Objective To investigate the changes of Th1/Th2 cells and related cytokine levels in chronic hepatitis B (CHB) fibrosis.Methods Forty-six patients with CHB fibrosis underwent liver biopsy during March and October,2011.According to the stage of fibrosis,the patients were divided into S0-1 group (n =15),S2-3 group (n =20) and S4 group (n =11).Ten healthy subjects served as controls.The frequencies of circulating Th1,Th2 cells were detected by flow cytometry.The expressions of interferon-γ (IFN-γ) and interleukin 4 (IL-4) mRNA in peripheral blood mononuclear cells (PBMCs) were detected by real-time quantitative PCR.The serum IFN-γand IL-4 concentrations were determined by enzyme-linked immunosorbent assays.Intrahepatic expressions of IFN-γ and IL-4 were detected by immunohistochemical staining.Differences between groups were analyzed using non-parametric Kruskal-Wallis H test,followed by Mann-Whitney U test for multiple comparisons.Logistic regression was used for multivariate analysis.Results With the degree of liver fibrosis exacerbations,the peripheral Th1/Th2 cells frequencies ratio,IFN-γ/IL-4 mRNA ratio in PBMCs,serum IFN-γ/IL-4 ratio and intrahepatic IFN-γ/IL-4 ratio were declined (x2 =36.259,40.822,26.321 and 31.852,respectively,all P ＜ 0.05).Serum and intrahepatic IFNγ/IL-4 ratio were negatively associated with the stage of liver fibrosis (r =-0.616 and-0.531,P ＜0.01).Logistic regression analysis showed that AST,PT and the serum IFNγ/IL-4 ratio were the risk factors for significant liver fibrosis (S2-4) (OR =5.933,95％ CI:1.324-26.586,P =0.02; OR =12.866,95％CI:1.746-94.788,P =0.01; OR=4.755,95％CI:1.034-21.862,P =0.04).Conclusions The CHB patients has imbalanced Th1/Th2 ratio.With the degree of liver fibrosis exacerbations,Th1/Th2 cytokines drift into Th2 lymphocyte sub-cluster,which suggests that Th1/Th2 imbalance may be involved in the pathogenesis of CHB fibrosis.%目的 探讨Th1/Th2细胞及其分泌的细胞因子在慢性乙型肝炎(CHB)肝纤维化患者发生发展中的作用.方法 选取2011年3-10月在河北医科大学第三医院感染科接受肝组织病理活检的CHB肝纤维化患者46例,根据纤维化分期分为S0～1期15例,S2～3期20例,S4期11例三组,另取同期健康志愿者10名作为健康对照组.用流式细胞术和实时荧光定量PCR检测外周血Th1、Th2细胞频率和单个核细胞(PBMCs)中干扰素γ(IFN-γ)、白细胞介素4(IL-4)基因表达；用ELISA法检测血清IFN-γ和IL-4水平；用免疫组织化学染色检测肝组织IFN-γ和IL-4表达.应用Spearman相关分析、Kruskal-Wallis H检验、Logistic逐步回归方法分析Th1/Th2细胞及其细胞因子在CHB肝纤维化患者外周血和肝组织中的变化.结果 不同肝纤维化分期的CHB患者外周血Th1/Th2细胞频率的比值、PBMCs中IFN-γ/IL-4 mRNA、血清中IFN-γ/IL-4及肝内IFN-γ/IL-4差异均有统计学意义,并随肝纤维化程度加重逐渐下降(x2=36.259、40.822、26.321和31.852,P＜0.05).血清及肝组织中IFN-γ/IL-4比值与肝纤维分期均呈负相关(r=-0.616和-0.531,P＜0.01).Logistic回归分析显示:AST、凝血酶原时间(PT)和血清IFN-γ/IL-4比值是发生显著肝纤维化(S2～4)的三个独立危险因素(OR=5.933,95％ CI:1.324～ 26.586,P=0.02; OR=12.866,95％ CI:1.746 ～94.788,P=0.01; OR=4.755,95％ CI:1.034～21.862,P=0.04).结论 CHB患者体内存在Th1/Th2失衡,随着肝纤维化进展,外周血及肝脏内均呈现Th2应答的极化现象,这可能是CHB肝纤维化发生发展的一种机制.
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