| The pathogenesis of nonalcoholic fatty liver disease(NAFLD)has not yet been fully elucidated.The "intestinal-hepatic axis" theory has become a hot topic in the study of liver diseases in recent years,and the destruction of the intestinal mucosal barrier has attracted everyone's attention.It is considered that intestinal flora-induced endotoxemia(IETM)may play an important role in the pathogenesis of NAFLD due to dysbacteriosis,increased intestinal mucosal permeability and bacterial translocation.Berberine is an isoquinoline alkaloid extracted from Coptis chinensis and has strong anti-inflammatory and immunomodulatory effects.Studies have shown that berberine can reduce the damage caused by chronic stress of the intestinal mucosa;can reduce the expression of inflammatory mediators in severe abdominal infection or sepsis,thereby improving the damaged intestinal mucosa barrier,reducing intestinal permeability,We speculate that berberine hydrochloride may play a certain role in the treatment of NAFLD by improving the intestinal mucosal barrier function.Objective: To establish a rat model of non-alcoholic fatty liver disease(NAFLD)to study the effect of berberine hydrochloride on the intestinal mucosal barrier of NAFLD rats by regulating the intestinal immune barrier,and provide a theoretical basis for berberine hydrochloride to treat NAFLD.Methods:Thirty-three male SD rats(200gą10g)were randomly divided into 3 groups: normal control group(group N),high-fat model group(group M),and Berberine hydrochloride group(group B).Group N was fed with common diets,group M and group B were fed with high-fat diets,and after 12 weeks,one group was sacrificed.HE staining was used to determine the degree of fat deposition in hepatocytes.In Group B,150 mg/(kg.d)berberine hydrochloride was intragastrically administered.The same amount of salinewas given to the N group and the M group.The dose was adjusted according to the change of the body weight of the rats.After continuous gavage for 4weeks,the anesthesia was reduced.Arterial blood.Alanine Aminotransferase(ALT),Aspartate Aminotransferase(AST),Triglyceride(TG),Total Cholesterol(TC)were detected by an automatic biochemical analyzer.Serum endotoxin levels were determined using a limulus test.Liver tissue specimens were obtained and HE pathological changes were observed by HE staining.Serum interleukin-1?(IL-1?),interleukin-18(IL-18),Tumor necrosis factor-?(TNF-?)were measured by ELISA.About 2 cm away from the ileocecal valve,the segments of the small intestine were sectioned,and 2 cm in each segment were taken.RT-PCR was used to detect the expression levels of NOD1,NOD2 and NLRP3.Western blot was used to detect the expression levels of NOD1,NOD2,NLRP3,Caspase-1 and Claudin-4.Results:1.At 12 weeks of feeding,body weights of group M and group B were significantly higher than those of group N(P<0.05).There was no difference in body weight between group B and group M(P>0.05).The results of HE staining showed that fat deposition was observed in the liver cells of rats of group M and group B,which proved that the model was successful.2.At 16 weeks of feeding,after berberine hydrochloride intervention for4 weeks,the body weight of group M rats was significantly higher than that of group N rats(P<0.05);compared with group M rats,group B rats Weight decreased significantly(P<0.05).3.At 16 weeks of feeding,the ALT,AST,TC,and TG in group M were significantly higher than those in group N(P<0.05).Compared with group M,the ALT,AST,TC,and TG in group B were significantly decreased(P<0.05).4.At the 16 th week of feeding,the endotoxin,IL-1?,IL-18 and TNF-? in the M group were significantly higher than those in the N group(P<0.05).Compared with the M group,the B group had endotoxin and IL-1?,IL-18,TNF-? decreased significantly(P<0.05).5.Liver tissue HE staining: The structure of hepatic lobule was clear ingroup N,and hepatocytes did not have steatosis.Compared with group N,hepatic lobule structure was disordered in group M,hepatocytes were swollen,and macrovesicular fat became the main component.The hepatocytes are filled with fat vacuoles of various sizes,and the nuclei are squeezed to the side.After treatment with berberine hydrochloride,the degree of swelling of hepatocytes in group B was reduced,hepatocyte fat was reduced,fat vacuoles were less,and liver cells were arranged more neatly.6.NOD1,NOD2,NLRP3 expression levels in ileum: Rats fed 16 weeks,RT-PCR and Western Blot results showed that the expression of NOD1,NOD2,NLRP3 mRNA and protein were significantly higher in group M than in group N(P<0.05).The expression of NOD1,NOD2,and NLRP3 mRNA and protein was significantly decreased in group B compared with group M(P<0.05).7.Caspase-1 protein expression: After 16 weeks of feeding in rats,Western blot results showed that the amount of Caspase-1 protein in group M was significantly higher than that in group N(P<0.05);Caspase-1 was compared between group B and group M.Protein content decreased significantly(P<0.05).8.Claudin-4 protein expression: At 16 weeks of feeding,the expression of Claudin-4 protein in group M was significantly lower than that in group N(P<0.05);the expression of Claudin-4 protein was significantly increased in group B compared with group M.(P<0.05).Conclusions:1.A high-fat diet can successfully establish a rat model of NAFLD,and the expression level of endotoxin in NAFLD rats is increased.2.Berberine hydrochloride can reduce the body weight of NAFLD rats,improve liver function,reduce blood lipids,and reduce liver steatosis.3.Berberine hydrochloride can reduce the level of endotoxin in rats with NAFLD and reduce the levels of IL-1?,IL-18 and TNF-?.4.Berberine hydrochloride inhibited the expression of NOD1,NOD2,NLRP3 mRNA and NOD1,NOD2,NLRP3,and Caspase-1 proteins in the intestinal mucosal immune barrier.5.Berberine Hydrochloride Increases the Expression of Claudin-4 Protein in Intestinal Mucosa Permeability in NAFLD Rats. |
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