CAJ | 학술논문

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。
목적：종장도긴밀련접급점부련접각도탐토유간고장방개선만성주정성간손상대서장도통투성적작용궤제。방법：Lieber-DeCarli주정액체사료사양6주유도대서만성주정성간손상모형。30지SD웅성대서，수궤분무주정액체사료대조조（대조조，n＝10）화주정액체사료조모조（n＝20），조모제4주장조모조대서수궤분모형조（n＝10），유간고장방조（n＝10），병개시관위급유간고장방혹증류수직지6주말，취재전3．5 h각조이10 mg／kg적내독소（LPS）관위。취재후검측：1）혈청AST、ALT활성변화；2）통과문맥혈장내독소함량측정판단소장통투성변화；3）소장조직전경초미결구관찰；4）소장긴밀련접단백ZO-1、Occludin단백여mRNA표체변화；5）소장점부련접단백β-Catenin화E-Cadherin단백표체。결과：1）유간고장방가현저강저모형대서현저승고적AST수평（P＜0．05）；2）유간고장방가강저모형대서현저승고적소장통투성，유간고장방조혈장내독소함량현저저우모형조；3）전경결과현시유간고장방가현저개선Lieber-DeCarli주정사료위양대서소장점막표면미융모국조성감소、변단、희소，배렬불규칙，동미융모상련적세포종말망변성모호등병리개변；4）유간고장방가현저승고주정사료위양간손상대서소장조직긴밀련접단백ZO-1、Occludin단백여mRNA표체급점부련접단백β-Catenin화E-Cadherin단백표체。결론：유간고장방통과개선장상피긴밀련접급점부련접개선만성주정성간손상대서장도통투성개변。
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.