慢性乙型肝炎肝纤维化患者Th1／Th2细胞及其细胞因子的变化

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IL4表达。应用Spearman相关分析、Kruskal—WallisH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-4mRNA、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（χ2=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／IL4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。

Changes of Th1/Th2 cells and related cytokine levels in patients with chronic hepatitis B fibrosis

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 （luring March and October, 2011. According to the stage of fibrosis, the patients were divided into SO - 1 group （n = 15）, S2-3 group （n =20） and S4 group （n = 11）. Ten healthy subjects served as controls. The frequencies of circulating Thl, Th2 cells were detected by flow cytometry. The expressions of interferon- γ （IFN-γ） and interleukin 4 （IL-4） mRNA in peripheral blood mononuelear cells （PBMCs） were detected by real-time quantitative PCR. The serum IFN-γ and IL-4 concentrations were determined by enzyme-linked immunosorbent assays. Intrahepatic expressions of IFN-γ and IL4 were detected by imnmnohistochemical staining. Differences between groups were analyzed using non-parametric Kruskal-Walfis 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 （χ2 = 36. 259, 40. 822, 26. 321 and 31. 852, respectively, all P 〈 0. 05 ）. Serum and intrahepatie IFN-γ/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 imbalaneed Th1/Th2 ratio. With the degree of liver fibrosis exacerbations, Th1/Th2 eytokines drift into Th2 lymphocyte sub-cluster, which suggests that Th1/Th2 imbalance may be involved in the pathogenesis of CHB fibrosis.