单核巨噬细胞极化在慢性活动性乙型肝炎中的表达及意义

摘要

目的：：研究慢性活动性乙型肝炎患者单核巨噬细胞的数量与功能变化。方法：随机选择51例慢性乙型肝炎患者(其中轻中度20例,重度31例)以及正常对照13例。采用Percoll分层液分离PBMCs,以CD14标记单核细胞,流式细胞仪检测PBMCs表面分子CD80、CD86、HLA-DR、CD163的表达；ELISA检测血清细胞因子IL-10、IL-12与IL-23的水平；免疫组织化学染色检测CD68在肝脏的分布。结果：轻中度慢性乙型肝炎组、重度慢性乙型肝炎组CD80表达水平低于对照组。慢性乙型肝炎患者外周血单核细胞CD86的表达低于对照组,其中重度组与对照组比较有统计学差异( P<0.01)。慢性乙型肝炎患者HLA-DR的表达低于对照组,其中轻中度组与重度慢性乙型肝炎组间HLA-DR的表达水平差异有统计学意义(P<0.01)。此外,慢性乙型肝炎患者外周血单核细胞CD163的表达明显高于对照组( P<0.01)。慢性乙型肝炎患者肝脏汇管区CD68阳性细胞浸润增加,汇管区增大(P<0.05)。轻中度慢性乙型肝炎组、重度慢性乙型肝炎组与对照组血清IL-10表达水平之间两两比较差异显著,均具有统计学意义(P<0.01)。结论：慢性乙型肝炎患者巨噬细胞参与肝脏病变的发生,外周血中单核细胞存在M1型/M2型平衡失调,向M2型极化的现象,可能参与其慢性化发展。%Objective:To research the number and function of monocyte-macrophages in patients with chronic active hepatitis B. Methods:The 51 chronic viral hepatitis B( CHB) patients were selected randomly,which consisted of 20 cases of mild-moderate,31 cases of severe group and 13 cases of healthy controls. PBMCs were separated by percoll. Monocytes were tagged by CD14,the molecules CD80,CD86,HLA-DR and CD163 were detected by flow cytometry which expressed on the surface of PBMCs. Serum cytokine were detected for IL-10, IL-12 and IL-23 by ELISA. The distribution of CD68 was detected in the liver by immunohistochemical staining. Results:The expressions of CD80 for all chronic hepatitis B patients were lower than the controls respectively,no matter mild-moderate or even severe group. Similarly,the HBV patients expressed lower level of CD86 in the peripheral blood mononuclear cells when compared with the control group. Furthermore, there was statistically difference between the levels of CD86 in severe group compared with control group (P<0. 01). As the expression of CD80 and CD86,the levels of HLA-DR in the patents had also declined when compared with controls. While the HLA-DR levels in both the mild-moderate HBV hepatitis groups were statistically significant higher than the severe group (P<0. 01). Different from the above all,the expression of CD163 in all chronic HBV hepatitis was higher than the control group. The CD68 positive cells in chronic HBV patients were observed and infiltrated increasingly in portal area and hepatic lobules (P<0. 05). There were statistically significant differences of IL-10 levels between the mild-moderate group,severe group and the control group,respectively (P<0. 01). Conclusion:Macrophages have participated in the pathological lesions of liver in CHB patients,among peripheral blood mononuclear cells,the phenomena of imbalance between type M1/M2 and polarization to type M2 have been observed,which participated in the development of the chronicity of CHB.