中药注射剂对免疫细胞表面分子的影响

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TCMI50μL，对照组注射等体积磷酸盐缓冲液。5d后处死动物，取左侧胴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、MHC11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MHC11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引起的胴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。

Effect of traditional Chinese medicine injections on immune cell surface molecules

Objective To explore the effect of traditional Chinese medicine injections （TCMI） on expression of immune cell surface molecules and provide reference for evaluating the allergenicity of TCMI. Methods Using a random block design, the female BALB/c mice were divided into the control group and 12 kinds of TCMI groups[the Yadanzi （鸦胆子）, the Aidi （艾迪）, the Xiangdan（香丹）, the Gegensu（葛根素）, the Honghua （红花）, the Xingnaojing （醒脑静）, the Xiyanping （喜炎平）, the Shuxuening （舒血宁）, the Yuxingeao （鱼腥草）, the Shengmai （生脉）, the Huangqi （黄芪）, and the Dengzhanxixin （灯盏细辛） groups 1. Each group comprised 7 mice. According to the equivalent dose used in the clinical practice, TCMI 50 μl was subcutaneouslyinjected into the toe of left hind limb of each mouse in the TCMI groups, respectively. Equal volume of phosphate buffer solution was injected into the same site of each mouse in the control group. The mice were sacrificed on day 5 after injections, and the left popliteal fossa lymph node （PLN） was removed and the cell suspension were prepared. The ratio of CI4^＋ and CD8^＋T ceils, B cells, CD11 c ^＋cells, F4/80 cells and the expression of early activation molecules CD69, MHC 11 , co-stimulatory molecules CD86, CD80 and CIM0 were detected using flow cytometry. Results Compared with the control group, the ratio of CI4^＋ T cells and the expression of CD69 molecule decreased in the Yadanzi group ; the ratio of CD8^＋ T cells and the expression of CD69 molecule decreased in the Aidi group; however, the ratio of B cells increased and its expression of CD69 molecule decreased in the above-mentioned 2 groups ; the ratio of the B cells increased and expression of CD69 molecule decreased in the Gegensu, Xiangdan and Xiyanping groups, the expression of CD69 molecule decreased in the T cells in the Gegensu group; the ratio of CD8 ~ T cells decreased in the Xiangdan and Xiyanping groups, the ratio of CD8^＋ T ceils and the expression of CD69 molecule decreased in the Xingnaojing group ; the expression of CD69 in the CD8^＋ T and B cells in the Yuxingcao and Dengzhanxixin groups, the ratio of CD8^＋ T cells decreased in the Honghua group. Compared with the control group, the ratio of CD11 c^＋ and F4/80 cells increased in the Yadanzi, Aidi and Xiangdan groups; the ratio of F4/80 cells increased in the Honghua and Gegensu groups; the expression of MHC I1 molecule on the CD4^＋ T cell surface increased in 10 kinds of TCMI groups except the Shuxuening and Yuxingcao groups; the expression of CD40 molecule on the CDllc^＋ cell surface increased in the Yadanzi, Aidi and Gegensu groups; the expression of CD80 molecule on the CDllc^＋cell surface increased in the Xingnaojing group; the expression of CD86 molecule on the CD11 c ~ cell surface increased and the expression of CIMO molecule on the F4/80 cell surface decreased in the Xiangdan and Honghua groups. Compared with the control group, the expression of CD44 molecule on the CI4^＋T cell surface increased in the Xhmgdan and Xiyanping groups and decreased in the Huangqi group; the expression of CD44 molecule on the CD8^＋ T cell surface increased in the Yadanzi, Aidi, Xiangdan, Honghua, Xiyanping and Xingnaojing groups. Conclusion Different kinds of TCMI can cause various changes of immune cell surface molecules ; a comprehensive investigation of these molecules can provide scientific basis and reference for allergy evaluation of TCMI.