| ObjectiveIn the gut-liver axis theory,it is believing that the intestine is closely related liver,and it affects and connects to each other through a variety of media.This study observed the liver damage and intestinal damage in acute cholestasis model,chronic cholestasis model and Tibetan Cold-Liver model.effect of JGST to above model.It is hoped to explain the the characteristics of the gut-liver axis injury and effect of JGST.Providing experimental data for the gut-liver axis research,Tibetan Cold-Liver model research and the Tibetan medicine research.MethodsAcute cholestasis model,chronic cholestasis model,Tibetan Cold-Liver model was bulid and treated by JGST.Observed liver function damage(serum ALT,AST,AKP,TBA)changes;fibrotic lesions shown by Masson staining;Liver fibrosis indicators(αsma,TIM P1,MMP2,and MMP13);intestinal damage(HE staining,expression of intestine ZO-1,Occludin,oxidative damage indicators such as SOD,MDA,GSH,GSSG,GSH/GSSG,small intestinal inflammation factor IL1β;intestinal bile acid receptor FXR.The characteristics of gut-liver axis injury in cholestasis and Tibetan Cold-Liver model were initially investigated and search the effect of JGST.1.Difference between acute cholestasis model and chronic cholestasis model in gut-liver axisAnimals were divided into control group,acute cholestasis group and chronic cholestasis model.The acute cholestasis model group was modeled with a single subcutaneous injection of the cholestasis-inducing agent ANIT for 48 hours to prepare an acute cholestasis model;the chronic cholestasis model group was modeled with subcutaneous injection of ANIT every other day for 14 days to prepare a chronic cholestasis model.Observed the liver injury in acute cholestasis model and chronic cholestasis model and clarify the difference between the two model.Observed the intestinal injury of acute cholestasis model and chronic cholestasis model,and further detected the transcriptional differences of intestinal inflammatory factor IL6 and anti-inflammatory factor IL10,intestinal inflammation-related bile acid receptor TGR5 and intestinal FXR downstream transporter ASBT,clarify the differences in Intestinal damage in between the two models.Observed 25 major bile acids in feces,the differences in fecal bile acid of acute cholestasis and chronic cholestasis model were clarified.2.Regulation of JGST on gut-liver axis injury in chronic cholestasis modelAnimals were divided into control group,chronic cholestasis group,methionine positive group,and JGST group.The chronic cholestasis group was injected subcutaneously with ANIT every other day for 14 days to prepare a chronic cholestasis model.The methionine-positive group was given adenosylmethionine,and the JGST group was given JGST.By observing JGST’s effect on liver injury in chronic cholestasis model,the classic pathway of liver fibrosis TGFβ1-P-Smad2/3,liver bile acid receptors and transporters FXR,TGR5,NTCP and MRP2.The effect of JGST on Liver Injury in chronic cholestasis model was elucidated.By observing JGST’s effect on the intestinal injury of chronic cholestasis model and intestinal inflammation-related bile acid receptor TGR5.The effect of JGST on intestinal injury of chronic cholestasis model was elucidated.3.Differences of gut-liver axis injury between Tibetan Cold-Liver model and chronic cholestasis modelAnimals were divided into control groups,chronic cholestasis model group,the Tibetan Cold-Liver model group,the YinChenHaoTang group,YinChenZhuFuTang group.The chronic cholestasis model group was injected subcutaneously with ANIT every other day for 14 days to prepare a chronic cholestasis model;the Tibetan Cold-Liver model group was intragastrically administered with the Sennae Folium(fan xie ye)every day for 14 days,and the animals were injected subcutaneously with ANIT subcutaneously every other day to build a Yin yellow model.The YinChenHaoTang group was given the YinChenHaoTang as Yang-Huang positive medicine;the YinChenZhuFuTang group was given the YinChenZhuFuTang as Tibetan Cold-Liver positive medicine Yin Chenzhu Fu Decoction group.By observing the liver damage in chronic cholestasis model and Tibetan Cold-Liver model,classic pathway of liver fibrosis TGFβ1-P-Smad2/3,bile acid receptors and transporters FXR,NTCP,MRP2 to clarify the difference between the liver injury in Tibetan Cold-Liver model and chronic cholestasis model.By observing the intestinal damage of chronic cholestasis model and Tibetan Cold-Liver model and the transcription level of intestinal bile acid receptor FXR,the differences between intestinal damage of Tibetan Cold-Liver model and chronic cholestasis model were clarified.By observing the Tibetan Cold-Liver model and chronic cholestasis model,the changes of 25 major bile acid components in serum were explained;the differences in serum bile acid profiles of Tibetan Cold-Liver model and chronic cholestasis model were clarified.4.JGST effect on gut-liver axis injury in Tibetan Cold-Liver modelAnimals were divided into control group,Tibetan Cold-Liver model group,YinChenZhuFuTang group and JGST group.The Tibetan Cold-Liver model group was intragastrically administered with the Sennae Folium(fan xie ye)for 14 days,and AN IT was injected subcutaneously at intervals to build the Tibetan Cold-Liver model.The YinChenZhuFuTang group was treated with YinChenZhuFuTang;JGST group was treated with JGST.By observing JGST’s effect on liver damage,the classic pathway of liver fibrosis,TGFβ1-P-Smad2/3,changes in liver bile acid receptors and transporters FXR,TGR5,NTCP,and MRP2 in Tibetan Cold-Liver model.Clarify effect of JGST on liver Injury in Tibetan Cold-Liver Model.By observing the effect of JGST on the intestinal damage,the intestinal inflammation-related factors IL-6,IL-10 in transcription levels in Tibetan Cold-Liver model.Clarify the effect of JGST on intestinal injury of Tibetan Cold-Liver model.By observing the changes of 25 main bile acid components in the serum,the effect of JGST on the serum bile acid of Tibetan Cold-Liver model was clarified.Results1.Difference between acute cholestasis model and chronic cholestasis model in gut-liver axis(1)Difference of liver injury between acute cholestasis model and chronic cholestasis modelSerum ALT,AST,AKP,TBA,liver TGFβ1,IL-1β,TIMP1,MMP2,and MMP13 were increased in both acute and chronic cholestasis models.αsma decreased in acute cholestasis models,and significantly increased in chronic cholestasis models.The chronic cholestasis model showed more severe bile duct hyperplasia.The liver Masson staining showed that the chronic cholestasis model had more severe collagen deposition.It is suggested that the chronic cholestasis model has more obvious fibrotic damage.(2)Difference of intestinal injury between acute cholestasis model and chronic cholestasis modelZO-1 and Occludin expression decreased in both acute and chronic cholestasis models,intestinal IL-10 increased in both models,IL-1β and IL-6 significantly increased in chronic cholestasis model group,MDA increased in chronic cholestasis model group,SOD declined in chronic cholestasis model group,and GSH and GSH/GSSG decreased in the chronic model group.It is suggested that the chronic cholestasis model has more severe inflammatory and oxidative damage.(3)Changes in fecal bile acid in acute and chronic cholestasis modelsFecal bile acids were generally reduced in the acute cholestasis model group,while toxic bile acids were elevated in the feces of the chronic cholestasis model group.It is suggested that in the chronic cholestasis model,intestinal toxic bile acid increase may be the cause of intestinal damage.2.Regulation of JGST on gut-liver axis injury in chronic cholestasis model(1)Effect of JGST on liver injury in chronic cholestasis modelJGST can improve liver pathological staining,reduce serum ALT,AST,AKP,TBA,down-regulate liver TGFβ1,reduce collagen deposition,reduce P-Smad2/3 in nuclear,reduce liver asma,TIMP1,MMP2,MMP13,and increase liver FXR,MRP2,NTCP expression.It is suggested that JGST can reduce liver damage in chronic cholestasis models.(2)Effect of JGST on intestinal injury in chronic cholestasis modelJGST can improve intestinal pathological damage,increase intestinal ZO-1,Occludin expression;up-regulate intestinal GSH,NQ01,HO-1,GSH/GSSG;down-regulate GSSG;increase the transcription of IL-10,IFNγ,and reduce IL-6,IL-1β;Increase intestinal FXR transcription.It is suggested that JGST can improve the intestinal oxidation and inflammation damage in chronic cholestasis model.3.Differences of gut-liver axis injury between Tibetan Cold-Liver model and chronic cholestasis model(1)Liver damage caused by Tibetan Cold-Liver modelIn the Tibetan Cold-Liver model,serum ALT,AST,AKP,TBA increased,liver TGFβ1 increased,liver TIMP1,MMP2,MMP13 increased,but there was no significant difference compared with chronic cholestasis model,and liver bile duct hyperplasia in Tibetan Cold-Liver model was more Severe,P-Smad2/3 increased in nucleus and asma increased significantly.It is suggested that liver fibrosis is more severe in Tibetan Cold-Liver model.(2)Intestinal damage of Tibetan Cold-Liver modelIn the intestine of Tibetan Cold-Liver model,intestinal villi were broken,ZO-1 expression decreased,IL-10 decreased,IL-1β increased,GSH decreased,MDA increased,and there was no significant difference with chronic cholestasis model.Compared with the chronic cholestasis model,GSSG increased significantly,GSH/GSSG decreased,and Occludin expression decreased.It is suggested that in the chronic cholestasis model,the oxidative damage of the intestine is exacerbated.(3)Changes in serum bile acid of Tibetan Cold-Liver modelCompared with the chronic cholestasis model,the Tibetan Cold-Liver model has more toxic bile acid in serum.It suggested that bile acid liver-gut circulation disorder in Tibetan Cold-Liver model.4.Effect of JGST in gut-liver axis injury in Tibetan Cold-Liver model(1)Effect of JGST on liver damage in Tibetan Cold-Liver modelJGST can reduce liver bile duct hyperplasia,reduce serum ALT,AST,AKP,TBA,down-regulate liver TGFβ1,reduce collagen deposition,reduce P-Smad2/3 nuclear,reduce liver asma,TIMP1,MMP2,MMP13,and increase liver FXR,MRP2,NTCP expression.It is suggested that JGST can reduce liver damage in Tibetan Cold-Liver model.(2)Effect of JGST on intestinal injury in Tibetan Cold-Liver modelJGST can reduce intestinal pathological damage,increase intestinal ZO-1,Occludin expression,up-regulate intestinal GSH,NQO1,HO-1,GSH/GSSG,and down-regulate GSSG;increase IL-10 transcription and decrease IL-1β transcription;Increase intestinal FXR transcription.It is suggested that JGST can improve intestinal oxidation and inflammation damage in Tibetan Cold-Liver model.(3)Effect of JGST on serum bile acid profile of Tibetan Cold-Liver modelJGST can reduce the content of toxic bile acid in the serum of Tibetan Cold-Liver model.It is suggested that JGST can regulate the serum bile acid of Tibetan Cold-Liver model.Conclusion1.Acute cholestasis model and chronic cholestasis model are both suffered in gut-liver axis damage.Chronic cholestasis model has more severe g,which is mainly reflected in intestinal oxidative inflammation damage and liver fibrosis.2.JGST can reduce the gut-liver axis damage in chronic cholestasis model.3.Compared with the chronic cholestasis model,the Tibetan Cold-Liver model has more severe gut-liver axis damage,which is manifested by liver fibrosis and intestinal oxidative.4.JGST can reduce the gut-liver axis damage of Tibetan Cold-Liver model. |
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