journal1 ›› 2020, Vol. 28 ›› Issue (4): 411-415.DOI: 10.11852/zgetbjzz2020-0010

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Study on expression of N-acylethanolamine-related receptors and metabolic enzymes in hippocampus of autism spectrum disorder model rats

LI Ling, LI De-xin, LIU Yu, XIE Shu, ZOU Ming-yang, SUN Cai-hong, WU Li-jie   

  1. Department of Children′s and Adolescent Health, Public Health College, Harbin Medical University, Harbin, Heilongjiang 150081, China
  • Received:2020-01-03 Online:2020-04-10 Published:2020-04-10
  • Contact: WU Li-jie, E-mail:wulijiehyd@126.com

孤独症模型鼠海马N-酰基乙醇胺相关受体及代谢酶的表达研究

李玲, 李德欣, 刘瑜, 谢姝, 邹明扬, 孙彩虹, 武丽杰   

  1. 哈尔滨医科大学公共卫生学院儿少卫生与妇幼保健学教研室,黑龙江 哈尔滨 150081
  • 通讯作者: 武丽杰,E-mail:wulijiehyd@126.com
  • 作者简介:李玲(1993-),女,山东人,硕士在读,主要研究方向为儿童发育障碍及行为问题。
  • 基金资助:
    国家自然科学基金项目(81773458);国家自然科学基金项目(81903348)

Abstract: Objective To test the expression of N-acylethanolamines (NAEs) related receptors and the metabolic enzymes in the hippocampus of autism spectrum disorder (ASD) model rats, and to explore the correlation between NAEs signals and ASD, so as to provide new insight for the etiological study of ASD. Methods ASD model rats induced by sodium valproate (VPA) were studied for growth, development and behavioral tests. Western Blot and real-time quantitative PCR (RT-PCR) were used to detect the protein and mRNA expression of NAEs-related receptors, namely endocannabinoid receptor (CB1R and CB2R), and their metabolic enzymes, namely N-acylphosphatidylethanolamine-specific phospholipase D (NAPE-PLD) and fatty acid amide hydrolase (FAAH). Results The results of self-grooming experiment, three-box experiment and water maze experiment showed that VPA rats had repetitive behavior patterns, social interaction dysfunction, and learning and memory dysfunction. Western Blot results revealed there were significant differences between VPA and control rats on CB1R and FAAH protein expressions in hippocampus (t=3.033, 2.865, P<0.05), however the difference on CB2R and NAPE-PLD was not found (t=1.034, 0.290, P>0.05). The significant differences in mRNA expression levels of CB1R, CB2R, FAAH and NAPE-PLD were not found in the hippocampus between two groups (t=2.239, 0.747, 0.748, 0.012, P>0.05). Conclusion The protein expression levels of NAEs-related receptor and metabolic enzyme in hippocampus of ASD model rats are up-regulated, suggesting that low NAEs signal may be associated with the pathogenesis of ASD.

Key words: autism spectrum disorder, valiproic acid, N-acylethanolamines, protein expression, mRNA expression

摘要: 目的 检测N-酰基乙醇胺(NAEs)相关受体及其代谢酶在孤独症谱系障碍(ASD)模型鼠海马中的表达,分析NAEs信号与ASD的关联性,为ASD的病因研究提供新的思路。方法 以丙戊酸钠(VPA)诱导的ASD模型鼠为研究对象,进行生长发育和行为学检测;采用Western Blot和RT-PCR检测其海马组织中NAEs相关受体,即内源性大麻素受体(CB1R和CB2R)及其代谢酶花生四烯酸磷脂酰乙醇胺特异性磷脂酶D(NAPE-PLD)和酰胺水解酶(FAAH)的蛋白及mRNA表达水平。结果 自梳实验、三箱实验及水迷宫实验结果显示:与对照组鼠相比,VPA鼠存在重复的行为模式、社会交互能力障碍及学习、记忆能力障碍;Western Blot结果显示:与对照组鼠相比,VPA鼠海马CB1R与FAAH蛋白表达上调(t=3.033、2.865,P<0.05),CB2R以及NAPE-PLD蛋白表达在两组大鼠间差异无统计学意义(t=1.034、0.290,P>0.05),且RT-PCR结果显示:CB1R、CB2R、FAAH和NAPE-PLD的mRNA表达水平在两组大鼠海马中差异均无统计学意义(t=2.239、0.747、0.748、0.012,P>0.05)。结论 ASD模型鼠海马中NAEs受体及水解酶蛋白表达上调,提示低NAEs信号可能与ASD发病存在关联。

关键词: 孤独症谱系障碍, 丙戊酸钠, N-酰基乙醇胺, 蛋白表达, mRNA表达

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