The Study Of Detoxication And Activating Blood Stasis Treatment Effects And Mechanism To Endotoxic Level Of Acute-on Chronic Liver Failure

Objective1.Through clinical observation to evaluate the clinical effects and survival rate of detoxication and activating blood stasis treatment for HBV-ACLF.2.Through the "Eleventh five years" and "Twelfth five years"national science and technology major projects to research detoxication and activating blood stasis treatment effects for endotoxic level of HBV-ACLF,also the effects of endotoxic to HBV-ACLF.3.To explore the mechanism of detoxication and activating blood stasis treats ACLF endotoxmia through P2X7R/AN06 ion channels by ACLF animal model.Methods1.Our study used randomized controlled and single-blind.Enrolled HBV-ACLF patients trail and control group with 2:1.Control group treated with conventional internal western medicine,such as nutritional support,antiviral treatment,protect liver and decline aminopherase,improve the hepatic microcirculation,promoting hepatocyte regeneration,artificial liver if necessary.The trail group treated with the conventional internal western medicine,plus prescription of cool blood,detoxification and remove blood stasis or tonifying Qi,detoxification and remove blood stasis blood after syndrome differentiation of Chinese medicine.Two groups were treated for 8 weeks and 120 weeks follow-up.2.During eight weeks treatment,tested liver function,renal function,coagulation function,electrolyte,blood lipid and endotoxin every week.Tested five quantitative of hepatitis B and HBV DNA,T lymphocyte subsets,cytokines(TNF-?,IFN-?,IL-2;IL-4,IL-5,IL-6,IL-10),hepatic ultrasound etc at certain time.The follow-up of weeks gave conventional laboratory tests and observed the survival rate.3.Tested endotoxic levels of liver failure patients from 20 hospitals at 0 week,4th week,8th week and 12th week by ELISA limulus reagent,then analysis.4.Made ACLF rat model with immune method.Gave prescription of cool blood,detoxification and remove blood stasis to the ACLF model 1 week by intragastric administration,then hocused the rats and taken blood from abdominal aorta and collected liver for test and liver HE staining,immunohistochemistry,Western blot,quantitative PCR to observation P2X7R,AN06 and mDC expression.Results1.?The baseline data of age,sex,the illness and 0 week,the differences had no statistical significant.During 8 weeks treatment,after treatment of 1st week,4th week and 8th week,trail group indexes of TB-L,ALT,AST,CHE,PT,PTA,INR lower compared with control group,the differences had statistical significant(P<0.05 or 0.01).?4th week,8th week and 12th week of the trail group total effective rate were 86.49%,89.19%and 90.91%respectively,the control group was 75%,75%and 83.3%respectively.?At 4th week and 8th week,trail group compared with control group had differences on the number of patients with risk grade of MELD scores and CTP scores,the differences had statistical significant(P<0.05)?Kaplan-Meier survival analysis of trail group survival rates were 90%and 84.6%in 12th week and 72th week,respectively,the control group were 80% and 75%,respectively.At 120th follow-up week,the survival rates of the trail group and the control group were 77.5%and 50%,respectively,the differences had statistical significant(P<0.05).?ROC curve shown that CTP score AUROC=0.755 was higher than the MELD score AUROC= 0.562,the differences had statistical significant(P<0.05),the Cut-off value of CTP was 9.5,the sensitivity and specificity were 78.9% and 65.9%,respectively.?We did not find adverse events related with drug after the treatment.2.?The trail had lower endotoxic level compare with control group at 4th week and 8th week after treatment,the differences had statistical significant(P<0.05).? Results of 11th five years:The trail group endotoxic level was lower than control group at 4th week,8th week and 12th week,whether the early stage ACLF,middle stage ACLF or late stage ACLF,the differences had statistical significant(P<0.01).The survival rate of trail group was higher than control.3.?The number of T lymphocyte subsets had increased in trail group at 4th week and 8th week,which compared with control,the differences had statistical significant(P<0.05).?the level of IL-2,IL-6,TNF-a and IFN-y in Trail group had reduced,but the IL-4and IL-10 level had increased,that compared with control group at 4th week.IL-2,IL-5,IL-10 level had increased and IL-4,IL-6,IFN-ylevel had reduced at 8th week compared with control.These variation had statistical significant(P<0.05 or 0.01).4.Detoxication and activating blood stasis treatment could lower the expression of P2X7R,AN06 and mDC in ACLF animal model liver tissues,the data of RT-PCR relative gray-value showed statistical significant(P<0.05 or 0.01).Conclusion1.?Detoxication and activating blood stasis treatment can improve the liver function and blood coagulation function of early stage HBV-ACLF,and accelerate HBV-ACLF recovery.Detoxication and activating blood stasis treatment is safe and no adverse reactions.?Detoxication and activating blood stasis treatment can improve the prognosis of HBV-ACLF patients,decrease the mortality.2.The death of HBV-ACLF relate with endotoxic level.3.CTP score can predict adverse outcome of early stage HBV-ACLF more accuracy than MELD score,the cut-off value of CTP score is 9.5.4.Detoxication and activating blood stasis treatment can reduce the production of ACLF patients' endotoxic level,prevent and control the endotoxmia,also can protect the hepatoctye and reduce mortality.5.Detoxication and activating blood stasis treatment may reduce the expression of P2X7R and AN06 in liver tissue,reduce the number of DC migrate to liver,thus reduce the inflammation responses and decline endotoxemia.