基于树突状细胞表型及T淋巴细胞功能对HBV相关慢加急性肝衰竭患者中医虚实属性实质的分析

Analysis of TCM deficiency and excess attributes in patients with HBV- related acute- on- chronic liver failure based on phenotype of dendritic cells and function of T lymphocytes

目的归纳HBV相关慢加急性肝衰竭(HBV-ACLF)的中医虚实属性,分析其与树突状细胞(DC)表型及T淋巴细胞亚群的关系。方法收集2012年3月^(-1)1月于湖南中医药大学第一附属医院就诊的30例HBV-ACLF患者的基本资料、分期、中医证候等,归纳其中医虚实属性,分为实证组和虚证组;另选同期健康研究生志愿者10例作为对照组。应用流式细胞仪检测外周血CD3+、CD4+、CD8+T淋巴细胞百分比、CD4+CD25highCD127low表达率,以及外周血单个核细胞体外分离诱导培养的DC表型CD1α、HLA-DR、CD80、CD83、CD86表达率。计量资料符合正态分布的组间比较采用t检验,非正态分布的组间比较采用Wilcoxon秩和检验;计数资料组间比较用χ~2检验。结果与健康对照组相比,HBV-ACLF患者外周血CD3+、CD4+、CD8+T淋巴细胞百分比、CD4+与CD8+细胞比值及DC表型CD1α、HLA-DR、CD80、CD83、CD86的表达率均显著降低,CD4+CD25highCD127low表达率显著增高,差异均有统计学意义(P值均<0.01)。HBV-ACLF患者中,与实证组比较,虚证组外周血CD3+、CD4+T淋巴细胞百分比、CD4+与CD8+细胞比值及CD4+CD25highCD127low表达率均显著降低,差异均有统计学意义(P值均<0.05);与虚证组比较,实证组患者DC表型CD1α、CD83、CD86分子表达率均显著增高,差异均有统计学意义(P值均<0.05)。结论 HBV-ACLF虚证组、实证组患者均存在T淋巴细胞亚群功能下降及DC活化功能不足,其在虚证组更为显著;DC表型及T淋巴细胞亚群等免疫指标可以作为HBV-ACLF患者中医虚实属性的客观参考指标。

Objective To summarize the traditional Chinese medicine（ TCM） deficiency and excess attributes in patients with HBV- related acute- on- chronic liver failure（ HBV- ACLF）,and to analyze its association with the phenotype of dendritic cells（ DCs） and T lymphocyte subsets. Methods The basic information,stages,and TCM syndromes of 30 patients who were diagnosed with HBV- ACLF in The First Affiliated Hospital of Hunan University of Chinese Medicine from March to November,2012 were collected,and according to their deficiency and excess attributes,they were divided into excess group and deficiency group. Ten healthy volunteers were enrolled as the control group. Flow cytometry was used to measure the percentage of CD3＋,CD4＋,and CD8＋T lymphocytes in peripheral blood and the expression rate of CD4＋CD25highCD127lowcells,as well as the expression rates of DCs with phenotypes of CD1α,HLA- DR,CD80,CD83,and CD86 which were isolated from peripheral blood mononucleated cells,induced,and cultured in vitro,and its association with TCM deficiency and excess attributes. The t- test was used for comparison of normally distributed continuous data between groups,the Wilcoxon rank sum test was used for comparison of continuous data which were not normally distributed between groups,and the chi- square test was used for comparison of categorical data between groups. Results Compared with the healthy control group,the HBV- ACLF patients showed significant reductions in the percentages of CD3＋,CD4＋,and CD8＋T lymphocytes in peripheral blood and the expression rates of DCs with phenotypes of CD1α,HLA- DR,CD80,CD83,and CD86,as well as a significant increase in the expression rate of CD4＋CD25highCD127lowcells（ all P〈0. 01）. Compared with the excess group,the deficiency group showed significant reductions in the percentages of CD3＋and CD4＋T lymphocytes and the expression rate of CD4＋CD25highCD127lowcells（ all P〈0. 05）. Compared with the deficiency group,the excess group showed significant increases in the expression rates of DCs with phenotypes of CD1α,CD83,and CD86（ all P〈0. 05）. Conclusion The HBV- ACLF patients in deficiency and excess groups experience reduced functions of T lymphocyte subsets and insufficient activation function of DCs,which are more obvious in the deficiency group. The immune indices including phenotype of DCs and T lymphocyte subsets can be used as objective reference indices for determining TCM deficiency and excess attributes in HBV- ACLF patients.