桑色素对小鼠T淋巴细胞体外活化、增殖和细胞周期的影响

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。

The effect of morin on activation, proliferation and cell-cycle of murine T lymphocytes in vitro

AIM: To discover the effects of morin on the activation, proliferation and cell-cycle of murine T lymphocytes in vitro. METHODS: Murine lymph node-derived T lymphocytes were separated and stimulated with concanavalin A (ConA) and different experimental groups were set by co-cultured with morin of different final concentration. Flow cytometry (FCM) was used to detect the activation, proliferation [carboxylfluorescein diacetate, succinimide ester (CFDA-SE) staining] and cell-cycle [propidium iodide(PI) staining] of T cells. RESULTS: After 6 h time of culture in vitro, the rate of CD69(+) T cells in control group was (2.97+/-0.12)%, while it was significant higher in ConA group [(72.52+/-0.66)% (P<0.01)]. Morin could down-regulate this rate at final concentration being 25, 50 and 100 micromol/L, with a peak at 100 micromol/L morin [(48.95+/-0.81)% (P<0.01)]. CFDA-SE staining showed that at 48 h and 72 h, the proliferation indexes (PI) of T cells in ConA group were (1.58+/-0.04) and (1.95+/-0.02), respectively. Morin could significantly decrease the PI value at all experimental concentration, with the peak effect at 100 micromol/L morin, which the PI for 48 h was (1.02+/-0.02) and (1.03+/-0.01) for 72 h (P<0.01). FCM analysis of PI staining implied that the percentage of S phase cells in ConA group was (27.05+/-0.39)%, significantly higher than that in control group (5.10+/-0.07)%; and the 25 and 50 micromol/L morin groups showed higher S phase cell rates. CONCLUSION: Morin can significantly inhibit ConA stimulated activation and proliferation of murine T lymphocytes, in which the S phase lagging may serve as one of the major mechanisms.