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目的:分析黄芩苷(BA)对小鼠T淋巴细胞体外增殖和细胞周期的影响,探讨其免疫抑制作用的机制。方法:无菌分离小鼠淋巴结,制备淋巴细胞悬液,以羧基荧光素双醋酸盐琥珀酰脂(CFDA-SE)染色,不同终浓度的BA预孵育4h,加入刀豆蛋白(ConA)或佛波醇酯类多克隆刺激剂PDB和离子霉素Ion进行刺激,以流式细胞术(FCM)结合荧光抗体染色技术检测黄芩苷对CD3+T细胞增殖率的影响;以碘化丙锭(PI)染色结合FCM分析其细胞周期分布。结果:BA(10、15、20、25mg/L)对ConA或PDB和Ion诱导的小鼠CD3+T细胞增殖均有明显的抑制作用(P<0.05),且呈浓度依赖性。对细胞周期分析表明,BA能使ConA或PDB和Ion刺激的小鼠淋巴细胞停滞于G0/G1期、S期,阻止其进入G2/M期,阻止作用随浓度的增加而增强。结论:BA对ConA或PDB和Ion诱导的小鼠CD3+T细胞增殖有明显抑制作用,能够将淋巴细胞阻滞在G0/G1期和S期,表现出明显的周期特异性。本研究提示黄芩苷有发展成免疫抑制剂的潜力。
Objective: To analyze the effect of baicalin (BA) on the proliferation and cell cycle of mouse T lymphocytes in vitro and to explore the mechanism of its immunosuppression. Methods: Lymph nodes were isolated aseptically and the lymphocyte suspension was prepared. The cells were stained with carboxyfluorescein diacetate succinate (CFDA-SE) and preincubated with BA at different final concentrations for 4 h. ConA or The effects of baicalin on the proliferation of CD3 + T cells were detected by flow cytometry (FCM) combined with fluorescent antibody staining. The effects of propidium iodide (PDI) and ionomycin Ion PI) staining and FCM analysis of cell cycle distribution. Results: BA (10,15,20,25 mg / L) significantly inhibited the proliferation of mouse CD3 + T cells induced by ConA or PDB and Ion (P <0.05) in a concentration-dependent manner. The cell cycle analysis showed that BA can make ConA or PDB and Ion-stimulated mouse lymphocytes arrest in G0 / G1 phase, S phase, prevent them from entering G2 / M phase, and the blocking effect increases with increasing concentration. CONCLUSION: BA can significantly inhibit the proliferation of mouse CD3 + T cells induced by ConA or PDB and Ion, and can arrest lymphocytes in G0 / G1 phase and S phase, showing obvious cycle specificity. This study suggests that baicalin has the potential to develop as an immunosuppressive agent.