通过调控滤泡性辅助性T细胞亚群研究蜈蚣败毒饮治疗银屑病模型鼠的机制

The Mechanism of Wugong Baidu Decoction in the Treatment of Psoriasis Mice Model Based on Regulating the Follicular Helper T Cell

目的观察蜈蚣败毒饮对银屑病鼠外周血滤泡辅助性T细胞(Tfh)表达量及对Tfh亚群Tfh1/Tfh2/Tfh17的影响,同时结合银屑病鼠皮损指数的变化量,分析Tfh及其亚群与皮损疗效之间的相关性。方法将银屑病鼠随机分为空白组、模型组、甲氨蝶呤对照组、蜈蚣败毒饮低、中、高剂量组,共6组。经咪喹莫特造模及小鼠灌胃后,观察银屑病模型鼠的皮损指数,而后进行含药外周血单个核细胞(PBMC)制备,对各组样本采用流式细胞法以CD4^(+)CXCR3^(+)CCR6^(-)作为细胞表面标记检测Tfh1细胞的表达量,以CD4^(+)CXCR3^(-)CCR6^(-)作为细胞表面标记检测Tfh2细胞的表达量,以CD4^(+)CXCR3^(-)CCR6^(+)作为细胞表面标记检测Tfh17细胞的表达量,得出了各组样本中Tfh细胞及各亚群Tfh1、Tfh2、Tfh17的数目和表达率,双变量回归分析法对Tfh及其亚群与皮损指数之间的相关性进行分析。结果各药物组均能降低银屑病模型鼠的皮损指数(P<0.05),以蜈蚣败毒饮中、高剂量组和甲氨蝶呤组的作用最为显著(P<0.01);蜈蚣败毒饮中、高剂量组和甲氨蝶呤组可下调Tfh细胞表达(P<0.05),下调Tfh17细胞表达(P<0.05);Tfh及Tfh17细胞表达量与皮损指数变化量(△MPASI)间呈负相关,Tfh1及Tfh2细胞表达量与皮损指数变化量(△MPASI)间无明显相关性。结论蜈蚣败毒饮可能通过下调咪喹莫特诱导银屑病鼠模型外周血中Tfh17细胞的表达,来改善银屑病模型鼠的皮损。

Objective To observe the effect of Wugong Baidu decoction on Tfh expression level and Tfh subgroup Tfh1/Tfh2/Tfh17 in psoriatic rats,and to analyze the correlation between Tfh and its subgroupbased on the skin lesion index of psoriasis model mice.Methods Psoriatic rats were randomly divided into blank group,model group,methotrexate group,low-dose group,medium-dose group,and high-dose group.After imiquimod and intragastric administration,the skin lesion index of psoriasis model mice was observed,Flow cytometry was used to detect the expression of Tfh1 cells with CD4^(+)CXCR3^(+)CCR6^(-)as a cell surface marker,the expression of Tfh2 cells with CD4^(+)CXCR3^(-)CCR6^(-)as a cell surface marker and the expression of Tfh17 cells with CD4^(+)CXCR3^(-)CCR6^(+)as a cell surface marker.The number and expression rate of Tfh cells and subsets of Tfh1,Tfh2 and Tfh17 in each group were obtained.Bivariate regression analysis was used to analyze the correlation between Tfh and its subsets and skin lesion index.Results Each drug group was able to reduce the skin lesion index of psoriasis model mice(P<0.05).The effects of high dose group and medium dose group of Wugong Baidu decoction and methotrexate group were the most significant(P<0.01);high dose group and medium dose group of Wugong Baidu decoction and methotrexate group could down-regulate the expression of Tfh cel land Tfh17 cell(P<0.05).The expression levels of Tfh and Tfh17 cells were negatively correlated with the change of skin lesion index(△MPASI),while there was no significant correlation between the expression levels of Tfh1 and Tfh2 cells and the change of skin lesion index(△MPASI).Conclusion Wugong Baidu decoction can effectively improve the skin lesions of psoriasis mice model by down-regulating the expression of Tfh cells in peripheral blood of the psoriatic mouse model induced by imiquimod.Among the three Tfh subgroups,Tfh17 cells are mainly involved in the occurrence and development of psoriasis.