中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (1): 118-122.doi: 10.3969/j.issn.2095-4344.2016.01.021

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

异氟醚对新生大鼠海马神经干细胞增殖及分化的影响

闵 娜,胡强夫,李晓培,聂晓红,杨利利   

  1. 郑州大学第五附属医院麻醉科,河南省郑州市 450052
  • 收稿日期:2015-11-23 出版日期:2016-01-01 发布日期:2016-01-01
  • 作者简介:闵娜 ,女,1979年生,广东省信宜市人,汉族,2004年牡丹江医学院毕业,主治医师,主要从事心血管麻醉方面的研究。

Isoflurane effects on the proliferation and differentiation of neural stem cells in the hippocampus of neonatal rats

Min Na, Hu Qiang-fu, Li Xiao-pei, Nie Xiao-hong, Yang Li-li   

  1. Department of Anesthesiology, Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China
  • Received:2015-11-23 Online:2016-01-01 Published:2016-01-01
  • About author:Min Na, Attending physician, Department of Anesthesiology, Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan Province, China

摘要:

文章快速阅读:

文题释义:

神经干细胞:存在于神经发生过程以及成年哺乳类动物中枢神经系统的神经干细胞是一种多能性细胞,具有自我更新以及分化为神经元、星形胶质细胞以及少突胶质细胞能力。从哺乳动物胚胎及脑组织中可以分离培养出神经干细胞,呈聚集式悬浮生长,形成典型的神经球形态,可表达其特异性标志物巢蛋白、CD133。
吸入麻醉:为挥发性麻醉药或麻醉气体经呼吸系统吸入,抑制中枢神经系统而产生全身麻醉的麻醉方法。吸入麻醉由于所用麻醉药在药理学性质上存在差别,其临床麻醉表现如诱导快慢、麻醉强度、苏醒快慢、对循环和呼吸的影响等也不相同,常用的吸入麻醉药物有乙醚、氟烷及七氟烷等。

 

背景:异氟醚麻醉药物会对神经系统产生一定的影响,研究表明其可能是通过影响神经干细胞功能或形态而引起神经功能障碍。
目的:探讨异氟醚对大鼠海马神经干细胞增殖和分化的影响。
方法:培养新生7 d SD大鼠海马神经干细胞并进行诱导分化,取传至第3代细胞,将其分为2组:异氟醚组通入2.8%异氟醚和体积分数为5%CO2,体积分数为95%O2的混合气体;对照组仅通入体积分数为5%CO2,体积分数为95%O2的混合气体。干预6 h后,再常规培养2 h。抗BrdU单克隆抗体进行免疫荧光染色检测细胞增殖情况,Western blot法测定神经元标志物β3-微管蛋白和胶质细胞标志物GFAP蛋白的表达。
结果与结论:与对照组相比,异氟醚组BrdU阳性细胞数目明显减少,说明异氟烷抑制了神经干细胞增殖;与对照组相比,异氟醚组GFAP表达水平上调,β3-微管蛋白表达水平无明显改变,说明异氟烷可促进神经干细胞更多地向星形胶质细胞分化。 

ORCID: 0000-0001-8738-9327(闵娜)

关键词: 干细胞, 培养, 异氟醚, 吸入麻醉, 大鼠, 新生, 海马, 神经干细胞, 细胞增殖, 神经元分化

Abstract:

BACKGROUND: Isoflurane is an anesthesia drug that has a certain effect on the nervous system. It possibly causes neurologic disorders through impacting nerve stem cell function or morphology.
OBJECTIVE: To investigate the effects of isoflurane on the proliferation and differentiation of neural stem cells in the hippocampus of rats.
METHODS: Neural stem cells from the hippocampus of neonatal Sprague-Dawley rats, aged 7 days, were induced and differentiated. Passage 3 cells were obtained and divided into two groups: isoflurane group (a mixture gas of 2.8% isoflurane, 5% CO2 and 95% O2) and control group (a mixture of 5% CO2 and 95% O2). After  intervention of 6 hours followed by 2 hours of routine culture, anti-BrdU monoclonal antibody immunofluorescent staining was used to detect cell proliferation, and western blot assay to detect the expression of β3-tubulin and glial fibrillary acidic protein.
RESULTS AND CONCLUSION: Compared with the control group, the number of BrdU positive cells in the isoflurane group reduced significantly, indicating that isoflurane inhibits the proliferation of neural stem cells. Compared with the control group, the expression of glial fibrillary acidic protein in the isoflurane group up-regulated, but the expression of β3-tubulin had no changes, indicating isoflurane promotes the differentiation of neural stem cells into astrocytes.
 

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