加味人参乌梅汤通过TLR4/MyD88/pNF⁃κBp65信号通路对腹泻大鼠肠黏膜屏障的影响  

Exploring the effect of modified Renshen Wumei Decoction on intestinal mucosal barrier in diarrhea rats based on TLR4/MyD88/pNF⁃κBp65 signaling pathway

作　　者：管志伟[1,2] 赵琼[3] 邱建利[1,2] 徐炎 黄勤挽[3] 周鸿雲 赵君琪 毋颖慧 GUAN Zhiwei;ZHAO Qiong;QIU Jianli;XU Yan;HUANG Qinwan;ZHOU Hongyun;ZHAO Junqi;WU Yinghui(Pediatric Hospital,the First Affiliated Hospital of Henan University of Traditional Chinese Medicine,Zhengzhou 450000,Henan,China;不详)

机构地区：[1]河南中医药大学第一附属医院儿科医院,河南郑州450000 [2]河南中医药大学儿科医学院,河南郑州450046 [3]成都中医药大学药学院,四川成都610075 [4]四川省中医院儿科,四川成都610072

出　　处：《实用医学杂志》2025年第7期944-952,共9页The Journal of Practical Medicine

基　　金：国家自然科学基金面上项目(编号:81674026);河南省科技攻关计划项目(编号:232102311218);河南省中医药科学研究专项(编号:2022JDZX017)。

摘　　要：目的探讨加味人参乌梅汤对TLR4/MyD88/pNF-κBp65信号通路的影响,分析该方修复腹泻大鼠肠黏膜屏障可能的作用机制。方法随机从48只大鼠中选取12只作为空白组(CK),其余36只选用复合方法制作模型。模型制备14 d后随机设置模型组(MC)、西药组(MV)、中药组(MRWD),4组分别给予相应措施干预7 d。ELISA法检测血清DAO、D-Lac、IL-1β、IL-6、IL-10、TNF-α、MUC2、MUC4、MUC6及结肠匀浆SIgA浓度;Western blot、Rt-PCR法检测结肠TLR4、MyD88、pNF-κBp65、Occludin、Claudin-1、ZO-1蛋白和基因表达。结果(1)肠黏膜损伤标志物:与空白组比较,模型组DAO、D-Lac血清含量显著上调(P<0.05);与模型组比较,中、西药组均能显著下调DAO血清含量(P<0.001),中药组可显著下调D-Lac血清含量(P<0.05);中、西药组间DAO、D-Lac血清含量变化差异无统计学意义(P>0.05)。(2)炎症指标:与空白组比较,模型组TLR4、MyD88、pNF-κBp65蛋白、基因表达及IL-1β、IL-6、TNF-α血清含量显著上调(P<0.05),IL-10血清含量显著下调(P<0.05);与模型组比较,中、西药组均能显著下调TLR4、MyD88、pNF-κBp65蛋白、基因表达及IL-1β、IL-6、TNF-α血清含量(P<0.05),并能显著上调IL-10血清含量(P<0.05);中、西药组间TLR4、MyD88、pNF-κBp65蛋白、基因表达及IL-1β、IL-6、IL-10、TNF-α血清含量变化差异无统计学意义(P>0.05)。(3)肠黏膜屏障因子:与空白组比较,模型组MUC2、MUC6、SIgA含量和Claudin-1、ZO-1蛋白、基因表达以及Occludin蛋白表达显著下调(P<0.05);与模型组比较,中、西药均能显著上调MUC2、SIgA含量和Claudin-1、ZO-1蛋白、基因表达(P<0.05),中药组可显著上调MUC6含量和Occludin蛋白表达(P<0.05);中、西药组间MUC2、MUC6、SIgA血清含量变化及Claudin-1、ZO-1基因表达差异无统计学意义(P>0.05),而西药组上调Claudin-1蛋白表达优于中药组(P<0.05),中药组上调ZO-1蛋白表达优于西药组(P<0.05)。结论加味人参乌梅汤�Objective To investigate the effect of Modified Renshen Wumei Decoction on the TLR4/MyD88/pNF⁃κBp65 signaling pathway and elucidate the potential mechanism by which this formula repairs the intestinal mucosal barrier in diarrheal rats.Methods Twelve rats were randomly selected from a total of 48 rats to serve as the blank control group(CK),while the remaining 36 rats were used to establish a disease model via a compound method.After 14 days of model preparation,the rats were randomly divided into three groups:the model group(MC),the western medicine group(MV),and the traditional Chinese medicine group(MRWD).Each of the four groups(including CK)received corresponding interventions for 7 days.The concentrations of serum diamine oxidase(DAO),D⁃lactic acid(D⁃Lac),interleukin⁃1β(IL⁃1β),IL⁃6,IL⁃10,tumor necrosis factor⁃α(TNF⁃α),mucin 2(MUC2),MUC4,MUC6,and colonic homogenate secretory immunoglobulin A(SIgA)were measured using ELISA.Additionally,the protein and gene expressions of colonic toll⁃like receptor 4(TLR4),myeloid differentiation primary response 88(MyD88),phosphorylated nuclear factor kappa⁃light⁃chain⁃enhancer of activated B cells p65(pNF⁃κBp65),occludin,claudin⁃1,and zonula occludens⁃1(ZO⁃1)were analyzed by Western blot and RT⁃PCR.Results(1)Intestinal mucosal injury markers:Compared with the blank group,the serum levels of DAO and D⁃Lac in the model group were significantly increased(P<0.05).Compared with the model group,both the Chinese medicine group and Western medicine group significantly decreased the serum levels of DAO(P<0.001),while the traditional Chinese medicine group also significantly reduced the serum levels of D⁃Lac(P<0.05).There was no significant difference in the changes of DAO and D⁃Lac serum levels between the Chinese medicine group and Western medicine group(P>0.05).(2)Inflammatory indicators:Compared with the blank group,the model group exhibited significant upregulation of TLR4,MyD88,pNF⁃κBp65 protein and gene expression,as well as seru

关 键 词：加味人参乌梅汤 腹泻 TLR4 MYD88 NF-ΚB 肠黏膜屏障 

分 类 号：R285.5[医药卫生—中药学]