基于细胞骨架重塑促进肠上皮细胞AJC组装探讨揉腹法治疗非酒精性脂肪肝病的作用机制

Effect of sputum abdomen therapy on non-alcoholic fatty liver disease based on cytoskeletal remodeling to promote AJC assembly of intestinal epithelial cells

目的探究揉腹法对高脂饮食诱导的非酒精性脂肪肝病(NAFLD)大鼠的肝脏保护作用及其部分作用机制。方法选取43只6~8周龄雄性SD大鼠,随机分为两组:正常对照组(10只,A组),造模组(33只)。造模组大鼠给予高脂饲料饲养建造非酒精性脂肪肝病模型,将造模成功的大鼠均分为模型组(B组)、揉腹法组(C组)和药物治疗组(D组)。治疗4 w后,对比B、C、D组大鼠体重、肝湿重变化,酶联免疫吸附试验(ELISA)检测大鼠血清中天冬氨酸转移酶(AST)、血清游离脂肪酸(FFA)、丙氨酸氨基转移酶(ALT)和三酰甘油(TG)水平,采用苏木素-伊红(HE)染色观察肝脏和回肠组织病理改变,异硫氰酸荧光素-葡萄糖(FITC-Dextran)法观察肠道黏膜通透性,免疫组化和Western印迹检测肠黏膜组织中闭合小环蛋白(ZO)-1,咬合蛋白(Occludin),上皮型钙黏蛋白(E-cadherin),β-连环蛋白(catenin),闭合蛋白(Claudin)1表达。结果高脂饲料饲养10 w后,模型组大鼠体重和肝湿重明显增加,肝脏组织病理发生明显改变,提示造模成功。重新分组治疗4 w后,与B组相比,C组和D组大鼠体重及肝湿重明显降低(P<0.05),肝组织和肠组织病理损伤明显改善;C和D组肠黏膜组织ZO-1、Occludin、E-cadherin、β-catenin、Claudin1较B组明显增高(P<0.05);C和D组大鼠肠袋外漏出FITC-Dextran含量较B组明显降低(P<0.05)。结论揉腹法可降低NAFLD大鼠的体重和肝湿重,改善肝组织和肠组织病理损伤,其作用机制是通过刺激细胞骨架重塑促进肠上皮细胞顶端连接复合体(AJC)组装,改善肠上皮屏障和通透性,从而发挥肝脏保护作用。

Objective To explore the protective effect of sputum abdomen on the liver of non-alcoholic fatty liver disease(NAFLD)induced by high fat diet and its partial mechanism.Methods Forty-three male Sprague-Dawley rats aged 6~8 weeks were randomly divided into two groups:normal control group(10 rats,group A)and model group(33 rats).Model rats were fed with high-fat diet to build a model of non-alcoholic fatty liver disease.Rats with successful modeling were divided into model group(group B),abdominal abdomen group(group C)and drug treatment group(group D).After 4 weeks of treatment,the body weight and liver wet weight of rats in group B,C and D were compared.The serum aspartate transferase(AST),serum free fatty acid(FFA)and alanine amino group were detected by ELISA.The levels of transferase(ALT)and triglyceride(TG)were observed by HE staining.The intestinal mucosal permeability was observed by FITC-Dextran method.ZO-1,Occludin,E-cadherin,β-catenin,Claudin1 protein expression in intestinal mucosa were detected by immunohistochemistry and Western blot.Results After 10 weeks of high-fat diet,the body weight and liver wet weight of the model group increased significantly,and the pathological changes of liver tissue changed significantly,which suggested that the model was successful.After 4 weeks of regrouping,compared with group B,the body weight and liver wet weight were significantly lower,and the pathological damage of liver tissue and intestinal tissue were significantly improved,expressions of ZO-1,Occludin,E-cadherin,β-catenin,Claudin1 in intestinal mucosa tissue were significantly higher,the content of FITC-Dextran leaked out from the intestines were significantly reduced in group C and group D(all P<0.05).Conclusions The sputum method could reduce the body weight and liver wet weight of NAFLD rats and improve the pathological damage of liver tissue and intestinal tissue.The mechanism of action is to promote the assembly of opical junctional complex(AJC)in intestinal epithelial cells by stimulating cytoskeletal remodeling and improve intestinal epithelial barrier and permeability to play a role in liver protection.