柔肝固肠方改善慢性酒精性肝损伤大鼠肠道通透性的机制研究

摘要

Objective:From the intestinal tight junction and adherens junction pathway to explore mechanism research of Rougan-guchang Decoction on improving chronic alcoholic liver injury and intestinal permeability changes.Methods:The rats were given Lieber-Decarli liquid diet daily for 6 weeks to duplicate the alcoholic liver injury model.Thirty male Sprague-Dawley (SD)rats were randomly divided into control liquid diet,control group (control group,n =1 0)and ethanol liquid diet model group (n =20).Model rats were randomly divided into the model group (n =1 0),Rouganguchang Decoction group (n =1 0)the fourth weeks.The rats were given decoction or distilled water from the fourth week to the sixth weekend.Each rat was given 1 0mg/kg en-dotoxin (LPS)by gavage 3.5h before being sacrificed.The following indexes were detected:1 )The ALT、AST activity of serum were detected by biochemical method;2 )LPS content in the portal blood was detected by biochemical method;3 )Pathological changes in intestinal tissues were observed by electron microscope;4)The protein and mRNA expresses of ZO-1 and Occludin were detected by Western blotting and real-time PCR respectively;5 )The protein of beta-Catenin and E-Cadherin were detected by Western blotting.Results:1 )Compared with that of the model group rats,the activity of AST in drug group decreased significantly (P<0.05);2)The plasma endotoxin levels of Rouganguchang Decoction decreased significantly compared with that of the model group rats;3)Rouganguchang Decoction could improve the intestinal mucosa connection structure of the surface epithelial cells of model rats;4)Compared with those of the model group rats,the protein and mRNA expressions of ZO-1 and occludin,as well as the protein expressions of beta-Catenin and E-Cadherin all increased.Conclusion:Rouganguchang Decoction can significantly im-prove chronic alcoholic liver injury and intestinal permeability changes,the mechanism of that was related to protection of intestinal epithelial tight junctions and adhesion junctions.%目的：从肠道紧密连接及黏附连接角度探讨柔肝固肠方改善慢性酒精性肝损伤大鼠肠道通透性的作用机制。方法：Lieber-DeCarli酒精液体饲料饲养6周诱导大鼠慢性酒精性肝损伤模型。30只SD雄性大鼠，随机分无酒精液体饲料对照组（对照组，n＝10）和酒精液体饲料造模组（n＝20），造模第4周将造模组大鼠随机分模型组（n＝10），柔肝固肠方组（n＝10），并开始灌胃给柔肝固肠方或蒸馏水直至6周末，取材前3.5 h各组以10 mg／kg的内毒素（LPS）灌胃。取材后检测：1）血清AST、ALT活性变化；2）通过门脉血浆内毒素含量测定判断小肠通透性变化；3）小肠组织电镜超微结构观察；4）小肠紧密连接蛋白ZO-1、Occludin蛋白与mRNA表达变化；5）小肠黏附连接蛋白β-Catenin和E-Cadherin蛋白表达。结果：1）柔肝固肠方可显著降低模型大鼠显著升高的AST水平（P＜0.05）；2）柔肝固肠方可降低模型大鼠显著升高的小肠通透性，柔肝固肠方组血浆内毒素含量显著低于模型组；3）电镜结果显示柔肝固肠方可显著改善Lieber-DeCarli酒精饲料喂养大鼠小肠黏膜表面微绒毛局灶性减少、变短、稀疏，排列不规则，同微绒毛相连的细胞终末网变性模糊等病理改变；4）柔肝固肠方可显著升高酒精饲料喂养肝损伤大鼠小肠组织紧密连接蛋白ZO-1、Occludin蛋白与mRNA表达及黏附连接蛋白β-Catenin和E-Cadherin蛋白表达。结论：柔肝固肠方通过改善肠上皮紧密连接及黏附连接改善慢性酒精性肝损伤大鼠肠道通透性改变。