慢性HBV感染不同免疫状态肝郁脾虚证与肝胆湿热证患者外周血树突状细胞功能研究

Study on the Functions of Peripheral Dendritic Cells in Chronic Hepatitis B Virus Infection Patients of Gan-depression Pi-deficiency Syndrome and Gan-Dan Damp-heat Syndrome under Different Immune States

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中文关键词: 慢性HBV感染肝郁脾虚证肝胆湿热证树突状细胞

英文关键词:chronic hepatitis B virus infection Gan-depression Pi-deficiency syndrome Gan-Dan damp-heat syndrome dendritic cell

基金项目:湖南省科技厅科技计划资助项目(No.2007FJ40);湖南省自然科学基金资助项目(No.09JJ6042);国家中医药管理局中医药科学技术研究专项资助项目(No.06-07JB17)

作者	单位
孙克伟	长沙市中心医院
彭建平	长沙市中心医院
何芳	湖南中医药大学第一附属医院传染科

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中文摘要:

目的观察慢性乙肝病毒(hepatitis B virus,HBV)感染不同免疫状态肝郁脾虚证和肝胆湿热证患者外周血树突状细胞(dendritic cells,DCs)的功能变化,了解慢性HBV感染两证型的免疫表达特点,为中医辨证提供客观化依据。方法选取40例慢性HBV感染患者,根据免疫状态分为两组,慢性乙型肝炎组(免疫清除状态)20例,其中肝郁脾虚证10例,肝胆湿热证10例;慢性HBV携带组(免疫耐受状态)20例,其中肝郁脾虚证10例,肝胆湿热证10例;另选取湖南中医药大学研究生志愿者体检健康者10名为健康对照组,分别对其外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)进行体外培养,以倒置显微镜和电镜观察DCs外部形态和超微结构,检测DCs的表面分子HLA-DR、CD80、CD86、CD1a的表达及白细胞介素-12(IL-12)水平,采用MTT法检测其同种异体混合淋巴细胞增殖能力以比较DCs在慢性HBV感染不同阶段两证型中的功能特点,分析两证型间DCs各指标的差异。结果与健康对照组比较,慢性HBV携带组两个证型患者CD86、CD80及HLA-DR表达率降低,慢性乙型肝炎组两个证型患者CD80表达率降低,差异均有统计学意义(P<0.05)。患者组肝郁脾虚证患者CD86、HLA-DR低于肝胆湿热证,慢性HBV携带组肝郁脾虚证CD80低于肝胆湿热证,差异均有统计学意义(P<0.05)。患者组IL-12及T淋巴细胞增殖能力低于健康对照组,差异有统计学意义(P<0.05)。慢性乙型肝炎组肝郁脾虚证患者IL-12及T淋巴细胞增殖能力低于肝胆湿热证,差异有统计学意义(P<0.05)。结论慢性HBV感染不同免疫状态肝郁脾虚证与肝胆湿热证患者外周血DCs功能存在差异。DCs表型及功能检测在疾病的同一阶段可为中医辨证分型提供客观化指标,反映两证型免疫表达的特点及免疫学内涵。

英文摘要:

Objective To observe the functions of peripheral dendritic cells(DCs) in chronic hepatitis B virus(HBV) infection patients of Gan-depression Pi-deficiency syndrome(GPS) and Gan-Dan damp-heat syndrome(GDS) under different immune states,thus to study the features of the immune expressions of the two syndromes in chronic HBV infection,providing objective evidence for Chinese medicine syndrome typing. Methods The 40 chronic HBV patients were randomly assigned to two groups according to the immune state. Of them,there were 20 chronic HBV patients(under the condition of immune clearance;consisting of 10 patients of GPS and 10 of GDS) and 20 chronic HBV carriers(under the condition of immune tolerance;consisting of 10 patients of GPS and 10 of GDS) . Besides,10 healthy graduate volunteers of Hunan University of Traditional Chinese Medicine were recruited as the healthy control group. Their peripheral blood mononuclear cells(PBMCs) were cultured in vitro. The exterior morphological features and ultrastructure were observed by inverted microscope and electron microscope. The expressions of HLA-DR,CD80,CD86,and CDla of the DCs surface were detected. The secretory levels of IL-12 in the supernate of DCs were detected by ELISA reagent kit. The proliferation capacities of allogeneic mixed lymphocyte were detected using MTT. The function features of DCs in the chronic HBV patients of two syndrome types under different immune states were compared,thus analyzing the difference of each index between the two syndrome types. Results Compared with the healthy control group,the expression rates of CD86,CD80,and HLA-DR decreased in the HBV carriers group(of the two syndrome types) ,showing statistical difference(P<0.05) . The expression rate of CD80 decreased in the HBV group(of the two syndrome types) ,showing statistical difference(P<0.05) . The expression rates of CD86 and HLA-DR were lower in the GPS group than in the GDS group. The expression rate of CD80 was lower in the GPS group than in the GDS group,showing statistical difference(P<0.05) . The proliferation capacities of IL-12 and T lymphocytes were lower in the HBV patients group than in the healthy control group(P<0.05) . The proliferation capacities of IL-12 and T lymphocytes were lower in the GPS group than in the GDS group,showing statistical difference(P<0.05) . Conclusions The functions of peripheral DCs in chronic HBV infection of patients of the GPS and the GDS under different immune states were different. The phenotype and function tests of DCs provided objective evidence for Chinese syndrome typing of chronic hepatitis B,thus reflecting the features of immune expressions of the two syndrome types and the immunology connotation.

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