Search Results (959)

Background: Cannabis contains Δ⁹-tetrahydrocannabinol (THC) together with numerous other constituents, whereas HU210 (HU) is a synthetic full agonist at cannabinoid receptors. We applied integrated metabolomics and proteomics to compare Cannabis extract (CS) and HU in GT1-7 neuronal cells. Methods: Cytotoxicity was evaluated using a lactate dehydrogenase assay. Metabolites and proteins were extracted sequentially from single-cell pellets and analyzed using gas chromatography–tandem mass spectrometry and nano-liquid chromatography–quadrupole–Orbitrap mass spectrometry, respectively. Cannabinoids in the exposure medium were quantified using liquid chromatography–tandem mass spectrometry. Results: The lowest CS and HU concentrations that were cytotoxic at 24 h and non-cytotoxic at 3 h were 2000 µg/mL and 20 µM, respectively; CS delivered 166 nM THC, 800 nM tetrahydrocannabinolic acid, and 11.3 nM cannabidiol. After 3 h, HU altered both the metabolome and the proteome, whereas CS altered the metabolome with minimal proteome change (8 of 2720 proteins). Six pathways differed, including the citrate cycle and pentose phosphate pathway. Relative to both the control and CS, HU decreased citrate; increased 2-ketoglutarate, fumarate, malate, gluconate, ribose-5-phosphate and ribose; and increased phosphoenolpyruvate carboxykinase 2, isocitrate dehydrogenase 3 subunit α, 2-oxoglutarate dehydrogenase complex component E1, succinate dehydrogenase flavoprotein subunit, malate dehydrogenase 1, 6-phosphogluconolactonase, 6-phosphogluconate dehydrogenase and phosphoglucomutase-1. CS showed the opposite metabolite changes without corresponding enzyme changes. Conclusions: At a pre-lethal time point, CS and HU produced metabolic changes in opposite directions—broad suppression versus coordinated mobilisation—despite comparable cytotoxicity at 24 h. Full article

Microglia are key regulators of central nervous system homeostasis and neuroinflammation, and cannabinoid type 2 receptors (CB₂Rs) have emerged as important modulators of microglial function. Although the pharmacology of CB₂Rs has been extensively characterised in heterologous expression systems and activated microglia, comparatively little is known about the behaviour of CB₂R ligands in non-stimulated microglia. The present study therefore aimed to characterise the pharmacological properties of a panel of CB₂R ligands in non-stimulated BV-2 microglial cells and to determine whether constitutive receptor activity and ligand-dependent signalling bias could be detected under basal conditions. Classical CB₂R agonists (CP 55,940, WIN 55,212-2, JWH 133 and JWH 015), putative protean agonists ((R)-AM 1241 and GW 405833), and inverse agonists (SR 144528, AM 630 and JTE 907) were evaluated using [³⁵S]GTPγS binding and forskolin-stimulated cAMP assays. In the [³⁵S]GTPγS assay, WIN 55,212-2 displayed the highest intrinsic activity, whereas (R)-AM 1241 and GW 405833 behaved as partial agonists. Inverse agonists reduced basal signalling, indicating constitutive CB₂R activity in resting BV-2 cells. In contrast, cAMP measurements revealed greater signal amplification, with (R)-AM 1241 and GW 405833 exhibiting full agonist behaviour and SR 144528 producing pronounced inverse agonism. Marked differences in ligand efficacy and rank order between assays highlighted the influence of downstream signalling mechanisms and ligand-dependent signalling bias. Notably, GW 405833 displayed a strongly biased signalling profile, whereas SR 144528 consistently exhibited the greatest inverse agonist activity. These findings demonstrate that CB₂Rs are functionally active in non-stimulated microglia and that constitutive receptor activity and signalling bias contribute significantly to their pharmacological profile under basal conditions. By focusing on non-stimulated microglia, this study provides new insights into CB₂R signalling in a homeostatic cellular environment and establishes a framework for understanding how CB₂R pharmacology may be altered during neuroinflammatory and neurodegenerative disease states. Full article

Endocannabinoid receptors of the CB1 subtype are the most abundant G-protein-coupled receptors in the central nervous system, where they strongly regulate neurotransmitter release. The effects of the activation of these receptors by exogenously applied agonists have also been described in the peripheral nervous system, particularly at neuromuscular junctions (NMJs). However, the physiological role of these receptors at NMJs has not been demonstrated. We have shown that blockade of CB1 receptors at the NMJs of newborn or young mice increases the quantal content of end-plate potentials, as well as their decay time constant. Neither effect of CB1 receptor blockade is observed if postsynaptic muscle acetylcholine receptors are partially blocked. Thus, CB1 receptors may be involved in maintaining synaptic homeostasis. Importantly, the effect of CB1 receptor blockade on quantal content is potentiated by blockade of the enzyme butyrylcholinesterase. Therefore, butyrylcholinesterase may be considered a component of the extracellular degradation system for endocannabinoids. Full article

Cannabis use has increased substantially in the United States, driven by broader legalization, decriminalization, and expanding medical and recreational availability. For facial plastic surgeons, the clinical implications remain difficult to define because “cannabis use” encompasses heterogeneous products and routes, including smoked flower, vaping, concentrates, edibles, pharmaceutical cannabinoids, topical cannabidiol (CBD), and frequent co-use with tobacco or nicotine. Current evidence suggests that systemic cannabis use, particularly inhaled or heavy perioperative use, may be associated with increased surgical complications in selected populations; however, existing studies are limited by retrospective design, inconsistent exposure definitions, inadequate dose and route characterization, and confounding by tobacco use and comorbidities. Cannabinoids exert biologic effects through the endocannabinoid system, particularly CB1 and CB2 receptors, which are expressed in the central nervous system, immune cells, vasculature, and skin. These pathways influence inflammation, keratinocyte proliferation, fibroblast activity, angiogenesis, immune surveillance, pain signaling, and tissue remodeling. The net effect of cannabinoid exposure on wound healing is likely context dependent, varying based on receptor expression, wound-healing phase, route of administration, cannabinoid composition, local tissue environment, and patient-specific risk factors. Preclinical and early dermatologic literature suggests potential therapeutic roles for topical cannabinoids, especially CBD, in modulating inflammation and epithelial repair. In contrast, systemic perioperative cannabis use has been associated in several surgical cohorts with infection, delayed healing, hematoma, nonunion, and reoperation. Evidence specific to facial plastic surgery remains sparse. The most directly relevant study evaluated cannabis and tobacco use in patients undergoing operative mandibular fracture repair. Cannabis-only use was not associated with increased complications, although the cohort was small; concurrent cannabis and tobacco use was associated with higher rates of surgical site infection, facial nonunion, abscess, debridement, and malocclusion. To date, no published studies address cannabis-associated outcomes in rhinoplasty, rhytidectomy, blepharoplasty, browlift, or facial rejuvenation. This review summarizes the biologic rationale, available surgical evidence, and clinical considerations for incorporating cannabis use into individualized perioperative risk assessment in facial plastic surgery. Full article

Substance use disorders (SUDs) remain a major public health concern and contribute substantially to compromised quality of life, mortality, and healthcare burden. In the United States alone, millions of individuals are affected by alcohol use disorder (AUD), tobacco use disorder (TUD), and opioid use disorder (OUD), with many cases complicated by co-existing anxiety and stress-related disorders. Piper methysticum G. Forst (kava), a traditional South Pacific plant preparation, has gained attention for its anxiolytic, sedative, and sleep-promoting properties. Its pharmacological effects are primarily attributed to a set of lipophilic compounds known as kavalactones, which have been reported to modulate GABA_A receptor activity, dopaminergic and adrenergic signaling pathways, monoamine oxidase-B activity, cannabinoid receptor type 1 activity, and voltage-gated ion channels. Peer-reviewed literature was identified through searches of PubMed, NIH resources, and other scientific databases using terms related to kava, kavalactones, addiction, anxiety, stress, insomnia, and SUDs. Both clinical and preclinical studies were reviewed, including investigations of neurotransmitter systems and addiction-related signaling pathways. The current literature suggests that the strongest rationale for kava use exists in AUD, where anxiety and stress are established contributors to relapse. Evidence supporting kava use in TUD and OUD is largely theoretical, while concerns regarding hepatotoxicity, cytochrome P450 interactions, product variability, and additive risk remain important barriers to its clinical application. In summary, current evidence does not support kava as a replacement for established therapies, while its unique pharmacological profile warrants further investigation as a potential adjunctive treatment for withdrawal and relapse in SUDs. Full article

In 1994, leptin was identified as a hormone that signals the brain to reduce food intake and increase energy expenditure. Leptin was initially considered a breakthrough therapy for obesity. This was largely based on animal studies, showing that leptin reversed obesity in individuals with congenital leptin deficiency and normal leptin receptor sensitivity. However, due to leptin resistance, leptin therapy failed to reduce weight in common human obesity. Consequently, interest in leptin waned for many years. However, recent studies in animal models of common obesity show that combining leptin with other weight-loss-stimulating agents, can restore hypothalamic leptin sensitivity and induce a greater reduction of body weight than after single drug treatment. Moreover, after initially reducing weight by combination therapy, leptin monotherapy proved enough to maintain reduced body weight. The strongest evidence for restoring leptin’s weight-lowering actions in common obesity to date comes from mouse studies in which leptin was combined with other pharmacotherapies. However, it is unclear at this moment whether this strategy also can be used to treat common human obesity successfully. Overcoming leptin resistance, by combining leptin with other pharmacotherapies, is a promising strategy that needs to be further investigated in humans with obesity. Full article

Prenatal alcohol exposure (PAE) leads to fetal alcohol spectrum disorder (FASD), the most common preventable cause of neurodevelopmental impairment. The classical narrative attributes the clinical picture of FASD exclusively to direct ethanol-induced brain injury. In the present review, we argue that this perspective is incomplete and leads to diagnostic errors, most often to the misdiagnosis of ADHD in children who in fact have FASD. We propose that, alongside the direct neurotoxicity of ethanol, an important and clinically under-recognized complementary mechanism is gut–brain axis dysfunction: alcohol damages the enteric nervous system and enteric glial cells, induces dysbiosis with deep deficits of butyrate and other short-chain fatty acids (SCFAs), damages the enterochromaffin cells responsible for 90% of peripheral serotonin production, and—through translocation of lipopolysaccharide (LPS) and activation of the Toll-like receptor 4 (TLR4)—sustains a neuroinflammatory brain signature. This cascade—superimposed on direct ethanol neurotoxicity—may account for the high rates of depression, anxiety, self-injurious behavior, and suicide attempts observed in individuals with FASD and for the limited efficacy of traditional interventions focused solely on the central nervous system. The 2024 Polish Institute of Mother and Child (Okulicz-Kozaryn et al.) study showed that 50.3% of pregnant women consumed alcohol, and 11% did so regularly, against only 7% who admitted so in questionnaires. The real clinical picture of children with FASD is further complicated by three factors to which we devote separate sections in this paper: prenatal co-exposure to nicotine, cannabinoids, and opioids; the loss of vertical microbiota transmission and breastfeeding in children transferred to foster care (where the prevalence of FASD is 18.8% and in children’s homes in some regions reaches up to 80%); and the substantial over-representation of preterm and small-for-gestational-age (SGA) infants (in the Hasken et al. cohort, 18.4% of children with FASD were born preterm and 51.4% were born SGA). In the final section, we present a structured, staged protocol for nutritional and microbiological intervention grounded in a hierarchy of evidence: from interventions supported by randomized controlled trials (RCT-level; choline) through interventions supported by strong mechanistic rationale and RCTs in related populations (sodium butyrate, Lactobacillus rhamnosus GG, GOS/FOS prebiotics—galacto-oligosaccharides and fructo-oligosaccharides, and omega-3 fatty acids) to experimental interventions. The protocol also covers the window before 2 years of age: we argue that, given the over-representation of preterm and SGA infants among children with FASD, the analogy to preterm infants on parenteral nutrition and to post-institutional infants applies in substantial part to the same patients, which justifies extending the indications for choline and other nutritional interventions. The paper includes a compact table of dosing proposals for each age window (from pregnancy to school-age child) and provides clinicians with concrete answers: where to start, what to avoid, and what to monitor, with explicit signposting of regulatory limitations for individual substances in Poland and the European Union. Full article

Depression is a leading cause of disability worldwide, and currently available antidepressants are limited by delayed therapeutic onset, inadequate efficacy in some patients, and adverse effects. G protein-coupled receptors (GPCRs), the largest family of membrane receptors in the central nervous system, regulate neurotransmission, neuroplasticity, neuroinflammation, stress responses, and reward processing, and are therefore important targets for antidepressant drug development. Natural products are a rich source of structurally diverse bioactive compounds, many of which show antidepressant-like effects through the modulation of GPCR-mediated signaling pathways. In this narrative review, we summarize the roles of major GPCR families implicated in depression and provide an updated overview of natural products that modulate these receptors. We particularly emphasize receptor-specific mechanisms, downstream signaling networks, and the pharmacological actions of representative natural compounds. We also highlight emerging concepts in GPCR biology, including receptor heteromerization, signaling bias, and allosteric modulation, that may create new opportunities for antidepressant discovery. Finally, we discuss current challenges related to target validation, pharmacokinetics, and clinical translation. Collectively, these insights support further investigation of natural product-derived GPCR modulators as potential leads for next-generation antidepressant development. Full article

Natural products featuring a direct σ-bond between a cyclic monoterpene and an aromatic moiety provide a vast source of biological activities, such as antimicrobial, anticancer, antiviral, anticoagulant and cannabinoid regulatory, among others. Only few methods exist for synthetically accessing such hybrid structures and their analogs, all of which are prone to limitations, most notably the reliance on sensitive organometallic intermediates and the difficulty in furnishing certain stereoisomers. An efficient, three-stage synthetic methodology is described herein, that enables the production of hybrid structures featuring a C(sp³)-C(sp²) bond between six-membered cyclic monoterpenes and aromatic moieties. This process combines: enol triflate formation from a terpenoid ketone precursor, that introduces most of the stereochemical information; Suzuki–Miyaura C-C cross-coupling of the enol triflate with a pool of (hetero)arylboronic acids, to establish the terpene–aromatic link, initially in the form of a C(sp²)-C(sp²) bond; and a stereoselective hydrogenation of the resulting adducts to afford the target compounds, establishing the stereoconfiguration of the last chiral center. Enantiomeric terpenoid scaffolds derived from menthone and trans-tetrahydrocarvone have been combined with six (6) (hetero)arylboronic acids, including medicinally relevant moieties, such as methoxyphenyl, pyridine, quinoline and benzofuran. The power of this method, apart from circumventing the need for in situ-formed sensitive organometallic intermediates, resides in providing access, for the first time, to menthyl- and trans-tetrahydrocarvoneyl-type stereoisomers, that were unattainable by any previously described method. The resulting compound library members exhibit drug-like features, based on the computational assessment of 11 selected physicochemical parameters (molecular weight, polarity, aqueous solubility, degree of unsaturation, conformational flexibility, lipophilicity, BBB permeability, skin permeability, gastrointestinal absorption, P-glycoprotein substrate behavior and Lipinski compatibility), using the platforms SwissADME, ADMETLab 3.0 and pkCSM. A computational docking study employing AutoDock Vina further identified promising candidates for targeting the known binding sites of human cannabinoid receptors CB1 and CB2, with calculated binding affinities comparable to those of established ligands. Full article

The endocannabinoid system (ECS) is increasingly recognized as a central regulator of immune homeostasis, integrating neural, metabolic, and immune signaling to maintain physiological equilibrium. This Perspective examines the “therapeutic paradox” of endocannabinoid immunomodulation, whereby anti-inflammatory and tissue-protective effects are mechanistically linked to transient immunosuppression. Although cannabinoid receptor 2 (CB2) is the primary mediator of immune regulation, growing evidence indicates that cannabinoid receptor 1 (CB1) also contributes to inflammatory control in both the central nervous system and peripheral tissues. Activation of CB2 suppresses inflammatory signaling through Gi/o-mediated inhibition of adenylate cyclase, reduced cyclic adenosine monophosphate (cAMP) signaling, and repression of nuclear factor kappa B (NF-κB)-dependent transcription. While these mechanisms limit pathological inflammation and promote tissue protection, they simultaneously attenuate innate and adaptive immune functions required for effective pathogen clearance. Across neuroinflammatory disorders, inflammatory bowel disease, hepatic injury, sepsis, cancer, and systemic inflammatory syndromes, the ECS shifts immune responses toward resolution at the cost of reduced antimicrobial readiness. We synthesize the molecular mechanisms underlying this therapeutic paradox, including macrophage polarization, lymphocyte reprogramming, and tissue-specific immune adaptations, and discuss strategies for developing endocannabinoid-based therapeutics that preserve anti-inflammatory efficacy while minimizing immunosuppressive liabilities. Full article

Cannabis sativa contains more than 120 phytocannabinoids, with Δ9-tetrahydrocannabinol (THC) and cannabidiol (CBD) being the best characterized. This review synthesizes preclinical and clinical evidence on hemp-derived extracts, cannabinoids, and active compounds. THC primarily acts as a partial agonist at cannabinoid receptor type 1 (CB1) and type 2 (CB2), producing psychoactive, appetite-stimulating, antiemetic, and analgesic effects. CBD is non-intoxicating and has a multimodal profile involving CB1 negative allosteric modulation, CB2 inverse agonism or antagonism, inhibition of anandamide inactivation, and activity at 5-HT1A receptors, transient receptor potential channels, GPR55, and peroxisome proliferator-activated receptor gamma. Preclinical models of Parkinson’s disease, Alzheimer’s disease, Huntington’s disease, epilepsy, and pain support anti-inflammatory, antioxidant, anti-excitotoxic, and glial-modulating mechanisms, but clinical translation remains uneven. The strongest evidence supports FDA-approved cannabidiol for Lennox–Gastaut syndrome, Dravet syndrome, and tuberous sclerosis complex, and THC-based agents for refractory chemotherapy-induced nausea and vomiting and AIDS-related anorexia. Moderate-certainty evidence supports nabiximols for multiple sclerosis spasticity and small benefits in selected chronic neuropathic pain populations. Evidence remains insufficient or negative for acute pain, insomnia, most psychiatric disorders, and many promoted indications. Key risks include cannabis use disorder, cognitive and psychiatric effects, cardiovascular events, sedation, high-dose CBD hepatotoxicity, and drug interactions. Rigorous, long-term, product-standardized trials are needed. Full article

Background/Objectives: Cannabinoid receptor type 2 (CB₂) agonists have attracted attention because of their potential effects on tumor-related cellular processes in different cancer models. In the present study, we investigated the effects of the selective CB₂ agonist JWH-133 on prostate cancer cell lines (LNCaP, DU-145, and PC3). Methods: Cell viability was evaluated using the MTT assay, while colony-forming capacity and migration-related responses were assessed by colony formation and wound-healing assays, respectively. In addition, the expression levels of selected cell-cycle- and apoptosis-related genes were analyzed by quantitative real-time PCR. Results: JWH-133 reduced cell viability in a time- and concentration-dependent manner, although the magnitude of this effect differed among prostate cancer cell lines. The compound also reduced colony formation in PC3 and LNCaP cells and decreased wound closure in PC3 cells under the experimental conditions used. Furthermore, JWH-133 altered the expression of several cell-cycle- and apoptosis-related genes in DU-145 and PC3 cells. Conclusions: Overall, these findings suggest that JWH-133 modulates multiple cellular responses in prostate cancer cells in a cell-line-dependent manner. However, additional mechanistic studies are required to clarify the molecular pathways underlying these effects. Full article

Endocannabinoids are lipid mediators consisting of esters, amides and ethers of long-chain polyunsaturated fatty acids. In this work, attention was focused on N-arachidonoyl tyrosine (NA-Tyr) and N-arachidonoyl dopamine (NADA), the amides of arachidonic acid with tyrosine and dopamine, respectively. NADA is an endogenous ligand of both type 1 cannabinoid receptors and type 1 vanilloid channel receptors. NADA is considered an endogenous compound with capsaicin-like activity and is distributed in several brain areas. The metabolic fate of endocannabinoids involves numerous enzymatic activities, which are only partially characterized. In particular, the biological activity of these biomolecules is terminated by enzymes with hydrolytic or oxygenase/oxidase activity. As part of this problem, we studied the oxidation of NADA and NA-Tyr mediated by mushroom tyrosinase. Our experimental data show that tyrosinase can oxidize both NADA and NA-Tyr. The oxidation of these biomolecules was also carried out in the presence of cysteine, allowing us to observe the formation of endocannabinoid/endovanilloid adducts with cysteine. These results were derived from chromatographic analyses and mass spectral experiments. During the tyrosinase-mediated oxidation in the presence of cysteine, it was possible to observe the production of a melanin-like pigment. The spectral characteristics of this pigment are consistent with those of pheomelanin, the pigment that contributes to the structure of neuromelanin. While mushroom tyrosinase serves here as a convenient biomimetic model to investigate the oxidative susceptibility of NADA and NA-Tyr, extrapolating these in vitro findings to mammalian physiology requires caution. Nevertheless, considering the neuronal distribution of these precursors and the documented, albeit debated, presence of tyrosinase-like activity in the central nervous system (CNS), these results offer a chemical rationale to further investigate whether similar oxidative pathways occur in vivo and potentially contribute to neurodegenerative mechanisms. Full article

Drug-resistant epilepsy is often associated with a neurodevelopmental disorder, focal cortical dysplasia (FCD). As demonstrated by numerous studies, defects in perisomatic inhibition may play a role in the formation of seizures. In our previous study, we observed an increase in parvalbumin-immunopositive perisomatic input. It has been observed that other perisomatic inhibitory cells expressing cannabinoid receptor type-1 (CB1R) and containing cholecystokinin (CCK) sprout in epileptic hippocampi. Therefore, we have investigated whether CB1R-immunopositive innervation has also changed in FCD. FCDIIB surgical samples were compared to controls with short post-mortem delay. The perisomatic input contacting principal cells was examined by NeuN-CB1R double immunostaining. Quantification in 3D was performed using a confocal fluorescence microscope. The results show that the CB1R-immunopositive synaptic input of principal cells is significantly larger in FCD cases. This condition may be an adaptive response to abnormal activity, or it may be a pre-existing pathology that is exacerbated by seizures. The reorganisation of the perisomatic inhibitory system has the potential to further increase the seizure probability in both cases. It should be noted that alterations in both PV- and CCK-CB1R-expressing basket cells may be implicated in seizure generation. Full article

The transcriptional regulation of the cannabinoid receptor 1 (CB1R) by promoter/enhancer elements and transcription factors is an area of cannabinoid research that has historically been understudied. To map the promoter region of the human CNR1 gene (the gene encoding CB1R), a 997-base-pair fragment from the sequence upstream of the CNR1 gene was cloned into a secreted luciferase reporter vector, and a series of deletion fragments were constructed. The transcriptional activity of these constructs was tested in human cell lines from three tissues: neuronal tissue (SHSY5Y), kidney tissue (HEK293T), and colonic epithelium (HCT116). Through this mapping, we have identified two key regulatory regions within the promoter. Increased levels of cAMP suppressed reporter expression from the full-length promoter fragment in all three cell lines, and in silico modeling predicts potential cAMP response elements (CRE) within one of the key regulatory sequences. Additionally, the minimal promoter region for CNR1 also appears to be in the second regulatory region identified, and in silico modeling predicts BRE and INR elements within this sequence. These findings begin to unravel the mechanisms by which CNR1 is transcriptionally regulated. Full article