七氟醚通过FAS-FASL通路致幼鼠学习记忆损伤的研究

Study on sevoflurane induced learning and memory impairment in young mice through FAS-FASL signaling Pathway

目的研究七氟醚可致幼鼠神经细胞凋亡增加并导致其学习记忆障碍的具体机制。方法将60只野生型幼鼠分成四组进行试验,对照组15只(野生型幼鼠+纯氧)、七氟醚组15只(野生型幼鼠+七氟醚)、FAS基因敲除组15只(敲除FAS基因幼鼠+七氟醚)、FASL基因敲除组15只(敲除FASL基因幼鼠+七氟醚),暴露结束后4 h,从四组幼鼠中各取5只处死,取海马组织,用原位末端标记法(TUNEL)计算海马处的凋亡细胞数量,用WB方法检测FAS蛋白表达量。剩余的幼鼠在第30~34天进行水迷宫试验检测其学习记忆能力。结果吸入七氟醚的幼鼠FAS蛋白表达明显升高,与敲除FAS基因的幼鼠、敲除FASL基因的幼鼠相比,吸入七氟醚的野生型幼鼠可以观察到较多的原位末端标记法阳性的海马细胞。水迷宫实验表明,与敲除FAS基因的幼鼠、敲除FASL基因的幼鼠相比,吸入七氟醚的野生型幼鼠在逃避潜伏期及通过目标象限的次数均较长。结论这些结果表明,七氟醚可通过FAS-FASL信号通路导致幼鼠神经细胞凋亡增加并导致学习记忆损伤。

Objective To study the mechnism of Sevoflurane can induce neuron apoptosis and cause learning and memory impairment in young mice. Methods There were four groups,control group（ Wild-type young mice ＋ pure oxygen）,sevoflurane group（ wild-type young mice ＋ sevoflurane）,FAS-knockout group（ FAS-knockout young mice ＋ sevoflurane）,FASL-knockout group（ FASL-knockout mice ＋ sevoflurane）. Four hours after the exposure,6 rats from each group were sacrificed,and the hippocampus was removed. We used TUNEL staining to count the apoptotic cells in hippocampus,and Western Blotting to study the expression of FAS protein. The remain young rats were tested for their learning and memory in the water maze on days 30-34. Results We found that sevoflurane significantly increased expression of FAS protein in wild type mice. Compared to FAS-and FASL-knockout mice treated with sevoflurane,sevoflurane-treated wild-type mice had showed more TUNEL-positive hippocampal cells. Morris water maze showed that compared with FAS-and FASL-knockout mice treated with sevoflurane,sevoflurane treatment of wild type mice significantly prolonged escape latency and reduced platform crossing times. Conclusion These data suggested that sevoflurane induced neuron apoptosis and learning and memory impairment in young mice through FAS-FASL signaling pathway.