Clinical Features And Prognostic Evaluation Of Patients With Drug-induced Liver Injury And The Effect Of Endoplasmic Reticulum Stress On Rifampicin And Isoniazide-induced Liver Injury In Rats

Objective(1)To summarize the clinical characteristics of patients with durg-induced liver injury(DILI)and investigate the prognostic evaluation factors of DILI.(2)To explore the role of endoplasmic reticulum stress(ERS)in rifampicin(RFP)and isoniazid(INH)induced liver injury in rats and its possible mechanisms.Methods(1)The clinical data of hospitalized DILI patients in recent five years were analyzed retrospectively,factors effecting on prognosis were selected by using Logistic regression analysis.(2)32 male Sprague-Dawley(SD)rats were randomly divided into four groups,each group included eight rats.Control group(N group)rats were given equivalent saline by empty stomach gavage administration on time once daily;RFP group(R group)rats were given RFP 50 mg/(kg*d)once daily by empty stomach gavage administration on time;Model group(M group)rats were given RFP 50 mg/(kg*d)and INH 50 mg/(kg*d)once daily by empty stomach gavage administration on time;PBA group rats were given 4-phenylbutyric acid(PBA)160 mg/(kg*d)once daily by intraperitoneal injection and the durg administration method of INH and RFP were the same to the M group.The rats were sacrificed after 14 days.The blood and liver tissue of rats were collected.Alkaline phosphatase(ALP),total bilirubin(TBIL),direct bilirubin(DBIL),alanine aminotransferase(ALT),aspertate aminotransferase(AST)and total bile acid(TBA)level in the serum of rats were tested by fully automatic biochemical analyzer.The liver pathology change of rats were observed under light microscope through hematoxylin eosin staining.The expression changes of glucose regulated protein78(GRP78),P65,inhibitor kappa b(IκB)and bile salt export pump(Bsep)in rats liver tissue were detected by western blot method.The change of expression and localization of Bsep in hepatocyte membrane was detected by immunohistochemistry.Results(1)Of the 106 patients,female was 61(57.5%),the mean age was 52 years old,Chinese herbal medicines,antibacterial and antipyretic analgesic drugs accounted for the major part of medications.The percentage of hepatocellular pattern was 81.1%,the most common severity grade of DILI was moderate(54.7%),91.5% patients were good prognosis.There was significant difference between Hy ’s law coincidence rate in different severity of DILI(P < 0.05),the incidence rate of liver failure in accordance with Hy’s law in hepatocellular pattern of DILI was 22.9%(11/48).The indicators that affected the prognosis were ALT(OR=1.672,P=0.026)and international normalized ratio(INR)(OR=8.852,P=0.015)were found by multi-factor Logistic regression analysis.(2)Compared with the N group,TBIL,DBIL,ALP and TBA in the serum of rats of R and M group were significantly increased(P<0.05),while the elevation of ALT and AST was not obvious(P > 0.05);Hepatocellular fatty degeneration was observed in R group rats,while the degree of fatty degeneration was increased in rat livers in M group with inflammatory cell infiltration and spotted and focal necrosis;the expression of GRP78 and P65 in rat livers was increased obviously(P < 0.05),meanwhile,IκB and Bsep was significantly decreased(P < 0.05)in R and M group.The average integral optical density(IOD)of Bsep on hepatocyte membrane in R and M group were 3.56± 1.18 and 2.35±1.18,which were significantly decreased than N group(9.63±1.59)(P<0.05),and Bsep on hepatocyte membrane in M group was irregularly distributed.Compared with M group,TBIL,DBIL,PBA and ALP reduced significantly(P < 0.05)in the serum of PBA group rats;liver pathology was also improved obviously;the expression of GRP78 and P65 was obviously down-regulated,IκB and Bsep was significantly up-regulated(P < 0.05).The average IOD of Bsep on hepatocyte membrane was 4.66±1.75,which was significantly increased than M group(P < 0.05),and the distribution of Bsep on hepatocyte membrane in PBA group was more regularer than M group.Conclusion(1)The majority of patients with DILI had a good prognosis.ALT and INR at DILI recognition were the independent factors for predicting the prognosis of DILI.(2)RFP induced chelostasis and hepatocellular fatty degeneration in rats,increased the expression of GRP78,P65 and decreased Bsep,IκB in rat livers and made the down-regulated expression and localization change of Bsep on hepatocyte membrane.(3)PBA decreased the expression of GRP78,P65 and increased Bsep,IκB in isoniazid-rifampin-induced liver injury in rats,and increased the expression and correct localization of Bsep on hepatocyte membrane,which indicated that ERS played an important role in isoniazid-rifampin-induced liver injury.The possible mechanism of RFP and INH induced liver injury is by activating the ERS/NF-κ B pathway,down-regulating expression of Bsep and changing its localization.