【摘要】 目的研究乙醇和乙酸铅联合染毒对胎鼠脑新皮质神经干细胞自我更新的影响及其机制。方法取SpragueDawley胎鼠脑新皮质的神经干细胞悬液,分设对照组、乙醇(25、50 mmol/L)单独染毒组、乙酸铅(0.01、0.1、1μmol/L)单独染毒组和乙醇+乙酸铅联合染毒组。培养48 h后,采用台盼蓝试验检测细胞存活率,采用流式细胞术检测细胞周期,采用qRT-PCR法检测神经干细胞标志物nestin和细胞周期相关基因p16和p21 mRNA表达。结果乙醇与乙酸铅联合染毒组胎鼠脑新皮质神经干细胞存活率下降,细胞周期延缓、G0/G1期阻滞和S期DNA合成抑制,nestin mRNA表达下降均存在协同效应(P<0.05);对p16和p21 mRNA表达水平增高亦呈一定协同作用(P<0.05)。结论乙醇和乙酸铅联合染毒能抑制胎鼠脑新皮质神经干细胞的自我更新,并且存在一定协同作用。更多还原

【Abstract】 Objective To investigate the combined toxic effects of ethanol and lead on the self-renewal of neural stem cells(NSCs) isolated from fetal rat cerebral neocortex and explore the action mechanisms. Methods Neural stem cell suspensions of Sprague-Dawley fetal rat cerebral neocortex were divided into control group,ethanol groups(25,50 mmol/L),lead groups(0.01,0.1,1 μmol/L) and combined groups. After 48 hours of culture,trypan blue exclusion test was used to examine cell viability,flow cytometry was used to examine cell cycle,and qRT-PCR was used to study the effects on mRNA expression of NSC marker nestin and cell cycle related genes p16 and p21. Results Ethanol and lead had synergistic effects in reducing cell viability, delaying cell cycle progression, arresting G0/G1-phase, inhibiting DNA synthesis of S-phase and,reducing nestin mRNA expression(P <0.05). Ethanol and lead also had a certain synergistic effect on increasing p16 and p21 mRNA expression(P <0.05). Conclusion Ethanol and lead have a certain synergistic effect on inhibiting self-renewal of NSCs isolated from fetal rat cerebral neocortex.更多还原