Expression profiles of exosomal microRNA in the different phases of natural history in chronic hepatitis B patients / 中华传染病杂志

ABSTRACT

Objective To explore the expression profiles and their clinical significance of serum exosomal microRNA (miRNA ) in the different phases of natural history in chronic hepatitis B (CHB ) patients .Methods A total of 92 treatment-naive CHB patients in Shanghai Public Health Clinical Center between January 2014 and January 2017 were retrospectively studied .The cases in immune tolerant (IT) phase ,immune reactive (IR) phase ,inactive carrier (IC) phase and HBeAg-negative CHB (ENH) phase were 24 ,24 ,24 ,and 20 ,respectively .Exosomes were isolated by ExoQuick solution from 250μL serum . The expressions of the surface protein markers LAMP2 and TSG101 in serum exosomes were determined by Western blotting . The expressions of miRNA-122 , miRNA-125a , miRNA-29c , miRNA-200c in exosomes were detected by real-time fluorescent quantitative PCR .The Kruskal-Wallis H test and Mann-Whitney U test were used to determine intergroup differences . The correlation coeffcients ( r) were calculated using Spearman′s correlation .Receiver operating characteristic (ROC) and the area under the curve (AUC ) were used to calculate the diagnostic values . Results Western blotting showed that exosome-specific markers LAMP2 and TSG101 were positive .The levels of serum exosomal miRNA-122 , miRNA-125a ,miRNA-29c and miRNA-220c were significantly different in CHB patients in different phases (H=41 .06 ,29 .31 ,49 .14 and 31 .73 ,respectively ,all P<0 .05) .Based on the results of liver function test and liver biopsy , serum exosomal miRNA-122 , miRNA-29c and miRNA-200c in inflammation group were down-regulated (U = 804 ,317 and 574 ,respectively ,all P< 0 .05) ,while miRNA-125a was up-regulated (U=279 ,P<0 .01) compared with non-inflammation group .The level of exosomal miRNA-200c was negatively correlated with ALT (r= -0 .3932 ,P<0 .01) ,while the level of miRNA-125a was positively correlated with ALT (r=0 .5981 , P<0 .01) .AUC of the above exosomal miRNA were 0 .6193 ,0 .8396 ,0 .8243 and 0 .6883 ,respectively .The highest AUC was miRNA-125a with the sensitivity of 84 .62％ and the specificity of 74 .47％ .AUC of ALT discrimination for liver inflammation was 0 .7953 ,with the sensitivity of 81 .08％ and the specificity of 70 .97％ .Conclusions The serum exosomal miRNA levels are significantly different among the different phases of natural history in CHB patients .Inflammation-associated exosomal miRNA is expected to be the non-invasive diagnostic biomarkers of liver inflammation and is of great benefit for determining the best time for clinical medication of CHB patients .