MicroRNAs Expression In Liver Tissues And Sera Of Patients With HBV-related Acute-on-chronic Liver Failure And Their Clinical Prognostic Roles

AIM Exploring the micro RNAs expression in serums and liver tissues of patients with Acute-on-Chronic Liver Failure(HBV-AC LF), to examine the possibility of micro RNA as a novel, non- invasive HBV-ACLF prognostic biomarker. Meanwhile, we also explored how micro RNA involve in the pathogenesis of HBV-ACLF by regulating the target gene expression. METHODS Microarray analysis was performed to characterize the mi RN As expression profiles in liver tissues from 1 HBV-AC LF patient and 1 matched healthy control. Then, the selected mi RN As were verified in liver tissues(HBV-ACLF=8, Control=8) and further detected in serums(HBV-ACLF=39, Control=20) by using quantitative real-time Polymerase C hain Reaction(q RT-PCR) technology. According to the prognosis in three months, the 39 HBV-ACLF patients were divided into spontaneous recovery group(n=20) and non-spontaneous recovery group(n=19), which including death(n=9) and liver transplantation(n=10).The mi RN As with differential expression between the two groups were selected and explored their correlationship with MELD score, PTA, INR and PT, respectively.Receiver operating characteristic curve(ROC curve) was used to analyze the prediction value of mi RN A in HBV-ACLF prognosis.Then,target gene prediction software s involving mi RWalk,Target Scan and mi Randa were performed to predict target genes of mi RNA. Then target genes expression were detected by q RT-PCR in liver tissues from HBV-ACLF patients(n=11)and matched healthy control(n=6).RESULTS(1) Microarray analysis showed that, compared with the matched healthy control, 89 mi RN As expressions were up-regulated and 99 mi RN As were down-regulated(folds ≥5) in the HBV-AC LF liver tissue.After literature review, we selected 9 differentially expressed mi RNAs(Mi RNAs with up-regulation expression were mi RN A-21 、 mi RNA-130 a 、 mi RNA-143 and mi RNA-200,those with down-regulation were mi RN A-486-5p、mi RNA-192、mi RNA-194、mi RNA-148、 mi RN A-122) for further study.(2) Compared with 8 matched healthy controls, q RT-PCR verified that mi RN A-130 a, mi RNA-486-5p and mi RN A-200 were up-expressed in liver tissues of 8 HBV-ACLF patients(p <0.05). Except mi RNA-486-5p, the expressions of others were consistent with the microarray results. In the 39 HBV-ACLF patients(age 43.24±2.14,male 31) and 20 age and sex- matched normal control(age 42.05±2.29,male 14), q RT-PCR results showed that, 8 mi RN As including mi RN A-21, mi RN A-130 a, mi RNA-486-5p, mi RNA-192, mi RN A-148, mi RNA-143, mi RN A-122,and mi RN A-200 were markedly upregulated in the serums of HBV-AC LF patients(p <0.05).(3)Compared with non-spontaneous recovery group, serum both mi RN A-130 a and mi RN A-486-5p were upregulated in HBV-ACLF recovered group serum(p <0.05). Further statistical analysis in the 39 HBV-ACLF patients, mi RNA-130 a was negative correlation with MELD score(r =-0.39, p = 0.03)、INR(r =-0.38, p = 0.03) and PT(r =-0.41, p = 0.02),but positively correlated with PTA(r = 0.36, p = 0.05). Whereas,mi RN A-486-5p had no correlation with above coagulation parameters and MELD score(p> 0.05). ROC curve analyzed that there were no significant difference of AUC between mi RNA-130a(AUC=0.74,95% CI0.571-0.91, p=0.02)and MELD(AUC=0.86,95% CI 0.720-0.991, p = 0.00)(p= 0.163). These results suggested that mi RNA-130 a had prognositic prediction value in HBV-ACLF RESULTS patients.(4) Mi RWalk,Target Scan and mi Randa target gene prediction softwares analysis predicted that Glycogen synthase kinase(GSK) was the potential target gene of mi RN A-130. Then q RT-PCR verified that GSK m RNA expression had a downward trend(p = 0.078) in the liver tissues from HBV-AC LF patients, compared with the matched healthy controls.CONCLUSIONS(1) Up-regulated mi RNA-130 a in both serum and liver tissues of HBV-ACLF patients and its prediction value in clinical prognosis, suggested that mi RN A-130 a might be a novel potential non-invasive prognostic indicator of HBV-AC LF.(2) In mi RNA regulatory networks, mi RNA-130 a may involve the pathophysiology of HBV-ACLF by downregulating the expression of GSK.