乙型肝炎肝硬化患者外周血IL-35和IL-17的表达及意义

目的观察慢性HBV感染者外周血IL-35和IL-17的表达和临床指标的相关性。方法采集30例慢性乙型肝炎（CHB）患者、79例乙型肝炎肝硬化（LC）患者、14例慢性乙型重型肝炎（CSHB）患者、20例健康对照者的外周血,ELISA法测定外周血IL-35、IL-6、IL-17的浓度,分离制备外周血单个核细胞（PBMC）,实时定量PCR检测PBMC中EBI3、FOXP3、IL-17 mRNA的表达,Western印迹观察PBMC中EBI3、FOXP3、IL-17蛋白表达情况。所有病例及健康对照均进行肝功能检测。结果 LC、CHB和CSHB患者中EBI3、IL-17、FOXP3 mRNA表达水平均显著高于健康对照组,差异有统计学意义（P〈0.01或0.05）;LC组Child-Pugh不同分级患者之间EBI3、IL-17、FOXP3 mRNA表达水平差异明显,EBI3、FOXP3 mRNA随着肝硬化Child-Pugh分级的加重而表达下调;IL-17 mRNA随着肝硬化Child-Pugh分级的加重而上调;蛋白表达和血清浓度的变化趋势与mRNA一致。血清IL-17浓度水平与Alb呈负相关,IL-17浓度水平与TBil、ALT和Child-Pugh分级呈正相关,差异有统计学意义（P〈0.01）,IL-35浓度水平与Alb、Child-Pugh分级负相关,与ALT、TBil无相关性。结论 IL-35和IL-17可能是影响乙型肝炎肝硬化发病的重要因素之一。

Expression and significance of interleukin-35 and interleukin-17 in the blood of patients with hepatitis B related liver cirrhosis

Objective To observe the expression levels and significance of interleukin（IL）-35 and IL-17 in the blood of patients with hepatitis B related liver cirrhosis.Methods The levels of IL-35 and IL-17,which was from peripheral blood of 30 cases with chronic hepatitis B（CHB）,79 cases with liver cirrhosis（LC）,14 cases with chronic severe hepatitis B（CSHB）and 20 normal controls（NC）,were detected by enzyme-linked immunosorbent assay（ELISA）.Real-time polymerase chain reaction（RT-PCR）was used to evaluate the expression levels of EBI3,FOXP3 and IL-17 mRNA in peripheral blood mononuclear cells（PBMC）,while western blot was used to determine protein expression.Liver function of patients and normal controls was measured.Results The expression of EBI3,IL-17 and FOXP3 mRNA in PBMCs from patients with LC,CHB and CSHB was higher than that from normal controls（P0.01 or 0.05）.Significant difference in the level of IL-17 mRNA can be identified among different Child-pPugh classification patients.The levels of EBI3 and FOXP3 mRNA were down-regulated with Child-Pugh grades increased.The level of IL-17 mRNA was up-regulated with Child-Pugh grades increased.Changes of protein expression and concentrations in peripheral blood were consistent with mRNA expression.Serum IL-17 levels were positively correlated with TBil,ALT and Child-Pugh grades,and negatively correlated with Alb.The differences were statistically significant.Serum IL-35 levels were negatively correlated with Alb and Child-Pugh grades while ALT and TBil showed no correlations.Conclusion IL-35 and IL-17 may be critically involved in the pathogenesis of hepatitis B related liver cirrhosis.