健脾为主辨证治疗对慢性乙型肝炎患者外周血树突状细胞表型功能的影响

摘要

目的:研究健脾为主辨证治疗对脾虚为主不同中医证候类型慢性乙型肝炎(CHB)患者,外周血树突状细胞(DC)表型、功能的影响.方法:CHB患者60例,中医诊断符合脾虚肝郁、脾虚湿热、脾肾两虚等证候,每组20例,10名健康人为正常对照组.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；ELISA法测定DC分泌IL-10含量；同步检测患者生化指标及HBV DNA水平；各组患者分别予健脾法为主的中药辨证治疗,疗程为3个月,治疗结束后重复指标检测.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)；不同中医证候类型患者DC表面标记物表达水平存在差异；其中脾虚肝郁组CD8O、CD1a、HLA-DR表达阳性率显著高于脾肾两虚组(P＜0.05)；CHB患者DC培养上清液中IL-10含量显著高于正常人(P＜0.05),脾肾两虚组DC培养上清液中IL-10含量显著高于中脾虚肝郁组(P＜0.05).治疗后各组DC成熟度及分泌功能均有不同程度改善,其中脾虚肝郁组CD80、CD86、CD1a表达率较治疗前显著提高(P＜0.05),脾虚湿热组、脾肾两虚组CD80、CD86、CD1a、HLA-DB表达率较治疗前显著提高,而IL-10含量显著降低(P＜0.05)；治疗后除脾虚湿热组CD80表达高于脾肾两虚组外,各型DC表面标记物与IL-10比较,差异无显著性意义.结论:在CHB发病过程中,脾虚为主不同中医证候患者DC的表型及功能低下并存在差异,中药辨证治疗可改善DC表型及功能.%Objective: To investigate effect of syndrome differentiation based on invigorating spleen on phenotypes and functions of dendritic cells (DCs) in chronic hepatitis B patients (CHB) with different traditional Chinese medicine (TCM) syndrome classification. Methods; Sixty CHB patients were included this study, according to TCM syndrome diagnosis standard. All the patients were divided into spleen deficiency and liver stagnation group, spleen deficiency and dampness-heat group, and deficiency of spleen and kidney group, each group were twenty. Ten healthy people were included as normal control group. All the volunteers were collected perip heral blood for separating monocytes and biochemistry and HBV DNA loads detection. DCs were isolated and induced from peripheral blood monocytes, then the expression of surface markers, CD80, CD86, HLA-DR and CD1 a were detected by flow cytometric analysis. IL-10 production of the DCs quantified by ELJSA. Repeating all these items after three mouth treatment Results: DCs of CHB patients proliferated slower than those of the healthy controls. Compared with normal population the expression of DCs surface molecules obviously descend in CHB groups. More over expression of DC surface molecules were different among different TCM syndrome classification. Expressions of CD80, Cdla, HLA-DR in spleen deficien-cy and liver stagnation group were obviously higher than deficiency of spleen and kidney group ( P < 0.05) . In DCs culture supernatant, content of IL-10 was higher in deficiency of spleen and kidney group than spleen deficiency and liver stagnation group ( P < 0. 05), and all the CHB groups was higher than normal control group ( P < 0. 05 ) . But the maturity and function had been improved after three mouth treatment In spleen deficiency and liver stagnation group expressions of CD80, CD86 and Cdla were higher obviously than before (P < 0.05 ) , In spleen deficiency and dampness-heat and deficiency of spleen and kidney groups expressions of CD80, CD86, Cdla, HLA-DR were higher obviously than before (P<0.05) . While the level of IL-10 were lower than before in each CHB group (P <0.05) . Except for expressions of CD80 in spleen deficiency and dampness-heat was higher obviously than deficiency of spleen and kidney group, there were no obviously difference on the other items after three mouth treatment Conclusion; In CHB patients the phenotypes and functions of DCs are defected and different in different syndrome classification of TCM, and syndrome differentiation and treatment based on invigorating spleen can improve them.