Expression And Diagnostic Value Of Plasma MicroRNA-34a In Liver Fibrosis Of Different Degrees Of Hepatitis B

Background and objectiveChronic hepatitis B(CHB)is caused by long-term hepatitis B virus(HBV)infection,which can progress to liver fibrosis,liver cirrhosis,and eventually lead to end-stage liver disease or hepatocellular carcinoma(HCC).Liver fibrosis is an important intermediate process in the development of liver disease in patients with CHB.The diagnosis of CHB patients with different degrees of liver fibrosis,monitoring the progress of liver fibrosis and appropriate treatment can make the progress of the disease slow down or stable in a certain stage,or even tend to improved.Liver biopsy is not easily accepted by patients due to its invasive nature and possible complications.Instantaneous elasticity imaging of the liver(FibroScan~?)is a relatively mature noninvasive technique and detecting liver hardness value to evaluate the degree of liver fibrosis,which has a correlation with the staging of pathological diagnosis and has been included in the guide.The aspartate aminotransferase-to-platelet ratio index(APRI)and the fibrosis index based on four factors(FIB-4,including age,alanine aminotransferase(ALT),aspartate aminotransferase(AST)and platelet count)can also be used to evaluate the degree of liver fibrosis,but ALT and AST change dramatically in the acute exacerbation of CHB,which may affect the accuracy of liver fibrosis staging.MicroRNAs(miRNAs)are a class of~22 nucleotide long,single-stranded non-coding small RNAs,which mainly play a role in post-transcriptional regulation.The miR-34 family contains three microRNAs encoded by two different genes:miR-34a,miR-34b and miR-34c.Previous studies have confirmed that miR-34 family is a direct target of tumor suppressor gene p53,and its ectopic expression can induce apoptosis and cell-cycle arrest,suggesting that miR-34 family may play an important inhibitory role in tumor and other related diseases.MiR-34a is considered as a tumor suppressor molecule for various tumors,which is differentially expressed and plays an inhibitory role in HBV infection,HBV-associated HCC and other diseases,but the difference in the expression in patients with different degrees of CHB liver fibrosis remains to be explored.MicroRNA is not easy to degrade in blood and has high detection repeatability and specificity,which can be used as a sensitive indicator for monitoring disease progression at the molecular level.In this study,the differences in the expression levels of plasma miR-34a in CHB patients with different liver fibrosis degrees and healthy controls were detected to investigate the monitoring effect of miR-34a on the condition of different liver fibrosis degrees.And a comparative analysis was made with APRI and FIB-4 to explore a better monitoring index on the molecular level or to conduct combined monitoring.Materials and methods1.Patients and controls:Plasma samples from 105 patients with CHB were enrolled in this study from the Department of Infection of the First Affiliated Hospital of Zhengzhou University as experimental group.According to the FibroScan~?detection of liver hardness values,105 patients included 35 patients with mild liver fibrosis(F0F1 group),35 patients with moderate liver fibrosis(F2F3 group)and 35patients with severe liver fibrosis and early liver cirrhosis(F4 group).Plasma samples from 35 healthy volunteers were collected as control group.2.Detection of plasma miR-34a:The relative expression level of miR-34a in plasma samples of the experimental group and control group was detected by quantitative real time polymerase chain reaction(qRT-PCR).3.The APRI and FIB-4 were calculated according to the clinical data of the patients and controls.4.Statistical analysis:The SPSS 17.0 and Prism 5.0 statistical software were used for data comparative analysis and graph drawing.The receiver operating characteristic curve(ROC)and area under curve(AUC)were used to analyze the diagnostic value of miR-34a,APRI and FIB-4 on different degrees of liver fibrosis in patients with CHB.Results1.The relative expression level of plasma miR-34a in patients with CHB liver fibrosis was significantly lower than that in the control group(P<0.001).In CHB patients with different degrees of liver fibrosis,the relative expression level of plasma miR-34a in F4 group was significantly lower than that in F0F1 group(P=0.003).2.The level of APRI in patients with CHB liver fibrosis was significantly higher than that in the control group(P<0.001).In CHB patients with different degrees of liver fibrosis,F4 group had a higher APRI level than F0F1 group(P=0.003).3.The level of FIB-4 in patients with CHB liver fibrosis was significantly higher than that in the control group(P<0.001).In patients with CHB,the level of FIB-4 in F2F3 group was higher than that in F0F1 group(P=0.017)and the level of FIB-4 in F4 group was significantly higher than that in F0F1 group(P<0.001).4.ROC results showed that the AUCs of plasma miR-34a,APRI and FIB-4 in the diagnosis of CHB were 0.726(0.619-0.833),0.978(0.959-0.997)and 0.896(0.845-0.948),respectively.The AUCs of plasma miR-34a,APRI and FIB-4 in the diagnosis of moderate and severe liver fibrosis and early liver cirrhosis were 0.699(0.600-0.797),0.830(0.740-0.920)and 0.808(0.724-0.893),respectively.The AUCs of plasma miR-34a,APRI and FIB-4 in the diagnosis of severe liver fibrosis and early liver cirrhosis were 0.704(0.602-0.806),0.757(0.661-0.853)and 0.769(0.672-0.866),respectively.5.As for the combined diagnostic value of patients with CHB,the AUCs of miR-34a+APRI,miR-34a+FIB-4 and miR-34a+APRI+FIB-4 were higher than that of the APRI or FIB-4 separately(0.982 vs 0.978,0.927 vs 0.896,0.983 vs 0.978/0.896).The AUC of APRI+FIB-4 did not increase compared with that of APRI alone(0.978vs 0.978).As for the combined diagnostic value of patients with moderate and severe liver fibrosis and early liver cirrhosis,the AUCs of miR-34a+FIB-4 and miR-34a+APRI+FIB-4 were higher than that of the FIB-4 or APRI separately(0.840vs 0.808,0.850 vs 0.830/0.808),while the AUC of APRI+FIB-4 was lower than that of APRI alone(0.815 vs 0.830).As for the combined diagnostic value of patients with severe liver fibrosis and early liver cirrhosis,the AUCs of miR-34a+APRI,miR-34a+FIB-4 and miR-34a+APRI+FIB-4 increased(0.771 vs 0.757,0.797 vs0.769,0.804 vs 0.757/0.769),while the AUC of APRI+FIB-4 decreased(0.761 vs0.769).Conclusions1.The relative expression level of plasma miR-34a was significantly decreased in patients with CHB liver fibrosis,and its decreased expression was related to the disease progression of liver fibrosis.Plasma miR-34a can be used as a new candidate marker to identify different stages of liver fibrosis in patients with CHB.2.APRI and FIB-4 were obviously increased in CHB patients with different degrees of liver fibrosis.With the aggravation of liver fibrosis,APRI and FIB-4 were increased gradually.3.Plasma miR-34a has diagnostic ability for different stages of liver fibrosis in CHB patients.The panel of plasma miR-34a,APRI and FIB-4 perfects the diagnostic ability in CHB liver fibrosis.