慢性乙型肝炎不同脾虚兼证与外周血树突状细胞表型及功能的关系

目的： 研究慢性乙型肝炎（chronic hepatitis B, CHB）患者不同脾虚兼证与外周血树突状细胞（dendritic cell, DC）表型及功能的关系。方法：CHB患者60例，按中医证候诊断标准分为脾虚肝郁、脾虚湿热和脾肾两虚组，每组20例，10名健康人为正常对照。分离各组外周血单个核细胞，体外诱导培养DC，流式细胞仪测定DC表型，酶联免疫吸附检测法测定DC分泌白细胞介素10（interleukin-10, IL-10）浓度，同步检测患者生化指标及乙肝病毒的脱氧核糖核酸（hepatitis B virus DNA, HBV-DNA）水平。结果：CHB患者DC体外诱导增殖能力较正常人下降，DC成熟表面标记物显著低于正常人（P〈0.05）。不同证候CHB患者DC表面标记物CD80、CD86和CD1a表达阳性率均低于正常人（P〈0.05）。脾虚肝郁证患者CD80、CD1a和HLA-DR表达阳性率显著高于脾肾两虚证（P〈0.05），脾虚湿热证患者CD1a表达阳性率显著高于脾肾两虚证（P〈0.05）。CHB患者DC培养上清中IL-10浓度显著高于正常人（P〈0.05）。脾肾两虚证患者DC培养上清中IL-10含量高于脾虚肝郁证（P〈0.05）。结论：在CHB发病过程中，不同脾虚兼证CHB患者DC的表型及功能间存在差异，提示中医证候分类与机体免疫功能之间存在一定相关性。

Relationship between the phenotypes and functions of peripheral blood dendritic cells and the different spleen deficiency syndrome types in patients with chronic hepatitis B

Objective： To study the phenotypes and functions of dendritic cells （DCs） derived from peripheral blood monocytes of chronic hepatitis B （CHB） patients with different traditional Chinese medicine （TCM） syndrome types, and to explore the relationship between TCM syndrome type and DC functions.
Methods： Sixty CHB patients were included in this study. All the CHB patients were divided into spleen deficiency and liver stagnation, spleen deficiency and dampness-heat and deficiency of both spleen and kidney groups according to TCM syndrome diagnosis standard. There were 20 cases in each group, and ten healthy people were included as normal control. The volunteer’s peripheral blood was collected for monocyte separation, biochemical test and hepatitis B virus DNA loads detection. DCs were induced and isolated from peripheral blood monocytes, and then the expressions of surface markers CD80, CD86, CD1a and HLA-DR were detected by flow cytometric analysis method. Interleukin-10 （IL-10） production of the DCs was quantified by enzyme-linked immunosorbent assay.
Results： The proliferation of DCs in the CHB patients was slower than that in the healthy volunteers （P〈0.05）. The expressions of DC surface molecules such as CD80, CD86, and CD1a were obviously decreased in the CHB patients as compared with those in the healthy volunteers （P〈0.05）. More over, expressions of DC surface molecules were different among CHB patients with different TCM syndrome types. The positive expressions of CD80, CD1a, and HLA-DR in the CHB patients with spleen deficiency and liver stagnation were obviously higher than those in the CHB patients with deficiency of both spleen and kidney （P〈0.05）, and the CD1a expression in the CHB patients with spleen deficiency and dampness-heat was higher than that in the CHB patients with deficiency of both spleen and kidney （P〈0.05）. In DC culture supernatant, the IL-10 concentration of the CHB patients with deficiency of both spleen and kidney was higher than that of the CHB patients with spleen deficiency and liver stagnation （P〈0.05）, and the IL-10 concentrations of the CHB patients with different TCM syndrome types were higher than that of the healthy volunteers （P〈0.05）.
Conclusion： During the pathogenic course of CHB, the phenotypes and functions of DCs are different in CHB patients with different TCM syndrome types. It suggests that there is a correlation between TCM syndrome type and body immunity function.