目的 考察复方扶芳藤合剂（CFM）对免疫抑制小鼠Th1/Th2细胞水平的调节作用及机制。方法 将SPF级Balb/c小鼠随机分为对照组，单给CFM低、中、高剂量（3.75、7.50、15.00 g·kg^-1）组，模型组，CFM低、中、高剂量（3.75、7.50、15.00 g·kg^-1）组，模型组和CFM低、中、高剂量组连续3 d ip 80 mg·kg^-1的环磷酰胺（CTX）建立免疫抑制模型，对照组和CFM单给药组不造模，仅ip等体积0.9%氯化钠溶液。造模结束后，每天ig给药1次，连续给药10 d，对照组和模型组ig等体积0.9%氯化钠溶液。末次给药24 h后取材，流式细胞术检测小鼠外周血Th1、Th2细胞水平及其表面活化（CD38、CD69）和耗竭［CD95、程序性死亡受体1（PD-1）］分子的表达水平，脾脏CD3+CD4⁺T细胞中γ干扰素（IFN-γ）、白细胞介素（IL）-2、IL-10细胞因子水平。结果 与对照组比较，模型组外周血Th1/Th2细胞向Th1型偏移，Th1细胞上CD38、CD95、PD-1的表达显著升高（P＜0.001），Th2细胞上CD38、PD-1的表达显著升高（P＜0.05、0.001），脾脏CD3+CD4⁺T细胞中IFN-γ的表达显著升高（P＜0.001）；与模型组比较，CFM 3.75 g·kg^-1组外周血Th1/Th2趋于平衡稳态，Th1细胞中CD38、PD-1的表达、Th2细胞CD38、PD-1的表达均显著下降（P＜0.05、0.01、0.001）；小鼠脾脏CD3+CD4⁺T细胞中IFN-γ和IL-2表达均显著下降（P＜0.05），IL-10的表达显著上升（P＜0.05、0.01、0.001）。单给CFM并不改变正常小鼠的Th1/Th2平衡及功能。结论 CFM改善免疫抑制小鼠外周血Th1/Th2细胞向Th1型偏移，对免疫抑制小鼠外周血Th1、Th2细胞的活化、耗竭具有正向调节作用，促进Th2细胞分泌IL-10，抑制Th1细胞分泌IFN-γ、IL-2。

Objective To investigate the regulatory effect of Compound Fufangteng Mixture (CMF) on Th1/Th2 cell level in immunosuppressed mice. Methods Balb/c mice were randomly divided into control group, single CFM low, medium, and high dose (3.75, 7.50, 15.00 g·kg^-1) groups, model group, CFM low, medium, and high dose groups, model group, and CFM low, medium, and high dose groups that were ip with 80 mg·kg^-1 of CTX for three consecutive days to establish an immunosuppressive model. The control group and CFM single-dose group were not subjected to modeling and were ip with an equal volume of 0.9% sodium chloride solution. After modeling was completed, the mice were ig given the drug once daily for 10 consecutive days. The control group and model group were ig given an equal volume of 0.9% sodium chloride solution. After the last drug administration, the mice were sacrificed and the peripheral blood Th1/Th2 cell levels, the surface activation (CD38, CD69) and exhaustion [CD95, programmed death receptor 1 (PD-1)] molecule expression levels, and the levels of cytokines interferon gamma (IFN-γ), interleukin (IL)-2, and IL-10 in the spleen CD3+CD4⁺T cells were detected by flow cytometry. Results Compared with the control group, the Th1/Th2 cells in the peripheral blood of the model group shifted towards the Th1 type. The expressions of CD38, CD95, and PD-1 on Th1 cells were significantly increased (P < 0.001), and the expressions of CD38 and PD-1 on Th2 cells were significantly increased (P < 0.05, 0.001). The expression of IFN-γ in the spleen CD3+CD4⁺T cells was significantly increased (P < 0.001). Compared with the model group, the Th1/Th2 in the peripheral blood of the CFM 3.75 g·kg^-1 group tended to be in a balanced and stable state. The expressions of CD38 and PD-1 on Th1 cells and the expressions of CD38 and PD-1 on Th2 cells were significantly decreased (P < 0.05, 0.01, 0.001). The expressions of IFN-γ and IL-2 in the spleen CD3+CD4⁺T cells of mice were significantly decreased (P < 0.05), and the expression of IL-10 was significantly increased (P < 0.05, 0.01, 0.001). The single administration of CFM did not change the Th1/Th2 balance and function of normal mice. Conclusion CFM improves the shift of Th1/Th2 cells in the peripheral blood of immunosuppressed mice towards the Th1 type. It has a positive regulatory effect on the activation and exhaustion of Th1 and Th2 cells in the peripheral blood of immunosuppressed mice, promotes the secretion of IL-10 by Th2 cells, and inhibits the secretion of IFN-γ and IL-2 by Th1 cells.

R285.5

国家自然科学基金资助项目(81860780);广西自然科学基金资助项目(2020GXNSFAA297162)