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目的:研究桑色素(morin)对小鼠T淋巴细胞活化、增殖和细胞周期的影响。方法:以刀豆蛋白A(ConA)刺激培养的淋巴结来源的小鼠淋巴细胞,再以不同终浓度的morin与T细胞共培养,利用流式细胞术(FCM),检测早期T细胞活化的标志CD69分子的表达,以羧基荧光素双醋酸盐琥珀酰脂(CFDA-SE)染色检测T细胞的增殖;以碘化丙锭(PI)染色分析T细胞的细胞周期。结果:小鼠T细胞培养6 h后,未经ConA刺激的对照组中CD69+T的细胞比率为(2.97±0.12)%,经ConA刺激的CD69+T细胞的比率明显增高,达到(72.52±0.66)%,与对照组相比差别明显(P<0.01)。终浓度为25、50、100μmol/L的morin均下调CD69+T细胞的比率,其中,100μmol/L的morin抑制作用最强,为(48.95±0.81)%,与对照组比较具有统计学意义(P<0.01)。CFDA-SE染色分析显示,ConA组培养48 h和72 h的T细胞的增殖指数(PI)分别为(1.58±0.04)和(1.95±0.02),各浓度的morin对ConA刺激的T细胞增殖,具有明显地抑制作用,以100μmol/L的morin抑制作用最明显。培养48 h的ConA组T细胞的PI为(1.02±0.02)、培养72 h的ConA组T细胞的PI为(1.03±0.01),与相应时间的对照组比较,均有统计学意义(P<0.01)。PI染色后流式细胞术分析的结果表明,ConA组处于S期的T细胞的比率为(27.05±0.39)%,显著高于对照组的比率(5.10±0.07)%。morin组中S期的细胞比率较高。结论:Morin可显著抑制ConA刺激的T细胞活化及增殖;其对增殖的抑制作用主要表现为S期的细胞的阻滞。
Objective: To study the effects of morin on the activation, proliferation and cell cycle of mouse T lymphocytes. Methods: The lymph node-derived mouse lymphocytes were stimulated with concanavalin A (ConA) and then co-cultured with different final concentrations of morin and T cells. Flow cytometry (FCM) was used to detect early T cell activation markers. The expression of CD69 molecule was detected by carboxyfluorescein diacetate succinyl ester (CFDA-SE) staining, and the cell cycle of T cells was analyzed by propidium iodide (PI) staining. RESULTS: After 6 h culture of mouse T cells, the ratio of CD69+ T cells in the control group without ConA stimulation was (2.97±0.12)%, and the ratio of CD69+ T cells stimulated by ConA was significantly higher, reaching (72.52±) 0.66)%, significantly different from the control group (P<0.01). The final concentrations of 25, 50, and 100 μmol/L of morin down-regulated the ratio of CD69+ T cells, of which 100 μmol/L of morin had the strongest inhibitory effect (48.95±0.81)%, which was statistically significant compared with the control group ( P<0.01). CFDA-SE staining analysis showed that the proliferation index (PI) of T cells in the ConA group at 48 h and 72 h was (1.58±0.04) and (1.95±0.02), respectively, and the concentration of morin stimulated proliferation of Con A-stimulated T cells. Obvious inhibitory effect, the inhibition of morin with 100μmol/L is the most obvious. The PI of the ConA group cultured for 48 h was (1.02±0.02), and the PI of the ConA group cultured 72 h (1.03±0.01) was statistically significant compared with the control group at the corresponding time (P< 0.01). The results of flow cytometry analysis after PI staining showed that the ratio of T cells in the S phase of the ConA group was (27.05±0.39)%, which was significantly higher than that of the control group (5.10±0.07)%. The proportion of cells in the S phase in the morin group was high. Conclusion: Morin can significantly inhibit the activation and proliferation of T cells stimulated by ConA. The inhibitory effect of Morin on the proliferation of T cells is mainly arrested by S phase cells.