目的 比较防己黄芪汤中活性成分在大鼠体内的药动学行为，并研究防己黄芪汤及其主要入血成分粉防己碱抑制刀豆球蛋白A（concanavalin A，ConA）诱导T淋巴细胞增殖的作用机制。方法 采用超高效液相色谱-串联质谱法（UPLC-MS/MS）检测大鼠ig不同剂量防己黄芪汤及粉防己碱后，入血成分在不同时间点的血药浓度，并分析其药动学行为。采用MTT法和³H-胸腺嘧啶核苷（³H-TdR）掺入法检测防己黄芪汤对T淋巴细胞毒性及增殖的抑制作用；采用ELISA法测定上清液中白细胞介素-17（interleukin-17，IL-17）、IL-2、γ-干扰素（γ-interferon，IFN-γ）水平；采用流式细胞仪检测调节性T细胞（regulatory T cell，Treg）数量。结果 防己黄芪汤在大鼠体内主要入血成分为防己诺林碱、木兰花碱、毛蕊异黄酮葡萄糖苷、甘草苷、黄芪甲苷、甘草次酸和粉防己碱。除防己诺林碱外，其他6个成分在体内的吸收具有剂量相关性。粉防己碱、防己诺林碱和甘草次酸在体内吸收速率慢，消除速率慢，驻留时间长[平均滞留时间（MRT）＞10 h、清除率（CL）＜8 L/(h∙kg)]；木兰花碱、毛蕊异黄酮葡萄糖苷和甘草苷在体内具有吸收和消除速度快、体内驻留时间短的特点[达峰时间（t_max）＜1 h、MRT＜5.2 h、CL＞50 L/(h∙kg)]。与单一成分给药相比，防己黄芪汤中的粉防己碱的吸收程度明显增加，而消除速率明显降低。防己黄芪汤可以抑制T淋巴细胞的增殖，还可降低细胞上清液中促炎因子IL-2、IL-17和IFN-γ水平（P＜0.05、0.01），增加Treg的分化数量（P＜0.05、0.01），防己黄芪汤主要入血成分之一粉防己碱可降低细胞上清液中IL-17水平并增加Treg分化数量（P＜0.05、0.01）。结论 君药防己和黄芪的特征成分是防己黄芪汤的主要入血成分；君药防己含有的粉防己碱与防己黄芪汤中其他成分存在协同作用。防己黄芪汤可抑制T淋巴细胞的增殖，其作用机制可能是通过调节Treg的增殖分化过程，使激活的免疫反应受到影响，而防己黄芪汤中的粉防己碱可能起到重要作用。

Objective To compare the pharmacokinetics of the active ingredients in Fangji Huangqi Decoction (防己黄芪汤, FHT), and study the mechanism of FHT and tetradrine (TET) on inhibiting concanavalin (ConA)-induced T lymphocytes proliferation. Methods An ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) technology was used to detect the plasma concentration of blood components at different time points after rats were ig different doses of FHT and TET, and the pharmacokinetic behavior was analyzed. The cytotoxicity and cell proliferation inhibitory activity of FHT on T cells was determined by MTT and ³H-thymidine (³H-TdR) incorporation method. The levels of interleukin-17 (IL-17), IL-2 and γ-interferon (IFN-γ) in the supernatant was determined by ELISA. The numbers of regulatory T cells (Treg) was detected by flow cytometry.Results Fangchinoline, magnoflorine, calycosin-7-glucoside, liquritin, astragaloside IV, glycyrrhetic acid and TET were the main components of FHT that entered the bloodstream in rats. Except for fangchinoline, the absorption of other six ingredients in the body were dose-dependent. TET, fangchinoline and glycyrrhetinic acid in the body had a slow absorption rate, a slow elimination rate and a long residence time [mean retention time (MRT) > 10 h, clearance rate (CL) < 8 L/(h∙kg)]. Magnoflorine, calycosin-7-glucoside and liquritin were rapidly absorbed, quickly eliminated, with a long residence time [T_max< 1 h, MRT < 5.2 h, CL > 50 L/(h∙kg)]. In comparison to single component administration, the absorption of TET in FHT was increased, while the elimination rate was decreased. FHT inhibited the proliferation of T lymphocytes, significantly reduced the levels of proinflammatory cytokines IL-2, IL-17 and IFN-γ in cell supernatant (P < 0.05, 0.01), increased the number of Treg differentiation (P < 0.05, 0.01). TET, one of the major compounds of FHT that entered the bloodstream, could decrease the content of IL-17 in cell supernatant and increase the number of Treg differentiation (P < 0.05, 0.01). Conclusion The characteristic components of the monarch drug Fangji (Stephaniae Tetrandrae Radix) and Huangqi (Astragali Radix) are the major blood entering components of FHT. Tetrandrine in monarch drug Stephaniae Tetrandrae Radix has a synergistic effect with other components in FHT. FHT can inhibit the proliferation of T lymphocytes, and its mechanism may be related to its actions in regulating the proliferation and differentiation process of Treg, affecting the activated immune response. Tetrandrine in FHT may play an important role.

上海市科委自然基金资助项目（21ZR1475200）