温法干预对HBV相关慢加急性肝衰竭阳黄证患者外周血树突细胞功能的影响  

Effects of Wenfa on peripheral blood dendritic cells in patients with HBV related acute-on-chronic liver failure of Yang Huang syndrome

作　　者：马远 唐丹 孙克伟[1] MA Yuan;TANG Dan;SUN Ke-wei(Hunan University of Chinese Medicine(Changsha,410208),China;不详)

机构地区：[1]湖南中医药大学第一附属医院肝病科,湖南长沙410208

出　　处：《中西医结合肝病杂志》2023年第5期430-435,共6页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

基　　金：国家自然科学基金面上项目(No.81973833);湖南省研究生创新型课题(No.2020CX19)。

摘　　要：目的:研究温法干预对HBV相关慢加急性肝衰竭(HBV-ACLF)阳黄证患者外周血树突状细胞(DC)功能的影响。方法:分别选取8例HBV-ACLF阳黄证患者作为阳黄组和8名健康对照者,收集两组人员外周抗凝血,体外诱导培养成成熟的DCs。将两组人员的DCs细胞分为5组:正常组、模型组、温法干预组、去温法干预组和空白组。并用大鼠温阳解毒化瘀方(温法)及温阳解毒化瘀方减附子、白术方(去温法)含药血清干预模型组(HBV-ACLF阳黄证患者)DCs。流式细胞仪分析DCs表面CD80、CD86、CD83、HLA-DR表达及平均荧光强度;ELISA法检测DCs分泌细胞因子IL-12、IL-6、IL-10水平;混合淋巴细胞反应(MLR)检测DCs对T淋巴细胞增殖的影响。结果:与正常组DCs比较,模型组DCs表面CD80、CD86、CD83、HLA-DR表达及平均荧光强度明显降低,DCs分泌细胞因子IL-12、IL-10显著减少,IL-6增多,促T淋巴细胞增殖功能下降(P<0.01);与模型组比较,温法干预组DCs表面CD80、CD86、CD83、HLA-DR表达及平均荧光强度显著升高,DCs分泌细胞因子IL-12、IL-10显著增多,IL-6减少,促T淋巴细胞增殖功能提高。结论:温法干预能明显提高HBV-ACLF阳黄证患者外周血DCs细胞功能,有利于清除HBV病毒、恢复患者紊乱的免疫功能。Objective:To explore the effects of Wenfa on the function of dendritic cells(DC)in patients with HBV related acute-on-chronic liver failure(HBV-ACLF)of Yanghuang syndrome.Methods:A total of eight patients with HBV-ACLF of Yanghuang syndrome and eight healthy people were selected.A total of 20 ml peripheral blood was collected by EDTA tube and peripheral blood mononuclear cells(PBMC)were isolated by the density gradient method.Then CD14+monocytes cells were isolated from PBMC by magnetic bead separation and were induced to mature DCs in vitro.Wenfa rat containing serum(Wenyang Jiedu Huayu decoction)and Qu Wenfa rat containing serum(Wenyang Jiedu Huayu decoction delete Fuzi,Baizhu)were used to intervene DCs.The expression and mean fluorescence intensity(MFI)of CD80,CD86,CD83,and HLA-DR on DCs were analyzed by flow cytometry.The IL-12,IL-6,and IL-10 secreted by DCs were detected by enzyme linked immunosorbent assay(ELISA).The effects of DCs on the proliferation of T lymphocytes were detected by mixed lymphocyte reaction(MLR).Results:The purity of DCs induced by CD14+monocytes cells in vitro was 96.4%.Compared with the normal group(healthy control),the expression and MFI of CD80,CD86,CD83 and HLA-DR on the surface of DCs in the model group was significantly decreased,the secretion of cytokines IL-12 and IL-10 in DCs were significantly decreased,IL-6 was increased,and the function of promoting T lymphocyte proliferation was decreased(P<0.01).Compared with the model group(HBV-ACLF Yanghuang syndrome group),the expression and MFI of CD80,CD86,CD83 and HLA-DR on the surface of DCs in Wenfa intervention group were significantly increased,IL-12 and IL-10 were increased,IL-6 was decreased,and the function of promoting T lymphocyte proliferation was improved(P<0.01);In Qu Wenfa intervention group,the expression and MFI of CD80,CD86 and CD83 on the surface of DCs were higher than model group,and the secretion of IL-12 and IL-10 were slightly increased,but there were no significant difference(P>0.05).Conclusion:Wenfa intervent

关 键 词：温法干预 乙型肝炎相关慢加急性肝衰竭 阳黄证 树突细胞 含药血清 

分 类 号：R575.3[医药卫生—消化系统]