内源性大麻素及其代谢酶与孤独症谱系障碍严重程度的相关性

目的探讨内源性大麻素(endocannabinoid,eCB)及其代谢酶与孤独症谱系障碍(autism spectrum disorder,ASD)的关联,为ASD的病因及发病机制研究提供理论依据。方法采用病例对照的研究方法,收集2017年6月至2018年12月在哈尔滨医科大学儿童发育行为研究中心就诊和在省孤独症定点康复机构接受康复训练的58名ASD儿童作为ASD组。按照性别、年龄1∶1匹配的原则,在黑龙江省抽取58名正常发育儿童作为对照组。采集两组儿童空腹静脉血,采用液相色谱-质谱联用技术(LC-MS/MS)和实时荧光定量PCR(RT-qPCR)检测其外周血中内源性大麻素花生四烯酸乙醇胺(AEA)、花生四烯酸甘油(2-AG)、十六酰胺乙醇(PEA)、油酰乙醇胺(OEA)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)、酰胺水解酶(FAAH)、单酰基甘油脂酶(MAGL)和二酰基甘油脂酶(DAGL)的mRNA表达水平,采用Pearson相关分析eCB与ASD儿童症状严重程度之间的相关性。结果ASD儿童的AEA、OEA和PEA的水平[(10.10±2.60)nmol/L,(24.30±5.60)nmol/L,(15.92±2.28)nmol/L]均低于对照组儿童[(13.46±3.04)nmol/L,(27.85±6.89)nmol/L,(17.87±2.67)nmol/L,t=-6.612,-3.089,-4.579,均P<0.01];ASD儿童FAAH和DAGL mRNA的表达水平显著高于对照组,且差异有统计学意义(t=2.423,3.840,P<0.05),而NAPE-PLD和MAGL mRNA水平在两组间差异无统计学意义(t=0.024,0.885,均P>0.05);ASD组PEA水平与儿童孤独症行为量表(ABC)总分呈负相关(r=-0.288,P<0.05)。结论ASD儿童体内的eCB及其代谢酶可能存在代谢异常,且eCB水平与ASD的严重程度存在关联性。

Association of endocannabinoid and its metabolic enzymes with severity in children with autism spectrum disorder

Objective To explore the relationship between endocannabinoid(eCB)and its metabolic enzymes and severity of autism spectrum disorder(ASD),and to provide a theoretical basis for the study of the etiology and pathogenesis of ASD.Methods A case-control study was conducted to collect 58 ASD children who underwent rehabilitation training at the Children's Developmental Behavior Research Center of Harbin Medical University and the provincial autism rehabilitation facility from December 2017 to December 2018 as the ASD group.According to the principle of gender and age 1∶1 matching,58 normal children were selected as control group in Heilongjiang Province.Liquid chromatography-tandem mass spectrometry(LC-MS/MS)and real-time quantitative PCR(RT-qPCR)were used to detect eCB of ASD group and control group,including anandamide(AEA),2-arachidonoylglycerol(2-AG),palmitoylethanolamide(PEA),oleoylethanolamide(OEA)and its metabolic enzymes:n-acylphosphatidylethanolamine-specific phospholipase D(NAPE-PLD),fatty acid amide hydrolase(FAAH),monoacylglycerol lipase(MAGL)and diacylglycerol lipase(DAGL)mRNA expression levels.Pearson correlation was used to analyze the level of eCB and ASD children's severity.Results The levels of AEA,OEA and PEA in ASD children((10.10±2.6)nmol/L,(24.30±5.60)nmol/L,(15.92±2.28)nmol/L)were lower than those in the control group((13.46±3.04)nmol/L,(27.85±6.89)nmol/L,(17.87±2.67)nmol/L,t=-6.612,-3.99,-4.779,P<0.01).The expression levels of FAAH and DAGL mRNA in ASD children were significantly higher than those in the control group,and the difference was statistically significant(t=2.423,3.840,P<0.05),while NAPE-PLD and MAGL mRNA levels were not significantly different between the two groups(t=0.024,0.885,P>0.05).The level of PEA in the ASD group was negatively correlated with the total score of the autism behavior checklist(ABC)(r=-0.288,P<0.05).Conclusion There may be metabolic abnormalities in eCB and its metabolic enzymes in ASD children,and the level of eCB is related with the severity of ASD.