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目的:分析连翘(FS)提取物对小鼠淋巴结T细胞的体外活化与增殖的影响,初步探讨其免疫抑制作用机制。方法:无菌分离小鼠淋巴结细胞,加入多克隆刺激剂刀豆蛋白A(ConA)进行刺激,利用荧光标记的单克隆抗体(mAb)染色结合流式细胞术(FCM),检测小鼠T淋巴细胞的表达的活化抗原CD69、CD25、CD71的表达情况;以羧基荧光素乙酰乙酸琥珀酰亚胺酯(CFDA-SE)染色,以FCM分析FS对淋巴细胞体外增殖的影响。结果:终浓度为40、80、160 mg/L的FS均对ConA刺激诱导的T细胞CD69、CD25和CD71的表达有降低作用(P<0.05)。CFDA-SE染色分析显示,上述浓度的FS对ConA诱导的小鼠T淋巴细胞增殖具有抑制作用(P<0.05)。结论:FS对ConA诱导的T细胞早、中、后期活化和体外增殖有抑制作用。
OBJECTIVE: To analyze the effect of extracts of Forsythia suspensa (FS) on the activation and proliferation of lymph node T cells in mice and to explore the mechanism of immunosuppression. METHODS: Mouse lymph node cells were aseptically isolated, stimulated with the polyclonal stimulator concanavalin A (ConA), and stained with a fluorescently labeled monoclonal antibody (mAb) combined with flow cytometry (FCM) to detect T lymphocytes in mice. The expression of activated antigens CD69, CD25, and CD71 expressed by cells was stained with carboxyfluorescein acetoacetate succinimide ester (CFDA-SE), and the effect of FS on lymphocyte proliferation in vitro was analyzed by FCM. RESULTS: FS with a final concentration of 40, 80, 160 mg/L had a decreased effect on ConA-induced expression of CD69, CD25, and CD71 in T cells (P<0.05). CFDA-SE staining analysis showed that the above concentration of FS inhibited Con A-induced proliferation of mouse T lymphocytes (P<0.05). Conclusion: FS can inhibit the activation of ConA-induced early, middle and late T cells and proliferation in vitro.