The Mouse Model And Clinical Cohort Research About Acute On Chronic Liver Failure

Background and aims: Acute on chronic liver failure(ACLF)is a clinical syndrome occurred among chronic liver disease or cirrhosis patients.Establishing a mouse model of chronic liver disease is the basic of developing the ACLF mouse model.This part is to create a kind of alcoholic related cirrhosis or hepatitis mouse model before the ACLF course.Methods: Liber-De Carli liquid alcohol diet feeding with weekly ethanol binge for 12 weeks can establish a chronic alcoholic liver disease mouse model.During the 12 weeks period,the behavior of the mouse were closely observed,and the change of the ALT,AST and some inflammatory factors were tested.The HE staining,Sirius red staining,oil red staining were made in the liver tissue to observe the pathological changes in this model,while the immunohistochemistry of MPO,CYP2E1 staining and spleen Treg cells frequency were detected to observe the mechanism under this model.Results: The mouse of ALD group in the modeling period undergo a reduction of appetite and malnutrition,following by less activities,unhairing,and loss of gloss of the fur.In the 8th week,some mouse appeared manic,irritability mental performance.In the 4th week,the oil red O staining of the liver tissue in ALD group showed a wide range of steatosis,and mild fibrosis can be seen in the sirus red staining in 8th week,at the same time,HE staining showed inflammatory cells infiltration.At the end of modeling period,it can be observed in more than half of the model group mouse that the liver appeared obvious fiber deposition and inflammatory cells infiltrated with increased serum AST and ALT level,myeloid cells infiltrating in the images of the intrahepatic MPO immunohistochemical.And the CYP2E1 immunohistochemical staining in ALD group had a showed significantly liver injury caused by ethanol.In addition,the spleen lymphocytes flow detection showed that the frequency of Treg cells declined in the disease model group compared with the control group.Conclusion: The 12-weeks Liber De Carli alcoholic liver disease mouse model combined with weekly ethanol gavage cam mimic the natural development of the chronic alcoholic liver disease.And the simple and flexible methods can result to ideal pathologic change.Background and aims: The alcoholic ACLF is a common type of ACLF in the world,but it still lacks a proper mouse model that fully emulates the natural course of ACLF to investigate the underlying mechanism in ACLF.This part aims to develop a mouse model on the basis of the ALD mouse model in the former part.Methods: Based on the former alcoholic liver disease mouse model,the E.coli was injected intraperitoneal to induce acute bacterial peritonitis to complete the acute change in ACLF.The serum AST,ALT,IL-6,IL-10,TNFa and MCP-1 were detected in both the ACLF group and control group with same infection.The HE staining,the liver tissue MPO,CYP2E1 immunohistochemically staining were used to observe the pathological changes after infection.In addition,TNFa receptor blocker was intraperitoneal injected 16 hours before infection to observe the change in survival rate of the ACLF model group.Results: After E.coli infection,the serum AST,ALT of ACLF group was significantly increased compared with the control infection group and the proinflammatory cytokines like IL-6 and TNFa,MCP-1 were also increased obviously.The neutrophil and monocyte/macrophage cells gathered in liver tissue.In addition,hydronephrosis were observed in some ACLF group mouse(2/8).With pre-injection of the TNFa receptor blocker,the survival rate of ACLF improved significantly.Conclusion: Intraperitoneal E.coli injection in the ALD mouse can simulate clinical ACLF natural progression,establishing an ideal alcoholic acute liver failure animal model.The TNFa plays an important role in the deterioration in this ACLF model.Background and aims: RNA sequencing and analysis has become an important technique for the study of pathophysiological mechanisms and the discovery of key molecules.On the basis of the ACLF mouse model,the total RNA of ACLF mouse liver during modeling was sequenced and analyzed,especially the changes of TNFa related pathway.Methods: Normal group,alcohol feeding-2w group,alcohol feeding 12 w group,infected ALD mouse,infected normal mouse were respectively extracted of the total RNA from the liver tissue and the RNA-sequencing was done among the samples.The differential genes of TNF signal pathway and toll-like receptor pathway were analyzed in the transcriptional sequencing data.Results: The toll-like receptor pathway changed slightly in the early stage of chronic alcoholic feeding in mice,and it was not obviously changed during the whole chronic stage.After acute infection,the TLR signaling pathway was significantly activated,and TNFa,IL6,IL1,and CD40 were increased of N1 h and E1 h.In the 2weeks alcohol feeding mice,TNF signaling pathway mainly showed a small number of gene reductions in apoptosis function,such as CREB,CCL2 and IL1.At 12 weeks,the difference gene in this pathway was still in a minority,but it was changed from downregulation to up-regulated,and the up-regulated genes were TNFR1,MKK3,CXCL10 and CCL5.In the Acute phase phase,the pathway of mice in infected groups were activated,CCL2,CXCL10 and IL1 b,Fos protein significantly developed and affected the systemic inflammatory response,leukocyte activation,cell survival.But on the whole TNF pathways downstream expression factor significantly increased at the same time,the normal mice infected also show a negative balance adjustment ability,RIPK3,MLKL,Drp1 in the necroptosis pathways lowered obviously,casp2/3 in apoptosis negatively regulated and some of the classic factors in MAPK pathway of also express less.Conclusion: The balance adjusted function of negative regulation of TNF signaling pathway have been impaired and the silence function of necroptosis was faded away.Background and aims: There are several studies shown that previous decompensation history is one of the important factors affecting ACLF occurrence and deterioration among cirrhosis patients.This history also effects the clinical characteristics and severity of ACLF patients.This section intends to analyze the influence of previous decompensation history in HBV-related cirrhosis patients and HBV-related ACLF patients in the clinical features and prognosis.Methods: 890 cases were enrolled into this cohort from the hospitalized patients from 2005 to 2010 in Ren Ji hospital.All enrolled patients were hospitalized due to acute liver decompensation and the main etiology of their cirrhosis was constantly hepatitis B infection.The cohort was divided into FD(with first decompensation)group and PD(with previous decompensation history)group according to previous liver decompensation history.The statistics were made to analyze the differences about clinical manifestations and outcomes between the two groups,and further analysis was to observe the similarities and differences between the ACLF patients of the two groups.Results: The AST,ALT,TB,the WBC in the FD patients in were significantly higher than PD group,and the incidence of ACLF in FD group was also higher than PD group(39.2% vs 29.2%,p = 0.002),However,the ACLF grades between the two groups have no significant statistical difference.In addition,there was no statistically difference in both short-term(28-days)and long-term(1year)mortality between the PD ACLF patients and FD ACLF patients.Conclusion: There is a higher probability of ACLF occurrence after acute decompensation in the patients with compensated cirrhosis,but the history of previous decompensation has no significant effects in the prognosis of ACLF patients.