Research On The Mechanism Of Colon-targeted Drug Huchangqingdu Pellets In Blocking Intestinal Endotoxemia Of Acute On Chronic Liver Failure

Objective:To investigate the possible mechanism of Huchangqingdu pellets in eliminating intestinal endotoxemia of acute on chronic liver failure through clinical and animal experimental study,thereby providing theoretical basis for the clinical application of Huchangqingdu pellets in eliminating intestinal endotoxemia of acute on chronic liver failure,and laying the foundation for further research.Methods:1.Clinical research:From January 2013 to October 2016,60 patients in accordan ce with the inclusion criteria of HBV related acute on chronic liver failure with int estinal endotoxemiawere divided into oral lactulose group and oral Huchangqingdupe llets group according to the randomized double-blind,double simulation comparison principle,30 cases for each group.TCM syndrome integral,plasma endotoxin(ET),t umor necrosis factor(TNF-a),interleukin-1(IL-1),interleukin-6(IL-6),interleukin-8(IL-8),interleukin-10(IL-10),blood D-lactic acid(D-LAC),blood monoamine oxidase(DAO),thrombin,urine lactulose/mannitol(L/M)and other changes were observed before a nd after treatment.Compare the different curative effects of two drugs.2.Experimental study:Wistar rats were divided into normal group,model group,Huchangqingdupellets groups(including low,medium and high dose groups),lactulose group randomly,which treated respectively.Observe the general situation andliver function changes of wistar rats.Test the level of TNF-??IL-1?IL-2?IL-6?IL-8 in serum of rats through ELISA assay,detect rat liver tissue CD 14,CD 163 expression and TLR4,NF-?Bp-65expression through immunohistochemical method and Western Blot respectively,and detect the expression of CD14?CD163?TGF-?1?TLR4?NF-?BmRNA through RT-PCR.Results:1.Clinical study:After treatment,the improvement of symptom index and com prehensive curative effect in Huchangqingdupellets group were better than those bef ore treatment(P<0.05).The improvements in aspects of TCM syndrome integral,tot al Bilirubin(TBIL),pre-albumin(PA),prothrombin activity(PTA),plasma endotoxin(ET),inflammatory cytokines,D-LAC,DAO inHuchangqingdu pellets group were mo re obvious than those in lactulose group.(P<0.05).2.Experimental study:Middle and high dose Huchangqingdu pelletscould significantly improve the general condition and liver function indexes of rats,and decrease the levels of TNF-?,IL-1,IL-2,IL-6 and IL-8,up-regulate the expression of CD163 at protein and gene level,and down-regulate the expression of CD14,TLR4 andNF-KB mRNA.In the low dose Huchangqingdu pelletsgroup,the intervention effect on model rats was not obvious.The middle dose Huchangqingdu pellets group was better than lactulose group in improving the body weight,biochemical indexes,reducing inflammatory factors,up-regulating CD 163 and down-regulatingexpression of CD 14,TLR4 andNF-?B(P<0.05).Conclusions:1.Colon-targeted release of huchangqingdu pellets can reduce mortality of acute-on-chronic liver failure(ACLF),improve the clinical syndrome and biochemical indexes,and prevent the release of inflammatory cytokines,improve the permeability of intestinal mucosa and stabilize the internal environment,so as to effectively block the occurrence of intestinal endotoxemia and repair liver injury.The overall effect is better than lactulose.2.Huchangqingdu pellets can regulate the LPS-TLR4 pathway,down-regulate CD 14,TLR4,NF-?BmRN A and up-regulate scavenger receptorCD 163 eliminate inflammatory factor infiltration,reduce liver cell necrosis and apoptosis.It also can protect the intestinal mucosa microvilli and epithelial cell basement membrane,improve cellular tight junction andthus effectively repairing intestinal mucosal injury and mucosal permeability,blocking the progress of IETM and promoting liver structureand functional recovery.This could be the potential mechanism of eliminating acute on chronic liver failure intestinal endotoxemia.