Advertisement

Two non-psychoactive cannabinoids reduce intracellular lipid levels and inhibit hepatosteatosis

  • Cristoforo Silvestri
    Affiliations
    Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Via Campi Flegrei 34, 80078, Pozzuoli, Naples, Italy
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally to this work.
    Debora Paris
    Footnotes
    † These authors contributed equally to this work.
    Affiliations
    Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Via Campi Flegrei 34, 80078, Pozzuoli, Naples, Italy
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally to this work.
    Andrea Martella
    Footnotes
    † These authors contributed equally to this work.
    Affiliations
    Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Via Campi Flegrei 34, 80078, Pozzuoli, Naples, Italy
    Search for articles by this author
  • Dominique Melck
    Affiliations
    Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Via Campi Flegrei 34, 80078, Pozzuoli, Naples, Italy
    Search for articles by this author
  • Irene Guadagnino
    Affiliations
    Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Via Campi Flegrei 34, 80078, Pozzuoli, Naples, Italy
    Search for articles by this author
  • Mike Cawthorne
    Affiliations
    Institute of Translational Medicine, Clore Laboratory, University of Buckingham, Buckingham, UK
    Search for articles by this author
  • Andrea Motta
    Affiliations
    Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Via Campi Flegrei 34, 80078, Pozzuoli, Naples, Italy
    Search for articles by this author
  • Vincenzo Di Marzo
    Correspondence
    Corresponding author. Address: Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Via Campi Flegrei 34, Comprensorio Olivetti, 80078, Pozzuoli (NA), Italy. Tel.: +39 081 8675093; fax: +39 081 8041770.
    Affiliations
    Institute of Biomolecular Chemistry, Consiglio Nazionale delle Ricerche, Via Campi Flegrei 34, 80078, Pozzuoli, Naples, Italy
    Search for articles by this author
  • Author Footnotes
    † These authors contributed equally to this work.
Published:January 13, 2015DOI:https://doi.org/10.1016/j.jhep.2015.01.001

      Background & Aims

      Obesity and associated metabolic syndrome have quickly become a pandemic and a major detriment to global human health. The presence of non-alcoholic fatty liver disease (NAFLD; hepatosteatosis) in obesity has been linked to the worsening of the metabolic syndrome, including the development of insulin resistance and cardiovascular disease. Currently, there are few options to treat NAFLD, including life style changes and insulin sensitizers. Recent evidence suggests that the cannabinoids Δ9-tetrahydrocannabivarin (THCV) and cannabidiol (CBD) improve insulin sensitivity; we aimed at studying their effects on lipid levels.

      Methods

      The effects of THCV and CBD on lipid levels were examined in a variety of in vitro and in vivo systems, with special emphasis on models of hepatosteatosis. Transcriptional, post-translational and metabolomic changes were assayed.

      Results

      THCV and CBD directly reduce accumulated lipid levels in vitro in a hepatosteatosis model and adipocytes. Nuclear magnetic resonance- (NMR) based metabolomics confirmed these results and further identified specific metabolic changes in THCV and CBD-treated hepatocytes. Treatment also induced post-translational changes in a variety of proteins such as CREB, PRAS40, AMPKa2 and several STATs indicating increased lipid metabolism and, possibly, mitochondrial activity. These results are supported by in vivo data from zebrafish and obese mice indicating that these cannabinoids are able to increase yolk lipid mobilization and inhibit the development of hepatosteatosis respectively.

      Conclusions

      Our results suggest that THCV and CBD might be used as new therapeutic agents for the pharmacological treatment of obesity- and metabolic syndrome-related NAFLD/hepatosteatosis.

      Graphical abstract

      Abbreviations:

      NAFLD (non-alcoholic fatty liver disease), NMR (nuclear magnetic resonance), THCV (Δ9-tetrahydrocannabivarin), CBD (cannabidiol), AEA (anandamide), 2-AG (2-arachidonoylglycerol), CB1/2 (Cannabinoid receptor 1/2), THC (Δ9-tetra-hydrocannabinol), HHL-5 (human hepatocyte line-5), OA (oleic acid), ECS (endocannabinoid system), TG (triglyceride), TRPV1 (transient receptor potential vanilloid type-1), Cho (Choline), PC (phosphocholine), FA (fatty acid), GSH (glutathione), ATP (adenine triphosphate), NAD (nicotinamide adenine dinucleotide), FChol (free cholesterol), GK (Glycerol kinase), AMPK (5′-AMP-activated protein kinase), OPLS-DA (Orthogonal Projection to Latent Structure Discriminant Analysis)

      Keywords

      To read this article in full you will need to make a payment
      Subscribe to Journal of Hepatology
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

        • Ng M.
        • Fleming T.
        • Robinson M.
        • Thomson B.
        • Graetz N.
        • Margono C.
        • et al.
        Global, regional, and national prevalence of overweight and obesity in children and adults during 1980–2013: a systematic analysis for the Global Burden of Disease Study 2013.
        Lancet. 2014; 384: 766-768
        • Postic C.
        • Girard J.
        Contribution of de novo fatty acid synthesis to hepatic steatosis and insulin resistance: lessons from genetically engineered mice.
        J Clin Invest. 2008; 118: 829-838
        • Vanni E.
        • Bugianesi E.
        • Kotronen A.
        • De Minicis S.
        • Yki-Jarvinen H.
        • Svegliati-Baroni G.
        From the metabolic syndrome to NAFLD or vice versa?.
        Dig Liver Dis. 2010; 42: 320-330
        • Silvestri C.
        • Di Marzo V.
        The endocannabinoid system in energy homeostasis and the etiopathology of metabolic disorders.
        Cell Metab. 2013; 17: 475-490
        • Jourdan T.
        • Djaouti L.
        • Demizieux L.
        • Gresti J.
        • Verges B.
        • Degrace P.
        CB1 antagonism exerts specific molecular effects on visceral and subcutaneous fat and reverses liver steatosis in diet-induced obese mice.
        Diabetes. 2010; 59: 926-934
        • Jeong W.I.
        • Osei-Hyiaman D.
        • Park O.
        • Liu J.
        • Batkai S.
        • Mukhopadhyay P.
        • et al.
        Paracrine activation of hepatic CB1 receptors by stellate cell-derived endocannabinoids mediates alcoholic fatty liver.
        Cell Metab. 2008; 7: 227-235
        • Liu J.
        • Zhou L.
        • Xiong K.
        • Godlewski G.
        • Mukhopadhyay B.
        • Tam J.
        • et al.
        Hepatic cannabinoid receptor-1 mediates diet-induced insulin resistance via inhibition of insulin signaling and clearance in mice.
        Gastroenterology. 2012; 142: e1211
        • McPartland J.M.
        • Duncan M.
        • Di Marzo V.
        • Pertwee R.
        Are cannabidiol and Delta- tetrahydrocannabivarin negative modulators of the endocannabinoid system? A systematic review.
        Br J Pharmacol. 2014; 172: 737-753
        • De Petrocellis L.
        • Ligresti A.
        • Moriello A.S.
        • Allara M.
        • Bisogno T.
        • Petrosino S.
        • et al.
        Effects of cannabinoids and cannabinoid-enriched Cannabis extracts on TRP channels and endocannabinoid metabolic enzymes.
        Br J Pharmacol. 2011; 163: 1479-1494
        • Palmer R.K.
        • Lunn C.A.
        TRP channels as targets for therapeutic intervention in obesity: focus on TRPV1 and TRPM5.
        Curr Top Med Chem. 2013; 13: 247-257
        • Ignatowska-Jankowska B.
        • Jankowski M.M.
        • Swiergiel A.H.
        Cannabidiol decreases body weight gain in rats: involvement of CB2 receptors.
        Neurosci Lett. 2011; 490: 82-84
        • Yang L.
        • Rozenfeld R.
        • Wu D.
        • Devi L.A.
        • Zhang Z.
        • Cederbaum A.
        Cannabidiol protects liver from binge alcohol-induced steatosis by mechanisms including inhibition of oxidative stress and increase in autophagy.
        Free Radic Biol Med. 2014; 68: 260-267
        • Wargent E.T.
        • Zaibi M.S.
        • Silvestri C.
        • Hislop D.C.
        • Stocker C.J.
        • Stott C.G.
        • et al.
        The cannabinoid Delta(9)-tetrahydrocannabivarin (THCV) ameliorates insulin sensitivity in two mouse models of obesity.
        Nutr Diab. 2013; 3: e68
      1. Results from GW Phase IIa Study Identify Promising New Oral Cannabinoid Anti-Diabetic Treatment. [cited; Available from: <http://www.gwpharm.com/Results%20from%20GW%20Phase%20IIa%20Study%20Identify%20Promising%20New%20Oral%20Cannabinoid%20Anti-Diabetic%20Treatment.aspx>.

        • Lindon J.C.
        • Nicholson J.K.
        • Holmes E.
        • Keun H.C.
        • Craig A.
        • Pearce J.T.
        • et al.
        Summary recommendations for standardization and reporting of metabolic analyses.
        Nat Biotechnol. 2005; 23: 833-838
      2. Eriksson L, Johansson E, AB. U, Academy U, Kettaneh-Wold N, Wold S. Multi- and megavariate data analysis: principles and applications: Umetrics Academy; 2001.

        • Silvestri C.
        • Di Marzo V.
        Second generation CB1 receptor blockers and other inhibitors of peripheral endocannabinoid overactivity and the rationale of their use against metabolic disorders.
        Expert Opin Investig Drugs. 2012; 21: 1309-1322
      3. Cascio MG, Marini, P, Bolognini, D, Pertwee, RG The 11-hydroxy Metabolite Of Δ9-Tetrahydrocannabivarin Behaves As An Apparent CB1 and CB2 Receptor Neutral Antagonist. In: 6th European Workshop on Cannabinoid Research. Dublin: British Pharmacological Society; 2013.

        • Morino K.
        • Petersen K.F.
        • Shulman G.I.
        Molecular mechanisms of insulin resistance in humans and their potential links with mitochondrial dysfunction.
        Diabetes. 2006; 55: S9-S15
        • Herzig S.
        • Hedrick S.
        • Morantte I.
        • Koo S.H.
        • Galimi F.
        • Montminy M.
        CREB controls hepatic lipid metabolism through nuclear hormone receptor PPAR-gamma.
        Nature. 2003; 426: 190-193
        • Erion D.M.
        • Ignatova I.D.
        • Yonemitsu S.
        • Nagai Y.
        • Chatterjee P.
        • Weismann D.
        • et al.
        Prevention of hepatic steatosis and hepatic insulin resistance by knockdown of cAMP response element-binding protein.
        Cell Metab. 2009; 10: 499-506
        • Viollet B.
        • Foretz M.
        • Guigas B.
        • Horman S.
        • Dentin R.
        • Bertrand L.
        • et al.
        Activation of AMP-activated protein kinase in the liver: a new strategy for the management of metabolic hepatic disorders.
        J Physiol. 2006; 574: 41-53
        • Liao C.C.
        • Ou T.T.
        • Huang H.P.
        • Wang C.J.
        The inhibition of oleic acid induced hepatic lipogenesis and the promotion of lipolysis by caffeic acid via up-regulation of AMP-activated kinase.
        J Sci Food Agric. 2014; 94: 1154-1162
        • Jelenik T.
        • Rossmeisl M.
        • Kuda O.
        • Jilkova Z.M.
        • Medrikova D.
        • Kus V.
        • et al.
        AMP-activated protein kinase alpha2 subunit is required for the preservation of hepatic insulin sensitivity by n-3 polyunsaturated fatty acids.
        Diabetes. 2010; 59: 2737-2746
        • Shahidi M.
        • Tay E.S.
        • Read S.A.
        • Ramezani-Moghadam M.
        • Chayama K.
        • George J.
        • et al.
        Endocannabinoid CB1 antagonists inhibit hepatitis C virus production, providing a novel class of antiviral host-targeting agents.
        J Gen Virol. 2014; 95: 2468-2479
        • Reiss C.S.
        Cannabinoids and viral infections.
        Pharmaceuticals (Basel). 2010; 3: 1873-1886
        • Miller A.M.
        • Wang H.
        • Bertola A.
        • Park O.
        • Horiguchi N.
        • Ki S.H.
        • et al.
        Inflammation-associated interleukin-6/signal transducer and activator of transcription 3 activation ameliorates alcoholic and nonalcoholic fatty liver diseases in interleukin-10-deficient mice.
        Hepatology. 2011; 54: 846-856
        • Barclay J.L.
        • Nelson C.N.
        • Ishikawa M.
        • Murray L.A.
        • Kerr L.M.
        • McPhee T.R.
        • et al.
        GH-dependent STAT5 signaling plays an important role in hepatic lipid metabolism.
        Endocrinology. 2011; 152: 181-192
        • Iff J.
        • Wang W.
        • Sajic T.
        • Oudry N.
        • Gueneau E.
        • Hopfgartner G.
        • et al.
        Differential proteomic analysis of STAT6 knockout mice reveals new regulatory function in liver lipid homeostasis.
        J Proteome Res. 2009; 8: 4511-4524
        • Yang L.
        • Zhang Y.
        • Wang L.
        • Fan F.
        • Zhu L.
        • Li Z.
        • et al.
        Amelioration of high fat diet induced liver lipogenesis and hepatic steatosis by interleukin-22.
        J Hepatol. 2010; 53: 339-347
        • Wegrzyn J.
        • Potla R.
        • Chwae Y.J.
        • Sepuri N.B.
        • Zhang Q.
        • Koeck T.
        • et al.
        Function of mitochondrial Stat3 in cellular respiration.
        Science. 2009; 323: 793-797
        • Bravard A.
        • Vial G.
        • Chauvin M.A.
        • Rouille Y.
        • Bailleul B.
        • Vidal H.
        • et al.
        FTO contributes to hepatic metabolism regulation through regulation of leptin action and STAT3 signalling in liver.
        Cell Commun Signal. 2014; 12: 4
        • Jiao P.
        • Feng B.
        • Li Y.
        • He Q.
        • Xu H.
        Hepatic ERK activity plays a role in energy metabolism.
        Mol Cell Endocrinol. 2013; 375: 157-166
        • Wang Z.
        • Yao T.
        • Song Z.
        Involvement and mechanism of DGAT2 upregulation in the pathogenesis of alcoholic fatty liver disease.
        J Lipid Res. 2010; 51: 3158-3165
        • Park E.J.
        • Lee J.H.
        • Yu G.Y.
        • He G.
        • Ali S.R.
        • Holzer R.G.
        • et al.
        Dietary and genetic obesity promote liver inflammation and tumorigenesis by enhancing IL-6 and TNF expression.
        Cell. 2010; 140: 197-208
        • Mukhopadhyay P.
        • Rajesh M.
        • Horvath B.
        • Batkai S.
        • Park O.
        • Tanchian G.
        • et al.
        Cannabidiol protects against hepatic ischemia/reperfusion injury by attenuating inflammatory signaling and response, oxidative/nitrative stress, and cell death.
        Free Radic Biol Med. 2011; 50: 1368-1381