同型半胱氨酸对C17.2小鼠神经干细胞的影响及作用机制  

Effects of homocysteine on neural stem cells and its underlying mechanism

作　　者：王丹[1] 阮妙华[1] 周爱华[1] 钱燕 陈亦明[2] 

机构地区：[1]温州医科大学附属第一医院新生儿科,325000 [2]温州医科大学附属第二医院肝胆外科

出　　处：《浙江医学》2017年第23期2071-2074,2078,共5页Zhejiang Medical Journal

基　　金：国家自然科学基金项目(81701485);浙江省医药卫生科技计划项目(2017KY452);温州市科技局项目(2017Y0578)

摘　　要：目的探讨同型半胱氨酸(Hcy)对C17.2小鼠神经干细胞(NSCs)的影响及可能的分子机制。方法培养C17.2小鼠NSCs,分为对照组、0.25m M Hcy组、0.25m M Hcy+5mM N-乙酰半胱氨酸(NAC)组;加入相应药物培养24h后,采用CCK-8法检测各组NSCs生长活力,流式细胞术和Caspase-3法检测Hcy对NSCs凋亡的影响,彗星实验检测Hcy对NSCs DNA的损伤程度,H2DCF-DA染色检测NSCs的氧自由基(ROS)水平。结果 Hcy能诱导细胞ROS产生和DNA损伤,从而导致NSCs凋亡增加。相反,NAC可降低Hcy诱发的ROS产生,明显改善DNA损伤,提高细胞存活率。结论 Hcy能诱导NSCs ROS产生和DNA损伤,并导致NSCs凋亡;而减少Hcy引发的ROS产生可以明显改善DNA损伤并提高细胞存活率。Objective To investigate the effects of homocysteine(Hcy) on C17.2 mouse neural stem cells(NSCs), and to explore its possible mechanism. Methods C17.2 mouse NSCs were cultured and divided into blank control group, Hcy group and Hcy+ N-acetylcysteine(NAC) group. The growth activity of NSCs was detected by CCK-8 method. The apoptosis of NSCs was assessed by flow cytometry analysis and caspase-3 method. Comet assay was used to evaluate the DNA damage of NSCs.The intracellular reactive oxidative species(ROS) level was detected by H_2DCF-DA staining method. Results Hcy evoked ROS production and induced cell DNA damage, leading to an increase in NSCs apoptosis. On contrary, NAC decreased Hcy-evoked ROS production and significantly ameliorated DNA damage and improved NSCs survival. Conclusion Hcy may play a negative role in NSCs survival via inducing DNA damage by oxidative stress.

关 键 词：同型半胱氨酸 DNA损伤 氧化应激 神经干细胞 

分 类 号：R363[医药卫生—病理学]