叶酸对缺氧大鼠神经干细胞Notch信号通路相关基因mRNA表达的影响

营养学报2011年第33卷第6期 567

叶酸对缺氧大鼠神经干细胞Notch信号通路

相关基因mRNA表达的影响

刘　欢，黄国伟，赵　琳，尤清菊　

（天津医科大学公共卫生学院营养与食品卫生学教研室，天津３０００７０）　

【摘　要】目的　研究叶酸对缺氧大鼠神经干细胞（ＮＳＣｓ）增殖及Ｎｏｔｃｈ信号通路相关基因ｍＲＮＡ表达的影响。方法　无血清培养基原代培养新生大鼠ＮＳＣｓ，细胞分为对照组（Ｃｏｎｔｒｏｌ），缺氧组（Ｈｙｐｏｘｉａ），叶酸组（Ｆｏｌａｔｅ）和叶酸缺乏组（Ｆｏｌａｔｅ－Ｄ）。除Ｃｏｎｔｒｏｌ组外，其余３组缺氧６ｈ。增殖第７日收集细胞，ＭＴＴ法检测细胞活力，实时荧光定量ＰＣＲ检测Ｎｏｔｃｈ１、Ｈｅｓ１／５、Ｍａｓｈ１和Ｎｇｎ１／２基因ｍＲＮＡ表达。结果　　Ｆｏｌａｔｅ组细胞活力高于Ｈｙｐｏｘｉａ组，Ｆｏｌａｔｅ－Ｄ组细胞活力低于其他３组（Ｐ＜０．０５）；Ｆｏｌａｔｅ组Ｎｏｔｃｈ１、Ｈｅｓ１／５ｍＲＮＡ表达高于Ｈｙｐｏｘｉａ组，Ｆｏｌａｔｅ－Ｄ组该３个基因ｍＲＮＡ表达低于其他３组（Ｐ＜０．０５）；Ｆｏｌａｔｅ组Ｎｇｎ１／２ｍＲＮＡ表达低于Ｈｙｐｏｘｉａ组（Ｐ＜０．０５）；Ｍａｓｈ１ｍＲＮＡ表达在各组间差别无统计学意义（Ｐ＞０．０５）。结论　叶酸能促进缺氧损伤ＮＳＣｓ增殖，上调Ｎｏｔｃｈ信号通路中Ｎｏｔｃｈ１和Ｈｅｓ１／５基因表达。　

［营养学报，２０１１，３３（６）：５６７－５７０］

关键词： 神经干细胞；叶酸；缺氧；Notch信号通路

中图分类号： R151.1 文献标识码：A 文章编号：0512-7955（2011）06-0567-04

EFFECTS OF FOLATE ON mRNA EXPRESSION OF GENES INVOLVED IN NOTCH SIGNAL

PATHWAY OF RAT NEURAL STEM CELLS AFTER HYPOXIA

LIU Huan，HUANG Guo-wei，ZHAO Lin，YOU Qing-ju

(Department of Nutrition and Food Hygiene，School of Public Health，Tianjin Medical University，Tianjin 300070，China)

【Abstract】 Objective To investigate the effects of folate on mRNA expression of genes involved in Notch signal pathway of rat neural stem cells（NSCs）after hypoxia. Method Neonatal rat NSCs were cultured in serum-free cell culture. Individual cultures were assigned to one of 4 treatment groups: Control, Hypoxia, folate supplementation (Folate) and folate deficiency (Folate-D). The latter 3 groups were exposed to hypoxia for 6 h on the 3rd day，then be collected on the 7th day．Cell proliferation was assessed by methyl thiazolyl tetrazolium assay. The expression levels of Notch1, Hes1/5, Mash1and Ngn1/2 mRNA were analyzed by real-time PCR. Results NSCs vitality in Folate group was higher than that in Hypoxia group. Proliferation of NSCs was lowest in Folate-D group (P<0.05). Notch1 and Hes1/5 mRNA expression levels in Folate group were higher than those in Hypoxia group (P<0.05). The expression levels of those three genes were lowest in Folate-D group (P<0.05). The expression levels of Ngn1/2 mRNA in Folate group were lower than those in Hypoxia group (P<0.05). There was no statistical significance in expression level of Mash1 mRNA in all groups (P>0.05). Conclusion Supplementation of NSCs with folate after hypoxia may stimulate cells proliferation and the mRNA expression levels of Notch1 and Hes1/5. [ACTA NUTRIMENTA SINICA, 2011,33(6)：567－570]

Key words： neural stem cell; folate; hypoxia; Notch signal pathway

脑梗死是神经系统常见病和多发病，已成为目前人类疾病的三大死亡原因之一。近年来，利用神经干细胞（neural stem cells，NSCs）移

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植治疗脑梗死取得了一定进展。NSCs的增殖

与分化受多种因素影响，包括细胞自身基因和外来信号的调控，了解缺氧损伤下NSCs的增殖分化调控机制有助于指导临床应用NSCs技术治疗脑梗死等疾病。前期研究中我们已发现叶酸能够

收稿日期 2011-07-03

基金项目 国家自然科学基金（No. 81072289）； 作者简介 刘欢（1980－），女，博士，讲师， E-mail：liuhuan@tijmu.edu.cn； 通讯作者：黄国伟，E-mail：guoweihuang@yahoo.com.cn

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