基于肠-肝-脑轴探讨益木脑液灌肠对肝性脑病大鼠TLR4/NF-κB/NLRP3通路的影响  

Dicussion on the effects of Yimu Naoye enema on TLR4-NF-κB-NLRP3 pathway in hepatic encephalopathy rats based on the enteric-hepato-brain axis

作　　者：高婵婵 张海博 魏海梁 闫曙光[1] 李京涛 周振国 郭英君[4] 常占杰 GAO Chan-chan;ZHANG Hai-bo;WEI Hai-liang;YAN Shu-guang;LI Jing-tao;ZHOU Zhen-guo;GUO Ying-jun;CHANG Zhan-jie(The First Clinical School of Shaanxi University of Chinese Medicine,Xianyang 712046,China;Institute of Medical Sciences,Dalian Medical University,Dalian 116000,China;Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China;People’s Hospital of Ningxia Hui Autonomous Region,Yinchuan 750001,China)

机构地区：[1]陕西中医药大学第一临床医学院,咸阳712046 [2]大连医科大学医学科学研究院,大连116000 [3]陕西中医药大学附属医院,咸阳712000 [4]宁夏回族自治区人民医院,银川750001

出　　处：《中华中医药杂志》2023年第6期2599-2604,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82174330);陕西省科技厅科研基金项目(No.2020ZDLSF05-15,No.2019SF-309);陕西中医药大学学科创新团队建设项目(No.2019-YL05)。

摘　　要：目的:观察中药益木脑液灌肠对肝性脑病大鼠的治疗作用,并探讨其对TLR4/NF-κB/NLRP3通路表达的影响。方法:72只大鼠随机分为正常组,模型组,益木脑液高、中、低剂量组,乳果糖组,每组12只。除正常组外,其余大鼠均给予0.25 mL/100 g的CCl4溶液腹腔注射,每周2次,共6周,制备大鼠肝损伤模型。6周后给予300 mg/kg的硫代乙酰胺溶液隔日腹腔注射,共2次,建立大鼠肝性脑病模型。造模成功后各组大鼠给予相应剂量的药物灌肠,给药10 d后进行神经功能反射评分、行为学检测(水迷宫实验、旷场实验),取血清、血浆及肝脏组织样本。检测各组大鼠ALT、AST、血浆氨含量;ELISA法检测血清白介素6(IL-6)、肿瘤坏死因子-α(TNF-α)、脂多糖(LPS)水平;HE染色观察肝脏病理变化;免疫组化观察肝组织TLR4、NF-κB、NLRP3蛋白含量;Western blot法检测肝脏TLR4、NF-κB、NLRP3、Caspase1蛋白的表达。结果:与模型组比较,益木脑液中、高剂量组能显著改善肝性脑病大鼠神经功能反射评分,提高大鼠行为学水平,降低ALT、AST、血浆氨、IL-6、TNF-α、LPS含量,抑制肝脏TLR4、NF-κB、NLRP3、Caspase1蛋白表达(P<0.05,P<0.01)。结论:益木脑液灌肠可改善大鼠肝脏炎症,减轻炎症损伤,从而治疗肝性脑病。其机制可能与抑制TLR4/NF-κB/NLRP3信号通路的活化进而调节肝脏非可控性炎症有关。Objective:To observe the therapeutic effect of Chinese medicine Yimu Naoye enema on hepatic encephalopathy rats,and explore its effect on the expression of TLR4/NF-κB/NLRP3 pathway.Methods:A total of 72 rats were randomly divided into normal group,model group,Yimu Naoye high-dose group,medium-dose group,low-dose group,milk Fructose group,12 rats in each group.Except for the normal group,the remaining 60 rats were injected intraperitoneally with 0.25 mL/100 g CCl 4 solution twice a week for 6 weeks to prepare rat liver injury models.Six weeks later,300 mg/kg of thioacetamide solution was injected intraperitoneally every other day for 2 times to establish a rat hepatic encephalopathy model.After successful modeling,each group was given corresponding doses of drug enema,neurological reflex scores and behavioral tests(water maze test,open field test)were performed 10 days after administration,and serum,plasma,liver tissue samples were taken.Detect liver function and plasma ammonia content of rats in each group;ELISA method to detect interleukin 6(IL-6),tumor necrosis factor-α(TNF-α),lipopolysaccharide LPS levels;HE staining to observe liver pathological changes;immunohistochemical observation the contents of TLR4,NF-κB,and NLRP3 in liver tissue;Western blot method was used to detect the protein expression of the TLR4,NF-κB,NLRP3 and Caspase1 in the liver.Results:Compared with the model group,Yimu Naoye medium-dose group,high-dose group can significantly improve the neurological reflex score of hepatic encephalopathy rats,improve the behavioral level of rats,and reduce the content of ALT,AST,plasma ammonia,IL-6,TNF-α,and LPS after enema;Inhibit the protein expression of liver TLR4,NF-κB,NLRP3,Caspase1(P<0.05,P<0.01).Conclusion:Yimu Naoye enema can improve liver inflammation and reduce inflammation damage in rats,thereby treating hepatic encephalopathy.The mechanism may be related to inhibiting the activation of TLR4/NF-κB/NLRP3 signaling pathway and regulating uncontrollable liver inflammation.

关 键 词：肝性脑病 益木脑液 TOLL样受体4 核因子-ΚB NOD样受体热蛋白结构域相关蛋白3 肝脏炎症 

分 类 号：R285.5[医药卫生—中药学]