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。

目的 探讨花生四烯酸氨基乙醇(anandamide,AEA)对人成牙本质细胞(HODs)样细胞中基质金属蛋白酶(MMP)-9表达的影响.方法 体外培养HODs样细胞,免疫荧光进行形态和功能验证,四甲基偶氮唑盐(MTT)法研究AEA对HODs样细胞活性影响,实时荧光定量聚合酶链反应(RT-qPCR)和Western blotting检测AEA处理后MMP-9基因和蛋白表达水平及AEA的作用受体大麻素受体1,2和TRPV1的表达,并检测MAPK信号通路的调控作用.结果 大麻素受体1,2和TRPV1在HODs样细胞中表达,不同浓度AEA和不同处理时间可刺激MMP-9基因和蛋白表达水平提高.抑制大麻素受体1,2和TRPV1后AEA诱导的MMP-9表达水平下降,其中大麻素受体1和TRPV1作用较强.JNK信号通路在AEA对MMP-9的表达调节中起主导作用.结论 AEA主要通过大麻素受体1和TRPV1调控HODs样细胞中MMP-9的表达,JNK信号通路是此过程的主要调节通路.

Cannabinoids are lipophilic substances derived from Cannabis sativa that can exert a variety of effects in the human body.They have been studied in cellular and animal models as well as in human clinical trials for their therapeutic benefits in several human diseases.Some of these include central nervous system(CNS)diseases and dysfunctions such as forms of epilepsy,multiple sclerosis,Parkinson's disease,pain and neuropsychiatric disorders.In addition,the endogenously produced cannabinoid lipids,endocannabinoids,are critical for normal CNS function,and if controlled or modified,may represent an additional therapeutic avenue for CNS diseases.This review discusses in vitro cellular,ex vivo tissue and in vivo animal model studies on cannabinoids and their utility as therapeutics in multiple CNS pathologies.In addition,the review provides an overview on the use of cannabinoids in human clinical trials for a variety of CNS diseases.Cannabinoids and endocannabinoids hold promise for use as disease modifiers and therapeutic agents for the prevention or treatment of neurodegenerative diseases and neurological disorders.

CYP4X1是CYP4新的亚家族成员,也是重要的Orphans CYPs成员之一.其核苷酸序列同源性分析显示CYP4X1结构在跨物种间是高度保守的,这预示着它具有重要的生物学功能.CYP4X1广泛存在于人体各组织中,尤其选择性在脑内高表达,提示其可能在神经血管功能中扮演关键作用.CYP4X1的表达存在昼夜节律调节、明显的性别差异和年龄差异以及外源物可诱导其表达.CYP4X1重要的生物学功能之一是代谢内源性大麻素Anandamide生成唯一的单加成产物14,15-EET-EA以及参与脂肪的代谢.EET-EAs可能发挥着EETs相似的生物学功能.此外,CYP4X1与肿瘤的分级有关,其可能成为肿瘤治疗的潜在药物靶标.

Anandamide is a lipid mediator that acts as an endogenous ligand of CB1 receptors.These receptors are also the primary molecular target responsible for the pharmacological effects of A9-tetrahydrocannabinol,the psychoactive ingredient in Cannabis sativa.Several studies demonstrate that anandamide exerts an overall modulatory effect on the brain reward circuitry.Several reports suggest its involvement in the addiction-producing actions of other abused drugs,and it can also act as a behavioral reinforcer in animal models of drug abuse.Importantly,all these effects of anandamide appear to be potentiated by pharmacological inhibition of its metabolic degradation.Enhanced brain levels of anandamide after treatment with inhibitors of fatty acid amide hydrolase,the main enzyme responsible for its degradation,seem to affect the rewarding and reinforcing actions of many drugs of abuse.In this review,we will provide an overview from a preclinical perspective of the current state of knowledge regarding the behavioral pharmacology of anandamide,with a particular emphasis on its motivational/reinforcing properties.We will also discuss how modulation of anandamide levels through inhibition of enzymatic metabolic pathways could provide a basis for developing new pharmaco-therapeutic tools for the treatment of substance use disorders.

Cannabis sativa,is also popularly known as marijuana,has been cultivated and used for recreational and medicinal purposes for many centuries.The main psychoactive content in cannabis is △9-tetrahydrocannabinol (THC).In addition to plant cannabis sativa,there are two classes of cannabinoids—the synthetic cannabinoids (e.g.,WIN55212-2) and the endogenous cannabinoids (eCB),anandamide (ANA) and 2-arachidonoylglycerol (2-AG).The biological effects of cannabinoids are mainly mediated by two members of the G-protein-coupled receptor family,cannabinoid receptors 1 (CB1R) and 2 (CB2R).The endocannabinoids,cannabinoid receptors,and the enzymes/proteins responsible for their biosynthesis,degradation,and re-updating constitute the endocannabinoid system.In recent decades,the endocannabinoid system has attracted considerable attention as a potential therapeutic target in numerous physiological conditions,such as in energy balance,appetite stimulation,blood pressure,pain modulation,embryogenesis,nausea and vomiting control,memory,learning and immune response,as well as in pathological conditions such as Parkinson's disease,Huntington's disease,Alzheimer's disease,and multiple sclerosis.

Nervous system emerges from complex signaling interactions where extrinsic (neurotransmitters and trophic factors, among others) and intrinsic factors (transcription factors) interplay in the developing tissue to control gene activity promoting chron-ic changes in cell genesis, migration, differentiation and death. The retinal microenvironment is regulated by a broad variety of chemicals, including endocannabinoids and nucleotides that mod-ulate embryonic progenitor-neuron-Müller glia signaling in very early developing or pathophysiological conditions. Accumulated evidence demonstrate the presence of a functional cannabinoid system in this tissue, with many retina cell types expressing canna-binoid CB1 and/or CB2 receptors, the two main ligands N-arachi-donoylethanolamide (anandamide) and 2-arachidonoylglycerol (2-AG) and enzymes that generate N-acyl phosphatidylethanolamine phospholipase, and diacylglycerol lipase (DAGL) and degrade fatty acid amide hydrolase, monoacylglycerol lipase (MAGL) and cyclooxygenase-2 endocannabinoids (Kokona et al., 2016). As in other areas of the central nervous system, cannabinoids seem to regulate neurotransmission in the retina by inhibiting the release of transmitters such as dopamine, norepinephrine, γ-aminobutyric acid and glutamate. They also play an important role in retinal cir-cuitry and in scotopic vision by modulating Ca2+ and K+ channels in bipolar cells and photoreceptors. Moreover, under pathological conditions, cannabinoids seem to induce neuroprotection in this tissue (Kokona et al., 2016). Albeit these protective effects, howev-er, recent data are implicating cannabinoid receptors in cell death in the retina, both in the early developing (Freitas et al., 2019) and diseased tissue (Matias et al., 2006; El-Remessy et al., 2011; Chen et al., 2018).

内源性大麻素系统是包括了内源性大麻素、大麻素受体(CB)以及相关酶的一个复杂的生理系统.近年来内源性大麻素系统与骨代谢的关系被一步步证实.大量研究发现CB-1受体及CB-2受体存在于骨细胞中,并在骨代谢的过程中起到不同的作用.其余受体包括TPRV1、GPR55、GPR119、TPRV4等,亦参与了骨代谢的过程.以anandamide和2-arachidonoylglycerol为代表的内源性大麻素以及其激动剂和拮抗剂可以通过大麻素受体的介导影响成骨细胞与破骨细胞的增殖分化及骨重塑.可见内源性大麻素系统可作为骨再生治疗的新靶点,然而目前尚无足够数量的相关临床数据,因此大麻素与骨代谢的关系值得进一步研究.

Rosiglitazone(RSG)is a synthetic agonist of peroxisome proliferator-activated receptor-y(PPARy),which plays a central role in the regulation of metabolism.Meta-analyses have suggested that RSG is associated with increased cardiovascular risk.However,the mechanisms underlying such adverse cardiac effects are still poorly understood.Here,we found that activation of PPARy by RSG stimulated the endocannabinoid system(ECS),a membrane lipid signaling system,which induced cardiac hypertrophy.In neonatal rat cardiomyocytes,RSG increased the level of anandamide(AEA);upregulated the expression of N-acyl phosphatidylethanolamine phospholipase D(NapePLD),a key enzyme for AEA synthesis;and downregulated the expression of fatty acid amide hydrolase(FAAH),the enzyme responsible for the degradation of AEA.Importantly,PPARy activation increased the expression of cannabinoid receptor type 1(CB1)through an identified binding site for PPARy in the CB1 promoter region.Moreover,both the in vitro and in vivo results showed that inhibition of the ECS by rimonabant,an antagonist of CB1,attenuated RSG-induced cardiac hypertrophy,as indicated by decreased expression of cardiac hypertrophy markers(ANP and BNP),deactivation of the mTOR pathway,and decreased cardiomyocyte size.Thus,these results demonstrated that the ECS functions as a novel target of PPARy and that the AEA/CB1/mTOR axis mediates RSG-induced cardiac remodeling.