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目的探讨褪黑素(melatonin,MLT)对新生儿脐带血单个核细胞(cord bloodmononuclear cells,CBMC)活化增殖的影响。方法运用相差显微镜及~3 H-TdR 掺入法观察 MLT 对CBMC形态学的影响及增殖的调节,并与 MLT 对成人血单个核细胞(peripheral blood mononuclearcells,PBMC)的影响进行比较。结果 CBMC 的~3H-TdR 掺入率随培养基中 MLT 浓度(50 pg/ml～50 ng/ml)的增加而增加,在5 ng/ml 效果最佳。在加入 MLT(5 ng/ml)的培养基中,经72 h 培养后,高倍光镜观察下见细胞较培养前明显密集,为较多体积增大的单个核细胞。CBMC 培养基中分别加入不同刺激物 MLT(5 ng/ml)、Interleukin-2(IL-2,50 ng/ml)、MLT+phytohemagglutinin(PHA,5μg/ml)、MLT+IL-2,其~3 H-TdR 掺人率(cpm)分别为114 327±52 863、16 087±9006、118 360±59 207和17 682±7391。与细胞悬液组(14 133±8688)比较,MLT 组和 MLT+PHA 组 CBMC 的~3 H-TdR 掺入率显著增加(t=5.9143,P<0.001;t=5.5078,P<0.001),IL-2组、MLT+IL-2组 CBMC 的~3H-TdR 掺入率无明显变化(t=0.4983,P>0.05;t=0.9839,P>0.05);而 MLT 组或 MLT+PHA 组 CBMC 的~3H-TdR 掺入率与培养基中仅加入 PHA 组(110 397±48 663)比较,差异无统计学意义(t=0.1730,P>0.05;t=0.3286,P>0.05)。各种刺激物加入培养基后,CBMC 的~3H-TdR 掺入率均明显高于 PBMC。结论 MLT 不但可诱导 PBMC 活化增殖,也可诱导 CBMC 的活化增殖,且其对 CBMC 的作用强于PBMC。
Objective To investigate the effect of melatonin (MLT) on the activation and proliferation of neonatal cord blood mononuclear cells (CBMC). Methods The effects of MLT on morphology and proliferation of CBMC were observed by phase contrast microscopy and ~ 3 H-TdR incorporation. The effects of MLT on the expression of peripheral blood mononuclear cells (PBMCs) were compared. Results The ~ 3H-TdR incorporation of CBMC increased with the increase of MLT concentration (50 pg / ml ~ 50 ng / ml) in culture medium, and the best effect was at 5 ng / ml. In the culture medium with MLT (5 ng / ml), after 72 h of culture, the cells were observed to be more dense than those before culture under the high light microscope, and the cells were more bulky. CBMC medium was incubated with different concentrations of MLT (5 ng / ml), Interleukin-2 (50 ng / ml), MLT + phytohemagglutinin (PHA, The 3 H-TdR incorporation rates (cpm) were 114 327 ± 52 863,16 087 ± 9006,118 360 ± 59 207 and 17 682 ± 7391, respectively. Compared with the cell suspension group (14 133 ± 8688), the ~ 3 H-TdR incorporation rate in CBMCs in MLT group and MLT + PHA group increased significantly (t = 5.9143, P <0.001; t = 5.5078, P <0.001) The incorporation rate of ~ 3H-TdR in CBMC in IL-2 group and MLT + IL-2 group had no significant difference (t = 0.4983, P> 0.05; t = 0.9839, P> 0.05) The incorporation rate of ~ 3H-TdR was no significant difference (P> 0.05); t = 0.3286 (P> 0.05). After adding various stimulants into the culture medium, the ~ 3H-TdR incorporation rate of CBMC was significantly higher than that of PBMC. Conclusion MLT can not only induce the activation and proliferation of PBMC, but also induce the activation and proliferation of CBMC, and its effect on CBMC is stronger than that of PBMC.