从TLRs介导的炎症信号通路研究补肾方的抗肝损伤机制  

Study on the Anti-hepatic Damage Mechanism of Bushen Recipe from the Inflammatory Signaling Pathway Mediated by TLRs

作　　者：赵彬彬[1] 汪蓉 姜煜资 张莹[1] 高月求[1] 陈建杰[1] 王灵台[1] 聂红明[1] ZHAO Bin-bin;WANG Rong;JIANG Yu-Zi;NIE Hong-ming(Shanghai University of Traditional Chinese Medicine ( Shanghai,200021 )China)

机构地区：[1]上海中医药大学附属曙光医院,上海200021 [2]上海市浦东新区中医医院

出　　处：《中西医结合肝病杂志》2019年第3期236-239,I0004,共5页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：国家自然科学基金(No.81373618,81001492);第六批全国老中医药专家学术经验继承项目(No.国中医药人教发[2017]29号)

摘　　要：目的:从TLRs介导的炎症信号通路探讨补肾方的抗肝损伤机制。方法:将42只SPF级健康成年C57BL小鼠,雌雄各半,随机分为正常组、模型组、补肾全方组和拆方组(补肾阴组、补肾阳组、清化湿热组、引经药组)。每天给药1次,连续给药7天,第7天给药后4h,除正常组外,均给予刀豆蛋白A 15mg/kg尾静脉注射,注射后18h摘眼球处死老鼠。检测肝功能,肝组织HE染色,ELISA法检测外周血相关炎症介质,Western Blot法检测TLR3/4/9相关信号通路链中关键蛋白。结果:肝功能指标中,补肾全方组的ALT、AST均显著下降(P<0.05)、ChE显著上升(P<0.05);补肾阳组在AST中下降明显(P<0.05),补肾阴组在ALT中下降明显(P<0.05)。肝组织HE染色表明,各组均具有一定的改善肝组织炎症坏死的作用,与模型组相比,补肾全组、补肾阳组中坏死区域的面积显著减小。外周血主要炎症介质结果表明,补肾全方组的IL-6、IL-1、TNF-α、TNF-β均显著下降(P<0.05),补肾阳组的TNF-β和IL-1显著下降(P<0.05),而补肾s阴组的TNF-α、TNF-β和IL-6显著下降(P<0.05)。Western Blot检测结果表明,ConA处理后TLR3/4/9相关信号通路链中的关键蛋白如TBK1、IRF3、TLR9、TLR4、TLR3、TRAF6、NF-κB、TIRP、myd88的表达均增加,补肾全方及其拆方处理后,TBK1、IRF3、TLR9、TLR4、TLR3、NF-kB、MyD88在补肾全组中表达减少,TLR9、TLR4、TRAF6、NF-κB、MyD88在补肾阳组中表达减少,TLR9、TLR3、TRAF6、NF-kB在补肾阴组中表达减少,TRAF6在清化湿热组中表达减少。结论:补肾方能通过影响TLRs介导的炎症信号通路,下调炎症介质,发挥其抗肝损伤作用。Objective:To investigate the mechanism of Bushen Recipe against liver injury from TLRs-mediated inflammatory signaling pathway.Methods:Forty-two SPF healthy adult C57 BL mice, male and female, were randomly divided into normal group, model group, Bushenquanfang group and splitting group(Benyanyin group, Bushenyang group, Qinghua damp heat group, Jingjing group).Medicine group).The drug was administered once a day for 7 days, and 4 hours after the 7 th day, except for the normal group, the concanavalin A 15 mg/kg was administered to the tail vein, and the eyeball was sacrificed 18 h after the injection.Liver function was detected, HE staining of liver tissue, peripheral blood-related inflammatory mediators were detected by ELISA, and key proteins in TLR3/4/9-related signaling pathway chain were detected by Western Blot. Results:In the liver function index, the ALT and AST of the tonifying kidney group were significantly decreased(P<0.05) and the ChE was significantly increased(P<0.05).There was no significant decrease in TBIL and DBIL in each drug-administered group(P<0.05).The tonifying kidney group decreased significantly in AST, and the tonifying kidney group decreased significantly in ALT(P<0.05).HE staining of liver tissue showed that each group had a certain effect on improving inflammation and necrosis of liver tissue, but the area of necrotic area in Bushenquan group and Bushenyang group was significantly reduced compared with the model group.The results of main inflammatory mediators in peripheral blood showed that IL-6, IL-1, TNF-α and TNF-β in the kidney group were significantly decreased(P<0.05), and TNF-β and IL-1 in the Bushenyang group were significantly decreased(P<0.05), while TNF-α, TNF-β and IL-6 in the Bushenyin group decreased significantly(P<0.05).Western Blot assay showed that the expression of key proteins such as TBK1, IRF3, TLR9, TLR4, TLR3, TRAF6, NF-kB, TIRP and myd88 in the TLR3/4/9-related signaling pathway chain after ConA treatment increased.After treatment with the disassemble

关 键 词：TLR信号通路 补肾方 肝损伤机制 

分 类 号：R285.5[医药卫生—中药学]