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目的探讨中药注射剂(TCMI)对免疫细胞致敏相关表面分子表达的影响,为TCMI致敏性评价提供参考。方法将雌性BALB/c小鼠用随机区组法分为对照组和12个TCMI组(分别为鸦胆子组、艾迪组、香丹组、葛根素组、红花组、醒脑静组、喜炎平组、舒血宁组、鱼腥草组、生脉组、黄芪组、灯盏细辛组),每组7只小鼠。按照临床等效剂量于12个TCMI组小鼠左后肢足趾部各注射相应TCMI 50μL,对照组注射等体积磷酸盐缓冲液。5 d后处死动物,取左侧腘窝淋巴结制备淋巴细胞悬液,应用流式细胞术检测CD4+T细胞、CD8+T细胞、B细胞、CD11c+细胞及F4/80细胞比例,早期活化分子CD69、MHCⅡ和协同刺激因子CD86、CD80、CD40及CD44分子的表达。结果与对照组相比,鸦胆子组CD4+T细胞比例及CD69表达均减低,艾迪组CD8+T细胞比例及CD69表达均减少,但2组B细胞比例均明显增高而其CD69表达均减低;葛根素组、香丹组和喜炎平组B细胞比例均增高而其CD69表达均减低,葛根素组T细胞CD69表达也减低,香丹组和喜炎平组CD8+T细胞比例降低;醒脑静组CD8+T细胞比例及T、B细胞CD69表达均减少;鱼腥草组、灯盏细辛组CD8+T细胞与B细胞CD69表达减低;红花组仅CD8+T细胞比例降低。与对照组比较,鸦胆子组、艾迪组、香丹组CD11c+细胞和F4/80细胞比例均增高,红花组和葛根素组仅F4/80细胞比例升高;除舒血宁组和鱼腥草组外,其他各组CD4+T细胞表面MHCⅡ分子表达均明显增高;鸦胆子组、艾迪组和葛根素组CD11c+细胞CD40表达增高,醒脑静组CD11c+细胞CD80表达升高,香丹组和红花组CD11c+细胞CD86表达增高,F4/80细胞CD40表达减低。与对照组比较,香丹组和喜炎平组CD4+T细胞表面CD44分子表达增高而黄芪组表达减低;鸦胆子组、艾迪组、香丹组、红花组、喜炎平组和醒脑静组CD8+T细胞表面CD44分子表达均增高。结论不同种类TCMI引起的腘窝淋巴结免疫细胞表面分子变化不同,对其综合考察可以为TCMI致敏性评价提供参考和依据。
Objective To investigate the effect of TCMI on sensitization of immune cells to surface molecules and to provide a reference for the evaluation of TCMI sensitization. Methods Female BALB / c mice were randomly divided into control group and 12 TCMI groups (Brucea javanicae, Eddie group, Xiangdan group, Puerarin group, Honghua group, Xingnaojing group, Xiyanping group, Shuxuening group, Houttuynia group, Shengmai group, Astragalus group, Erigeron group), each group of 7 mice. In accordance with the clinical equivalent dose of 12 TCMI group mice left hind foot toe each injection corresponding TCMI 50μL, the control group injected with an equal volume of phosphate buffer. After 5 days, the animals were sacrificed and the left popliteal lymph nodes were prepared for lymphocyte suspension. The proportion of CD4 + T cells, CD8 + T cells, B cells, CD11c + cells and F4 / 80 cells were measured by flow cytometry. , MHC Ⅱ and costimulatory molecules CD86, CD80, CD40 and CD44. Results Compared with the control group, the ratio of CD4 + T cells and CD69 in Brucea javanicae group were decreased. The percentage of CD8 + T cells and CD69 expression in Edwards group were decreased. However, the proportion of B cells in both groups was significantly increased and the expression of CD69 was decreased ; The percentage of B cells in puerarin group, Xiangdan group and Xiyanping group were all increased while the expression of CD69 was decreased; the expression of CD69 in puerarin group was also decreased; the proportion of CD8 + T cells in Xiangdan group and Xiyanping group was decreased; The proportion of CD8 + T cells in Xingjingjing group and the expression of CD69 in T and B cells were decreased. The expression of CD69 in CD8 + T cells and B cells of Houttuynia cordata Thunb. And Erigeron breviscapus groups was decreased. The proportion of CD8 + T cells in Safflower group was decreased. Compared with the control group, the proportion of CD11c + cells and F4 / 80 cells in Brucea javanicae group, Aidi group and Xiangdan group were increased, while the proportion of F4 / 80 cells was only increased in safflower group and puerarin group; Houttuynia group, other groups of CD4 + T cell surface expression of MHC Ⅱ molecules were significantly increased; Brucea group, Aidi group and puerarin CD11c + cells CD40 expression increased, Xingnao Jing CD11c + CD80 expression increased, Xiangdan The expression of CD86 in CD11c + cells increased and the expression of CD40 decreased in F4 / 80 cells. Compared with the control group, the expression of CD44 on the surface of CD4 + T cells in Xiangdan group and Xiyanping group was increased, while the expression of Astragalus group was lower in Brucea group, Aidi group, Xiangdan group, Honghua group, Xiyanping group, Brain static group CD8 T cells surface expression of CD44 molecules were increased. CONCLUSION: The molecular changes of immune cell surface in popliteal lymph nodes caused by different kinds of TCMI are different. The comprehensive investigation can provide a reference and basis for the sensitization evaluation of TCMI.