同一胎鼠肠和脑神经干细胞神经递质的表达

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摘要目的：观察同一胎鼠肠神经干细胞和脑神经干细胞的神经递质表达情况，探索神经干细胞移植治疗相关疾病的合适干细胞源。方法：从孕20 d的同一SD胚鼠中分别提取肠神经干细胞和脑神经干细胞，采用改良的神经干细胞培养液反复传代，取传代培养6～10 d后得到的2种不同来源的神经细胞球，通过琼脂糖预包埋免疫细胞化学检测法及流式细胞仪检测法测定神经干细胞4种神经递质的表达。结果：2种不同来源的神经干细胞的琼脂糖预包埋免疫细胞化学检测显示：肠神经干细胞神经元型一氧化氮合酶（nNOS）、胆碱乙酰转移酶（ChAT）与相应的对照组比较差异有统计学意义（P＜0.05）；脑神经干细胞nNOS、ChAT、多巴胺β羟化酶（DβH）与相应的对照组比较差异有统计学意义（P＜0.05）；2种不同来源神经干细胞间nNOS、ChAT比较差异有统计学意义（P＜0.05）。流式细胞仪检测提示：肠神经干细胞γ-氨基丁酸（GABA）、nNOS、ChAT、DβH阳性细胞合计率分别为1.59%、14.97%、5.43%、1.36%，其中nNOS、ChAT与相应的同型对照组比较差异有统计学意义（P＜0.05）；而脑神经干细胞GABA、nNOS、ChAT、DβH阳性细胞合计率分别为1.68%、6.14%、25.42%、9.30%，其中nNOS、ChAT、DβH与相应的同型对照组比较差异有统计学意义（P＜0.05）。结论：肠神经干细胞表达的神经递质主要为一氧化氮（NO）和乙酰胆碱（Ach），以NO为主；脑神经干细胞表达的神经递质为NO、Ach和去甲肾上腺素（NA），以Ach为主；2种神经干细胞均无明显表达GABA。

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	夏立广
	李仲荣
	刘森清
	王永飚
	朱利斌
	赵一鸣

关键词 ： , 神经干细胞；神经递质；表达；胎鼠

Abstract：Objective: To observe the expression of neurotransmitters in enterogenous neural stem cells (ENSCs) and central nervous system neural stem cells (CNS-NSCs) from the same fetal rats, and to explore a suitable stem cell source for neural stem cells transplantation. Methods: Rat ENSCs and CNS-NSCs were extracted from the same 20-day SD rat embryos followed by repeated subculture using improved neural stem cell medium. Two different kinds of nerve bulbs originated from 6-10 days culturing were chosen. The expressions of four transmitters on ENSCs and CNS-NSCs were analyzed by agarose pre-embedded immunocytochemistry assay and flow cytometry. Results: After analyzing data of imunocytochemistry assay, we found the expression of nNOS and ChAT on ENSCs; nNOS, ChAT and DβH on CNS-NSCs. Significant differences of nNOS and ChAT expression were observed between ENSCs and CNS-NSCs respectively (P<0.05). Flow cytometry analysis indicated GABA(+), nNOS(+), ChAT(+), DβH(+) cells averagely accounted for 1.59%, 14.97%, 5.43%, 1.36% in ENSCs respectively,within which nNOS and ChAT expression were significantly different from isotype groups (P<0.05). In CNS-NSCs, the pecentages were 1.68%, 6.14%, 25.42%, 9.30% on average, comparing with isotype groups, the differences of nNOS, ChAT, and DβH expression were all significant (P<0.05). Conclusion: ENSCs express neurotransmitters including NO and Ache, with NO of dominance. While CNS-NSCs express neurotransmitters such as NO, Ache and NE, especially Ache. There is almost no GABA expression in either ENSCs or CNS-NSCs.

Key words： NSCs neurotransmitter expression fetal rats

收稿日期: 2014-08-31

基金资助:浙江省自然科学基金资助项目( Y207272)；温州市科技局科技计划项目( Y20100134)。

通讯作者: 赵一鸣，副主任医师，Email：zym81835132@126.com。

作者简介: 夏立广（1984-），男，浙江温州人，住院医师，硕士。

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2014/V44/I11/792

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