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目的探讨尼古丁对脐带间充质干细胞(MSCs)形态学、细胞周期、细胞增殖及凋亡的影响。方法不同浓度尼古丁作用于MSCs,以相差显微镜及原子力显微镜(Atomic Force Microscope,AFM)观察形态学变化;MTT法分别于24h、48h、72h检测细胞增殖;流式细胞仪法于24 h检测细胞周期与细胞凋亡。结果尼古丁作用于MSCs后,细胞固缩,细胞膜表面形成空洞及凹陷,微绒毛样突起多见;细胞周期改变,G0/G1期的细胞比例明显增加(P<0.05),G2期及S期细胞比例则逐渐减少(P<0.01),周期阻滞随着尼古丁浓度递增而增加;细胞增殖受抑,呈时间剂量依赖性,凋亡率增高,0.5、1、1.5mg/m L的尼古丁作用MSCs 24h后,细胞凋亡率均高于对照组。0.5mg/m L组凋亡率(4.867±0.404)%与对照组(3.300±0.400)%比较,差异无统计学意义(P>0.05);1mg/m L组凋亡率(10.333±0.961)%、1.5mg/m L组凋亡率(44.367±3.612)%与对照组比较,差异均有统计学意义(P<0.01)。结论尼古丁使MSCs形态及超微结构发生早期凋亡改变,阻滞细胞周期,抑制细胞增殖且促进其凋亡。
Objective To investigate the effects of nicotine on the morphology, cell cycle, cell proliferation and apoptosis of umbilical cord mesenchymal stem cells (MSCs). Methods Nicotine was administered to MSCs at different concentrations. Morphological changes were observed by phase-contrast microscopy and atomic force microscopy (AFM). Cell proliferation was detected by MTT assay at 24, 48 and 72 h, respectively. Cell cycle was detected by flow cytometry at 24 h With apoptosis. Results After nicotine treatment on MSCs, cell shrinkage was observed and voids and invagination were observed on the surface of the cell membrane. Microvilli-like processes were more common. The cell cycle was significantly increased in the G0 / G1 phase (P <0.05) (P <0.01). The cycle arrest increased with the increase of nicotine concentration. The cell proliferation was inhibited in a dose-and time-dependent manner, and the apoptosis rate was increased. The effect of nicotine at 0.5,1,1.5mg / m L on MSCs After 24 h, the apoptosis rate was higher than that of the control group. The apoptosis rate in the group of 0.5mg / m L (4.867 ± 0.404)% was not significantly different from that in the control group (3.300 ± 0.400)% (P> 0.05). The apoptosis rate in the group of 1mg / m L was (10.333 ± 0.961) %, And the apoptosis rate in the group of 1.5mg / m L (44.367 ± 3.612)% compared with the control group, the difference was statistically significant (P <0.01). Conclusion Nicotine can change the morphological and ultrastructural changes of MSCs early, block cell cycle, inhibit cell proliferation and promote apoptosis.