Expression Of IL-6/STAT3/IL-17A In Peripheral Blood Of Patients With CHB And Its Associated Acute On Chronic Liver Failure And Its Relationship With Th17/Treg Imbalance

Objective(s):Study of signal transduction and activators of transcription 3(STAT3)mRNA,interleukin-17A(IL-17A)and interleukin-6(IL-6)expression in the peripheral blood of chronic hepatitis B(CHB)and hepatitis B virus(HBV)-associated acute on chronic liver failure(ACLF)and its relationship with Th17/Treg imbalance.Methods:Sixty patients,including 30 patients with CHB and 30 patients with HBV-associated ACLF,admitted to the Department of Geriatric Gastroenterology of the First Affiliated Hospital of Kunming Medical University were included in this study,18 contemporaneous healthy physical examination cases were also included as normal controls.The frequency of Th17 cells and Treg cells were detected by flow cytometry(FCM).The expression levels of peripheral blood STAT3 mRNA,retinoid related orphan nuclear receptor gamma t(RORγt)mRNA and forkhead/winged helix transcription factor 3(FOXP3)mRNA were determined by real-time quantitative reverse transcription polymerase chain reaction(q RT-PCR).The expression levels of IL-17A and IL-6 in serum were detected by enzyme linked immunosorbent assay ELISA).Results:1.The ratios of Th17/CD4~+T cells and Th17/Treg in chronic hepatitis B group were higher than those in normal group(P<0.05).Th17/CD4~+T cells in peripheral blood of patients with HBV-related ACLF were higher than those in the normal group(P<0.05),but there was no significant difference in the three indexes between the chronic hepatitis B group and the HBV-related ACLF group(P>0.05).There was no significant difference in Treg/CD4~+T cells among the three groups(P>0.05).The expression levels of RORγt mRNA and FOXP3 mRNA in peripheral blood of patients in HBV-related ACLF group were higher than those in normal group(P<0.05).Th17/CD4~+T cells in peripheral blood of patients with chronic hepatitis B were positively correlated with HBV DNA load(r_s=0.770,P<0.001).Th17/CD4~+T cells in peripheral blood of patients with HBV-related ACLF were positively correlated with HBV DNA load(r_s=0.632,P<0.001).2.The expression levels of STAT3 mRNA in peripheral blood and serum IL-17A in chronic hepatitis B group were higher than those in normal group(all P<0.05).The expression levels of STAT3 mRNA in peripheral blood,serum IL-17A and IL-6 in HBV-related ACLF group were higher than those in normal group(all P<0.05).There was no significant difference in the expression of STAT3 mRNA,serum IL-17A and IL-6 in peripheral blood between chronic hepatitis B group and HBV-related ACLF group(all P>0.05).The expression levels of STAT3 mRNA,IL-17A and IL-6 in peripheral blood of patients with chronic hepatitis B were positively correlated when compared to each other(r_s=0.715,P<0.001;r_s=0.536,P=0.002;r_s=0.456,P=0.011).The expression levels of STAT3 mRNA,IL-17A and IL-6 in peripheral blood of patients with HBV-related ACLF were positively correlated when compared to each other(r_s=0.476,P=0.008;r_s=0.644,P<0.001;r_s=0.644,P<0.001).The expression levels of STAT3 mRNA in peripheral blood,serum IL-17 and IL-6 in patients with chronic hepatitis B were positively correlated with HBV DNA load(r_s=0.456,P=0.011;r_s=0.565,P=0.001;r_s=0.411,P=0.024).The levels of STAT3 mRNA and IL-17 in peripheral blood of patients with HBV-related ACLF were significantly correlated with serum HBV DNA level(r_s=0.429,P=0.018;r_s=0.403,P=0.027).3.Th17/CD4~+T cells in peripheral blood of patients with chronic hepatitis B were positively correlated with serum IL-6,IL-17A and STAT3 mRNA expression levels(r_s=0.483,P=0.007;r_s=0.628,P<0.001;r_s=0.483,P<0.001).The ratio of Th17/Treg was negatively correlated with FOXP3 mRNA(r_s=-0.411,P=0.024).Treg/CD4~+T cells were positively correlated with FOXP3 mRNA(r_s=0.409,P=0.025).Th17/CD4~+T cells in peripheral blood of HBV-related ACLF patients were positively correlated with serum IL-6,IL-17A,STAT3 mRNA,RORγt mRNA and FOXP3 mRNA expression levels(r_s=0.510,P=0.004;r_s=0.526,P=0.003;r_s=0.673,P<0.001;r_s=0.455,P=0.012;r_s=0.461,P=0.001).Treg/CD4~+T cells were positively correlated with IL-17,RORγt mRNA and FOXP3 mRNA(r_s=0.381,P=0.038;r_s=0.397,P=0.030;r_s=0.409,P=0.025).Th17/Treg ratio was positively correlated with STAT3 mRNA(r_s=0.393,P=0.032).Conclusion(s):1.There is an imbalance of Th17/Treg in peripheral blood of patients with chronic hepatitis B and HBV-related ACLF,and the increase of Th17 cells is dominant.At the same time,Th17/CD4~+T cells was positively correlated with HBV viral load.2.Abnormally high expression of STAT3 mRNA,IL-17A and IL-6 were present in the peripheral blood of patients in both the chronic hepatitis B and HBV-associated ACLF groups,while both were positively correlated with each other and their expression levels were positively correlated with HBV viral load.3.The IL-6/STAT3/IL-17A positive feedback loop is related to the imbalance of Th17/Treg,which may affect each other and indirectly affect the occurrence and development of the disease.