Effect Of Celecoxib On P53 And MicroRNA-34a/HNF4α Pathway In Non-Alcoholic Steatohepatitis Mice Models

Objective: To explore the impact of celecoxib on miR-34a/HNF4α pathway through p53 and its healing efficacy on non-alcohol steatohepatitis(NASH),offering a new thought on NASH treatment.Methods: 36 male 7 to 8-week-old c57BL/6 mice were obtained then randomly divided into 3 groups after 1-week adaptive feeding.Control group(CON,n=10),NASH+normal saline group(NASH+NS,n=13),NASH+celecoxib group(NASH+CEL,n=13).The NASH mice models were induced by feeding methionine-choline deficient diet(MCD)daily only.CON and NASH+NS groups were treated with normal saline(0.2ml/10g·W/d)by gastric gavage;NASH+CEL group was received gavage of Celecoxib(0.2mg/10g·W/d,Pfizer Pharmaceuticals LLC).After 4 weeks,all mice were sacrificed under anesthesia after a 12 h period fasting.Blood were collected for measurement of serum ALT,TC and TG level;the liver samples were taken and fixed in 10% formalin(>4h)for HE staining with steatosis,activity and fibrosis score;the remaining liver tissues were collected,stored at-80 ℃for western blot analysis,ELISA and qRT-PCR test,to assaying p53,HNF4 a protein level and the expression of mi R-34 a.Experimental data were analyzed by SPSS software version 22.0.Results: 1.NASH models were successfully built.After 4 weeks treatment with celecoxib,the hepatic steatosis,activity and fibrosis in NASH+CEL group were ameliorated,compared with NASH+NS group,the differences were statistically significant(P<0.05).2.The p53 protein level in NASH groups was significantly elevated,compared with CON group(P<0.05).Whereas the p53 protein level in NASH+CEL group shows a tendency of reduction than NASH+NS group,but there was no statistical significance(P>0.05).3.The relative miR-34 a level in NASH groups was sharply raised,to compare with CON group(P<0.05),after fed with celecoxib,the relative mi R-34 a level illustrated a tendency of decreasing,but there was no statistical significance(P>0.05).4.There was no statistical significance in HNF4α between three groups(P>0.05).5.The body weight and liver wet weight show decrease in MCD NASH models than chow diet mice(P<0.05),whereas the liver index implied no difference(P>0.05).The body weight,liver wet weight and liver index indicated no difference after feeding with celecoxib(P>0.05).6.Serum ALT level was sharply increased while serum TC and TG decreased(P<0.05).After feeding with celecoxib,serum ALT level decreased whereas serum TC and TG level slightly increased in NASH+CEL group when compared with NASH+NS group,but there was no statistical significance(P>0.05).Conclusions: 1.Celecoxib can ameliorate several steatosis,inflammation and fibrosis in MCD NASH mice models;2.There are a decressing tendency of the p53 protein level and the relative miR-34 a level in NASH models after fed with celecoxib;3.The fact that celecoxib attenuates NASH histology may not due to the control of mi R-34a/HNF4α pathway through p53.