中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2018, Vol. 13 ›› Issue (9): 1575-1584.doi: 10.4103/1673-5374.237121

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

孕中期高浓度大鼠暴露七氟醚可诱导子代神经干细胞凋亡

  

  • 收稿日期:2018-06-25 出版日期:2018-09-15 发布日期:2018-09-15
  • 基金资助:

    国家自然科学基金(81671311、81503273)、中国辽宁省科学技术基金(2015020467)、中国医科大学盛京医院优秀科学基金(201708)

High-concentration sevoflurane exposure in mid-gestation induces apoptosis of neural stem cells in rat offspring

Yuan Wang1, Shao-Wei Yin2, Nan Zhang3, Ping Zhao1   

  1. 1 Department of Anesthesiology, Shengjing Hospital of China Medical University, Shenyang, Liaoning Province, China
    2 Department of Obstetrics, Shengjing Hospital of China Medical University, Shenyang, Liaoning Province, China
    3 Department of Neuroendocrine Pharmacology, School of Pharmacy, China Medical University, Shenyang, Liaoning Province, China
  • Received:2018-06-25 Online:2018-09-15 Published:2018-09-15
  • Contact: Ping Zhao, M.D., Ph.D.,zhaop@sj-hospital.org
  • Supported by:

    This work was supported by the National Natural Science Foundation of China, No. 81671311 (to PZ), No. 81503273 (to NZ); the Science and Technology Foundation of Liaoning Province of China, No. 2015020467 (to PZ); the Outstanding Scientific Fund of Shengjing Hospital of China Medical University, No. 201708.

PDF

174

摘要：

七氟醚是孕期最常用的吸入麻醉药物。子代脑组织神经干细胞的存活状态直接影响着子代的脑发育，然而，孕中期应用七氟醚是否影响子代神经干细胞存活目前尚不清楚。因此，我们观察了孕中期暴露于七氟醚的子代是否会引起神经干细胞凋亡，是否会在生后出现行为学异常，并探讨了可能存在的机制。实验设计给予孕14 d大鼠2%和3.5%七氟醚麻醉2 h，在生后28 d取子代鼠行水迷宫测试，免疫荧光染色检测麻醉后6，24，48 h胎鼠脑组织或生后0，14，28 d子代海马凋亡神经干细胞数量变化，通过Western Blot方法麻醉后6，24，48 h胎鼠脑组织或生后0，14，28 d子代海马Nestin、SOX-2蛋白水平变化，以Western Blot方法检测麻醉6 h后胎鼠脑组织血管内皮生长因子以及PI3K/AKT通路蛋白水平。发现大鼠孕中期接受高浓度(3.5%)七氟醚麻醉引起子代鼠在水迷宫测试中逃避潜伏期及上台前路程延长，穿越平台次数及目标象限活动时间减少；同时胎鼠（麻醉后6，24，48 h）脑组织或生后子代（生后0，14，28 d）海马Nestin/TUNEL阳性细胞百分比增加，Nestin、SOX-2蛋白水平降低，麻醉6 h后胎鼠脑组织血管内皮生长因子及PI3K和p-AKT蛋白表达降低；而接受低浓度七氟醚(2%)麻醉的胎鼠和子代却未出现上述变化。上述数据说明，孕中期大鼠应用高浓度七氟醚(3.5%)造成子代发育期脑损伤与吸入浓度有关，高浓度七氟醚可能通过降低血管内皮生长因子，PI3K及p-AKT蛋白表达诱导子代海马神经干细胞凋亡，进而引起学习记忆功能异常。

orcid:0000-0001-6392-5391(Ping Zhao)

关键词: 七氟醚, 神经干细胞, 凋亡, 学习记忆, 血管内皮生长因子, PI3K, p-AKT, 麻醉, 学习, 记忆, 发育神经生物学, 神经再生

Abstract:

Sevoflurane is the most commonly used volatile anesthetic during pregnancy. The viability of neural stem cells directly affects the development of the brain. However, it is unknown whether the use of sevoflurane during the second trimester affects the survival of fetal neural stem cells. Therefore, in this study, we investigated whether exposure to sevoflurane in mid-gestation induces apoptosis of neural stem cells and behavioral abnormalities. On gestational day 14, pregnant rats were anesthetized with 2% or 3.5% sevoflurane for 2 hours.The offspring were weaned at 28 days and subjected to the Morris water maze test. The brains were harvested to examine neural stem cell apoptosis by immunofluorescence and to measure Nestin and SOX-2 levels by western blot assay at 6, 24 and 48 hours after anesthesia as well as on postnatal day (P) 0, 14 and 28. Vascular endothelial growth factor (VEGF) and phosphoinositide 3-kinase (PI3K)/AKT pathway protein levels in fetal brain at 6 hours after anesthesia were assessed by western blot assay. Exposure to high-concentration (3.5%) sevoflurane during mid-gestation increased escape latency and path length to the platform, and it reduced the average duration spent in the target quadrant and platform crossing times. At 6, 24 and 48 hours after anesthesia and at P0, P14 and P28, the percentage of Nestin/terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL)-positive cells was increased, but Nestin and SOX-2 protein levels were decreased in the hippocampus of the offspring. At 6 hours after anesthesia, VEGF, PI3K and phospho-AKT (p-AKT) levels were decreased in the fetal brain. These changes were not observed in animals given low-concentration (2%) sevoflurane exposure. Together, our findings indicate that exposure to a high concentration of sevoflurane (3.5%) in mid-gestation decreases VEGF, PI3K and p-AKT protein levels and induces neural stem cell apoptosis, thereby causing learning and memory dysfunction in the offspring.

Key words: nerve regeneration, sevoflurane, neural stem cells, apoptosis, vascular endothelial growth factor, PI3K, p-AKT, anesthesia, learning, memory, developmental neurobiology, neural regeneration