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
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References
[1] US APAD-TFAV. Diagnostic and statistical manual of mental disorders: DSM-5TM[M].5th ed.Codas,2013.
[2] Elsabbagh M,Divan G,Koh YJ,et al. Global prevalence of autism and other pervasive developmental disorders[J]. Autism Res,2012,5(3):160-179.
[3] Baio J,Wiggins L,Christensen DL,et al. Prevalence of autism spectrum disorder among children aged 8 years-autism and developmental disabilities monitoring network, 11 sites, United States, 2014[J]. MMWR Surveill Summ,2018,67(6):1-23.
[4] Servadio M,Melancia F,Manduca A,et al. Targeting anandamide metabolism rescues core and associated autistic-like symptoms in rats prenatally exposed to valproic acid[J]. Transl Psychiatry,2016,6(9):e902.
[5] Nguyen QA,Horn ME,Nicoll RA. Distinct roles for extracellular and intracellular domains in neuroligin function at inhibitory synapses[J]. Elife,2016,5:e19236.
[6] Hamzawy MA,El-Ghandour YB,Abdel-Aziem SH,et al. Leptin and camel milk abate oxidative stress status, genotoxicity induced in valproic acid rat model of autism[J]. Int J Immunopathol Pharmacol,2018,32:2058738418785514.
[7] Wu H,Zhang Q,Gao J,et al. Modulation of sphingosine 1-phosphate (S1P) attenuates spatial learning and memory impairments in the valproic acid rat model of autism[J]. Psychopharmacology (Berl),2018,235(3):873-886.
[8] Aran A,Eylon M,Harel M,et al. Lower circulating endocannabinoid levels in children with autism spectrum disorder[J]. Mol Autism,2019,10:2.
[9] Karhson DS,Krasinska KM,Dallaire JA,et al. Plasma anandamide concentrations are lower in children with autism spectrum disorder[J]. Mol Autism,2018,9:18.
[10] Siniscalco D,Sapone A,Giordano C,et al. Cannabinoid receptor type 2, but not type 1, is up-regulated in peripheral blood mononuclear cells of children affected by autistic disorders[J]. J Autism Dev Disord,2013,43(11):2686-2695.
[11] Siniscalco D,Bradstreet JJ,Cirillo A,et al. The in vitro GcMAF effects on endocannabinoid system transcriptionomics, receptor formation, and cell activity of autism-derived macrophages[J]. J Neuroinflammation,2014,11:78.
[12] Kerr DM,Gilmartin A,Roche M. Pharmacological inhibition of fatty acid amide hydrolase attenuates social behavioural deficits in male rats prenatally exposed to valproic acid[J]. Pharmacol Res,2016,113(Pt A):228-235.
[13] Zamberletti E,Gabaglio M,Woolley-Roberts M,et al. Cannabidivarin treatment ameliorates autism-like behaviors and restores hippocampal endocannabinoid system and glia alterations induced by prenatal valproic acid exposure in rats[J]. Front Cell Neurosci,2019,13:367.
[14] Neumeister A,Normandin MD,Pietrzak RH,et al. Elevated brain cannabinoid CB1 receptor availability in post-traumatic stress disorder: a positron emission tomography study[J]. Mol Psychiatry,2013,18(9):1034-1040.
[15] Melancia F,Schiavi S,Servadio M,et al. Sex-specific autistic endophenotypes induced by prenatal exposure to valproic acid involve anandamide signalling[J]. Br J Pharmacol,2018,175(18):3699-3712.
[16] Zou M,Li D,Li L,et al. Role of the endocannabinoid system in neurological disorders[J]. Int J Dev Neurosci,2019,76:95-102.
[17] Kerr DM,Downey L,Conboy M,et al. Alterations in the endocannabinoid system in the rat valproic acid model of autism[J]. Behav Brain Res,2013,249:124-132.
[18] Maier T,Guell M,Serrano L. Correlation of mRNA and protein in complex biological samples[J]. FEBS Lett,2009,583(24):3966-3973.