| Objective:Using ?-Naphthylisothiocyanate(ANIT)to make a model of subacute intrahepatic cholestasis in neonatal rats,discusses the protective mechanism of lactobacillus rhamnosus GG(ATCC 53103)on liver function in in neonatal rats with cholestasis and liver fibrosis.Methods:Healthy male Spragne-Dawlay rats,aged 4 weeks,reared at animal experimental center in the Third Hospital of Hebei Medical University.80 Spragne-Dawlay rats were randomly divided into 5 groups:There were 16 in the control group(group A),16 in the disease observation group(group B1),16 in the ursodeoxycholic acid intervention group(group B2),16 in the lactobacillus rhamnosus GG intervention group(group B3)and 16 in the combined intervention group(group B4).Rats in the model group were given ANIT gavage to induce subacute intrahepatic cholestasis,and rats in group A were given sesame oil by gavage.Different interventions were carried out after successful modeling:no drug treatment was given in group B1;B2 group was given UDCA 100 mg / kg in subacute cholestasis model rats,1/day;B3 group was given 2.5ml lactobacillus rhamnii GG(LGG)in the subacute cholestasis model,1/day;B4 group was given UDCA 100mg/kg and lactobacillus rhamnii GG 2.5ml in the subacute cholestasis model mice,1/day;Continuous irrigation for 23 days.Rat blood and liver samples were collected according to each observation time node,the corresponding biochemical indexes were detected,such as alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),direct bilirubin(DBIL),and bile acid(TBA);Liver fibrosis indicators such as hyaluronic acid(HA),type III procollagen(PCIII),type IV collagen(c IV),laminin(LN)and liver pathology.SPSS23.0 software was used for statistical analysis.Results:1.SD Rats irrigation take alpha naphthyl isothiocyanates(alpha Naphthylisothiocyanate,ANIT)after the mental tired,decreased activity,poor appetite,the skin is dark yellow,hair messy lose luster,deep yellow color of urine.Pathology of the liver showed that the structure of the liver tissue was damaged,accompanied by inflammatory cell infiltration in the portal area,bile duct stricture,cholestasis and fibrous connective tissue hyperplasia were observed,and a subacute cholestasis rat model was established.2.Blood biochemical indexes such as TBIL,DBIL,TBA,ALP,ALT,AST,and liver fibrosis indexes such as HA,LN,and c IV in the subacute cholestasis model rats showed significant changes compared with the healthy control rats(P < 0.05),and the liver pathology showed obvious changes of cholestasis and liver fibrosis.3.After different interventions,it was found that group B2,B3 and B4 all improved the general state of subacute cholestasis model rats,such as spirit,appetite and activity,while group B3 showed more obvious improvement.4.In terms of improving the metabolic indexes of bilirubin,such as serum total bilirubin,direct bilirubin and bile acid,on the 2nd day after the intervention in the subacute cholestasis model rats,the above indexes in the blood of group B1,B2,B3 and B4 were all higher than those in group A(P < 0.05);On day 9,the rats in group B3 decreased significantly compared with group B1(P < 0.05);On day 16,the above indexes of B3 and B4 groups were different from those of B1 group(P < 0.05);On the 23 rd day,there were differences between group B2,B3,B4 and B1,but the B4 group showed significant difference(P < 0.05).5.In terms of improving liver function indexes such as alanine aminotransferase and aspartate aminotransferase,the blood indexes of group B1,B2,B3 and B4 were all higher than those of group A on the second day after intervention in the subacute cholestasis model rats(P < 0.05);On day 9,the rats in group B3 decreased significantly compared with group B1(P < 0.05);On day 16,there were differences between B3 and B4 groups and B1 groups(P < 0.05);On the 23 rd day,there was a significant difference between group B2,B3 and B4 and group B1(P < 0.05);In terms of improving liver fibrosis indexes such as hyaluronic acid,laminin and type IV collagen,the blood indexes in group B1,B2,B3 and B4 were all higher than those in group A on the 2nd day after intervention in the subacute cholestasis model rats(P < 0.05);On day 9,the rats in group B3 decreased significantly compared with group B1(P < 0.05);On day 16,there were differences between B3 and B4 groups and B1 groups(P < 0.05);On the 23 rd day,there were differences between group B2,B3,B4 and B1,but the B4 group showed significant difference(P < 0.05).6.Pathological examination of the liver under 200-fold light microscope showed that the model group of rats all had hepatocyte degeneration and necrosis,biliary cholestasis and liver fibrosis.7.In terms of the above-mentioned improvement of bilirubin metabolism,liver function and liver fibrosis indexes,the intervention measures of B2,B3 and B4 groups all showed different degrees of improvement,and the B4 group had the best effect.Conclusions:1.ANIT solution was administered to SD rats to establish an animal model of hepatic fibrosis in subacute intrahepatic cholestasis.2.Different interventions were used in the subacute cholestasis model rats,and it was found that all the interventions could improve the general state of the model rats,especially lactobacillus rhamnosus GG(ATCC 53103).3.For the subacute cholestasis model mice,different intervention measures were used to improve the bilirubin metabolism and liver function indexes.It was found that both lactobacillus rhamnosus GG and ursodeoxycholic acid had better intervention effects,especially the latter,indicating that both of the two interventions had better cholagogic and liver protective effects.4.Lactobacillus rhamnosus GG combined with ursodeoxycholic acid has a good therapeutic effect on the improvement of liver fibrosis in subacute cholestasis model rats,which can effectively protect the liver function during cholestasis and resist the liver fibrosis caused by cholestasis.5.The therapeutic effect of lactobacillus rhamnosus GG combined with ursodeoxycholic acid may be related to the reconstruction of intestinal microecological balance,the maintenance of intestinal mucosal barrier function and the change of intestinal bile acid metabolism during cholestasis. |
PDF Full Text Request