,TRAF-6,and LOX-1 were determined using Western blot method.Results After HUVECs were stimulated by LPS,the expressions of TLR4,MyD88,TRAF-6,TRAM,TRIF,NF-κB,LOX-1,TNF-α,and ICAM-1 were enhanced,showing statistical difference when compared with the vehicle control group(P<0.01).YHC contained serum significantly decreased the higher expressions of TLR4,MyD88,TRAF-6,NF-κB,LOX-1,TNF-α,and ICAM-1,showing statistical difference when compared with the model group(P<0.05).Conclusions YHC could inhibit the TLR signal transduction pathway and the expressions of LOX-1,TNF-α,and ICAM-1.These might be one of the mechanisms for treating various immune inflammatory diseases and preventing AS.

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