尼古丁对脐带间充质干细胞微观形貌及增殖活性的影响  

The effects of nicotine on ultrastructure and proliferation of umbilical cord mesenchymal stem cells

作　　者：钟启[1] 覃永亮[2] 曾慧兰[3] 刘爽[1] 张青[1] 

机构地区：[1]广东省第二人民医院血液科,广东广州510317 [2]江门市人民医院血液科,广东江门529300 [3]暨南大学附属第一医院血液科,广东广州510630

出　　处：《中国热带医学》2015年第8期912-916,共5页China Tropical Medicine

基　　金：国家自然科学基金青年基金项目资助(No.81400168);广州市健康医疗协同创新重大专项基金项目资助(No.201400000003-1)

摘　　要：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 explore the effects of nicotine on morphology, cell cycle, proliferation and apoptosis of umbilical cord mesenehymal stem cells （MSCs）. Methods MSCs was treated with multiple concentration of nicotine, the variety of morphology was recorded by phase microscope and atomic force microscope. Cell growth was examined by MTT at 24h, 48h, 72h. Cell cycle and apoptosis were measured by flow cytometer at 24h. Results Upon nicotine treatment, the morphological changes of MSCs including cellular pyknosis, increased vacuolations, hollows and microvillis in the surface of cell membrane were noticed. The G0/G1 phase is dramatically increased（P〈0.05）, accompanied by gradually reduction of G2 and S phase by nicotine stimulation in a dose-dependent manner（P〈0.01）. Cell proliferation was inhibited by nicotine in a dose- and time-dependent manner. Compared with control group cell apoptosis rates in experimental groups were greatly increased by nicotine treatement for 24 hrs at 0.5, 1, 1.5 mg/mL. The cell apoptosis rate of cell treated with nicotine at 0.Stag/ mL was 4.867±0.404 % and （3.300±0.400）% in control group without significant difference （P〉0.05）.The cell poptosis rates of cells treated with nicotine at lmg/mL and 1.Smg/mL were （ 10.333±0.961 ） % and （44.367±3.612） %, respectively, all showed significant difference compared to that in control group （P〈0.01）. Conclusions Nicotine stimulation can alter MSCs morphology, ultrastrueture and induce early apoptosis, as well as inhibit cell cycle, proliferation and promote cell apoptosis.

关 键 词：尼古丁 间充质干细胞 增殖 凋亡 微观形貌 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]