海珠益肝加味方对慢性免疫性肝损伤小鼠LPS/TLR4信号途径的影响  

Effects of Haizhu Yigan Decoction on LPS/TLR4 signaling pathway in mice with chronic immunological liver injury

作　　者：崔翔[1,2] 于慧杰 盛国光 

机构地区：[1]湖北中医药大学,武汉430065 [2]安康市中医医院,陕西安康725000 [3]淄博市中西医结合医院,山东淄博255026 [4]湖北省中医医院肝病研究所,武汉430061

出　　处：《中华中医药杂志》2016年第12期5192-5195,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家中医临床研究基地业务建设基金项目(No.JDZX2012049);湖北省科技厅科技计划项目项目(No.2012DGA12008)~~

摘　　要：目的:研究海珠益肝加味方对刀豆蛋白(Con A)诱导的慢性免疫肝损伤小鼠脂多糖/Toll样受体4(LPS/TLR4)信号途径的影响。方法:C57BL/6小鼠48只,随机均分为4组,空白对照组、模型组、甘草酸二铵组、海珠益肝加味方组,采用尾静脉Con A注射法复制模型。造模成功后,空白对照组、模型组予20m L/kg蒸馏水灌胃,甘草酸二铵组予甘草酸二铵0.68mg·kg-1·d-1灌胃,海珠益肝加味方组予海珠益肝加味方15.17g·kg-1·d-1灌胃。干预4周后,采集血清检测血清丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)、总胆红素(TBi L);鲎试剂显色基质法测定脂多糖(LPS);ELISA法检测肿瘤坏死因子(TNF-α)、白介素-6(IL-6);Western Blot检测肝脏组织TLR4,My D88,核因子-κB(NF-κB)蛋白表达。结果:与模型组比较,海珠益肝加味方组小鼠ALT、AST、TBi L水平显著下降(P<0.05,P<0.01);与模型组和甘草酸二铵组比较,海珠益肝加味方干预组LPS水平显著降低(P<0.05);与模型组比较,海珠益肝加味方干预组TNF-α、IL-6水平均明显下降(P<0.05);与模型组和甘草酸二铵组比较,海珠益肝加味方组小鼠肝组织TLR4、My D88、NF-κB的蛋白表达显著下调(P<0.05,P<0.01)。结论:海珠益肝加味方保护肝脏的作用机制与抑制TLR4-My D88-NF-κB信号通路有关。Objective： To investigate the effect of Haizhu Yigan Decoction on LPS/TLR4 signaling pathway in mice with chronic immunological liver injury. Methods： Forty-eight C57BL/6 mice were randomly divided into 4 groups, including control group, model group, diammonium glycyrrhizinate group and modified Haizhu Yigan Decoction group. After the models were successfully established, rats in control group and model group received gavage administration with 0.9% Na Cl, rats in diammonium glycyrrhizinate group received gavage administration with 0.68mg·kg-1·d-1 of diammonium glycyrrhizinate, and rats in Haizhu Yigan Decoction group received gavage administration with modified Haizhu Yigan Decoction 15.17g·kg-1·d-1. Serum alanine aminotransferase（ALT）, aspartate aminotransferase（AST）, and total bilirubin（TBi L） were measured after 4 weeks of intervention. The level of LPS was tested by chromogenic method of limulus reagent, and the level of TNF-α and IL-6 was tested by ELISA. The protein expressions of TLR4, My D88 and NF-κB in liver tissues were detected by Western Blot. Results： Compared with model group, the levels of ALT, AST and TBi L in mice treated with Haizhu Yigan Decoction were significantlydecreased（P〈0.05, P〈0.01）. Compared with model group and diammonium glycyrrhizinate intervention group, LPS level of mice in Haizhu Yigan Decoction group decreased significantly（P〈0.05）. Compared with model group, the levels of TNF-α and IL-6 of mice in Haizhu Yigan Decoction group decreased significantly（P〈0.05）. Compared with model group and diammonium glycyrrhizinate group, the protein expression of TLR4, My D88 and NF-κB in liver tissue of mice treated with modified Haizhu Yigan Decoction decreased significantly（P〈0.05, P〈0.01）. Conclusion： The protective effect of Haizhu Yigan Decoction on liver was related to the inhibition of TLR4-My D88-NF-κB signaling pathway.

关 键 词：海珠益肝加味方 刀豆蛋白A 慢性免疫性肝损伤 肠源性内毒素血症 脂多糖/Toll样受体4信号途径 

分 类 号：R285.5[医药卫生—中药学]