目的:研究桑色素(morin)对小鼠T细胞增殖、细胞周期和腹腔巨噬细胞分泌一氧化氮(nitrogen mon-oxidum,NO)的影响。方法:以佛波醇酯(phorbol 12,13-dibutyrate,PDB)和离子霉素(ionomycin,Ion)联合刺激淋巴结来源的小鼠T淋巴细胞,再以不同终浓度的morin与上述细胞共培养,利用流式细胞术(FCM),以羧基荧光素双醋酸盐琥珀酰脂(carboxyfluorescein diacetate succinimide ester,CFDA-SE)染色检测T细胞的增殖;以碘化丙锭(pro-pidium iodide,PI)染色分析T细胞的细胞周期;脂多糖(Lipopolysaccharide,LPS)刺激小鼠腹腔巨噬细胞,与不同浓度morin共培养,以Griess反应检测上清液中的NO含量。结果:CFDA-SE染色分析显示,PDB+Ion组的增殖指数(proliferation index,PI)为1.70±0.05,各浓度的morin对PDB+Ion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,PI为1.03±0.01(P〈0.01)。细胞周期的分析结果表明,morin组中处于S期的细胞比率较高,与PDB+Ion组相比有明显差异。Griess反应检测NO含量的结果显示,各浓度组的morin均抑制LPS刺激巨噬细胞产生NO,100μmol/L的morin可将LPS刺激巨噬细胞产生NO为(12.89±0.11)μmol/L降低为(7.25±0.05)μmol/L,抑制作用最强。结论:Morin可显著抑制PDB+Ion刺激的T细胞增殖;其对增殖的抑制作用主要表现为S期的细胞的阻滞;对
Aim :To investigate the effects of morin on the proliferation and cell-cycle of mouse T lymphocytes and nitric oxide (NO) secretion by macrophages. Methods: Murine lymph node-derived T lymphocytes were separated and stimulated with phorbol 12,13-dibutyrate (PDB) plus ionomycin (Ion), followed by co-culture with different final concentrations of morin in different experimental groups. Flow cytometry (FCM) was employed to detect the proliferation [ carboxylfluorescein diacetate, succinimide ester (CFDA-SE) staining ] and cell-cycle [ propidium iodide (PI) staining ] of T cells. Macrophages isolated from irrigating solution were stimulated with Lipopolysaccharide (L/X3), the same experimental groups were set as described previously. Griess reaction was used to detect the production of NO in the cultivated supernate fluid. Results: CFDA-SE staining showed that PDB + Ion group proliferation index (PI) value was 1.70±0. 05,morin could decrease the PI value at final concentration being 25, 50 and 100 μmoL/L,with a peak at 100 μmoL/L morin ( 1.03±+0.01, P 〈0.01 ). FCM analysis of PI staining implied that the 25 and 50 μmol/L morin groups showed higher S phase cell rates than that in PDB + Ion group. Griess reaction analysis revealed that the morin could significantly suppressed the NO production of macrophages stimulated with LPS, 100 μmoL/L morin could drop off the NO production from 12.89 ± 0. 11 of LPS group to 7.25 ± 0. 05. Conclusion: Morin can significantly proliferation of murine T lymphocytes stimulated by PDB plus Ion, in which the S phase lagging may serve as one of the major mechanisms. Morin could decrease the NO output of peritoneal macrophages.