【摘要】 通过分离制备小鼠淋巴细胞;CCK-8法检测脱水淫羊藿素(AHI)、淫羊藿素(ICA)对淋巴细胞的毒性;流式细胞术(FCM)结合双色免疫荧光染色技术检测AHI、ICA对con A刺激T淋巴细胞早期活化标志CD69,中期活化标志CD25的表达情况;利用活性染料CFSE,结合流式细胞术分析AHI、ICA对T淋巴细胞增殖的影响;流式细胞术分析AHI、ICA对T淋巴细胞周期的影响;流式细胞术实时分析AHI、ICA对T淋巴细胞钙离子内流的影响;使用流式蛋白定量检测技术(CBA)检测AHI、ICA对T淋巴细胞分泌细胞因子的影响.结果显示AHI与ICA对活化的小鼠T淋巴细胞具有明显的免疫抑制作用,特别是ICA,能明显抑制经con A刺激的T淋巴细胞早中期活化、增殖、钙离子内流,并阻滞细胞周期在G0/G1期,以及分泌IL-6、IL-10、IFN-γ、TNF和IL-12p70细胞因子;ICA比AHI对活化的小鼠T淋巴细胞具有更明显的免疫抑制作用.更多还原

【Abstract】 Mouse lymphocytes were isolated.The drug toxicology of different concentrations of AHI/ ICA on lymphocytes was measured by CCK-8 assay.The expression of CD69 and CD25,which were the markers of activation of T lymphocytes,was measured by FCM combined with two-color immunofluorescent staining of cell surface antigen.The proliferation of T lymphocytes was measured by FCM combined with live dye CFSE.The cell cycle of T lymphocytes was measured by FCM.The Calcium influx of T lymphocytes was measured by FCM’s time function.CBA kit was used to detect the production of cytokines of lymphocytes stimulated by con A.AHI and ICA decreased proportions of CD69 and CD25 on con A-stimulated lymphocytes in a dose-dependent manner.Both reduced the proliferation of T lymphocytes,and the influx of Calcium.AHI and ICA caused a G0 / G1 cell cycle arrest on con A-activated lymphocytes.Furthermore,both downregulated the secretion of cytokines(IL-6,IL-10,IFN-γ,TNF,IL12p70) of con A-activated lymphocytes.Furthermore,ICA exhibited more immunosuppressive effects than AHI on mouse T lymphocytes.更多还原