title: NLRP3 inflammasome as a target of berberine in experimental murine liver injury: interference with P2X7 signalling.
abstract: Berberine (BRB) is commonly used in herbal medicine, but its mechanisms of action are poorly understood. In the present study, we tested BRB in steatohepatitis induced by a methionine- and choline-deficient (MCD) diet, in acute acetaminophen intoxication and in cultured murine macrophages. BRB markedly improved parameters of liver injury and necroinflammation induced by the MCD diet, although increased mortality was observed by mechanisms independent of bacterial infections or plasma levels of BRB. The MCD diet induced up-regulation of all components of the NLRP3 (NACHT, LRR and PYD domain-containing protein 3) inflammasome, and increased hepatic levels of mature IL-1β (interleukin 1β). All of these parameters were significantly reduced in mice treated with BRB. In mice administered an acetaminophen overdose, a model dependent on inflammasome activation, BRB reduced mortality and ALT (alanine aminotransferase) elevation, and limited the expression of inflammasome components. In vitro, LPS (lipopolysaccharide)-induced activation of NLRP3 inflammasome in RAW264.7 murine macrophages was markedly decreased by pre-incubation with BRB. BRB significantly limited the activation of the purinergic receptor P2X7, involved in the late phases of inflammasome activation. Upon P2X7 knockdown, the ability of BRB to block LPS-induced secretion of IL-1β was lost. These data indicate that administration of BRB ameliorates inflammation and injury in two unrelated murine models of liver damage. We demonstrate for the first time that BRB interferes with activation of the NLRP3 inflammasome pathway in vivo and in vitro, through a mechanism based on interference with activation of P2X7, a purinergic receptor involved in inflammasome activation.
authors.affiliation: Dipartimento di Medicina Sperimentale e Clinica, University of Florence, Florence, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Vivoli
authors.affiliation: Dipartimento di Medicina Sperimentale e Clinica, University of Florence, Florence, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Cappon
authors.affiliation: Dipartimento di Scienze Biomediche, Sperimentali e Cliniche "Mario Serio", University of Florence, Florence, Italy Centro di Ricerca Denothe, University of Florence, Florence, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Milani
authors.affiliation: Dipartimento di Medicina Sperimentale e Clinica, University of Florence, Florence, Italy.
authors.forename: Benedetta
authors.initials: B
authors.lastname: Piombanti
authors.affiliation: Dipartimento di Medicina Sperimentale e Clinica, University of Florence, Florence, Italy.
authors.forename: Angela
authors.initials: A
authors.lastname: Provenzano
authors.affiliation: Dipartimento di Scienze Cliniche e Biologiche, University of Turin, Turin, Italy.
authors.forename: Erica
authors.initials: E
authors.lastname: Novo
authors.affiliation: Dipartimento di Neuroscienze, Psicologia, Area del Farmaco e Salute del Bambino, University of Florence, Florence, Italy.
authors.forename: Alessio
authors.initials: A
authors.lastname: Masi
authors.affiliation: Dipartimento di Medicina Sperimentale e Clinica, University of Florence, Florence, Italy.
authors.forename: Nadia
authors.initials: N
authors.lastname: Navari
authors.affiliation: Dipartimento di Neuroscienze, Psicologia, Area del Farmaco e Salute del Bambino, University of Florence, Florence, Italy.
authors.forename: Roberto
authors.initials: R
authors.lastname: Narducci
authors.affiliation: Dipartimento di Neuroscienze, Psicologia, Area del Farmaco e Salute del Bambino, University of Florence, Florence, Italy.
authors.forename: Guido
authors.initials: G
authors.lastname: Mannaioni
authors.affiliation: Dipartimento di Scienze della Salute, University of Florence, Florence, Italy.
authors.forename: Gloriano
authors.initials: G
authors.lastname: Moneti
authors.affiliation: Department of Gastroenterology (LIM07), University of São Paulo School of Medicine, São Paulo, Brazil.
authors.forename: Claudia P
authors.initials: CP
authors.lastname: Oliveira
authors.affiliation: Dipartimento di Scienze Cliniche e Biologiche, University of Turin, Turin, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Parola
authors.affiliation: Dipartimento di Medicina Sperimentale e Clinica, University of Florence, Florence, Italy Centro di Ricerca Denothe, University of Florence, Florence, Italy fabio.marra@unifi.it.
authors.forename: Fabio
authors.initials: F
authors.lastname: Marra
mesh_terms: D000082:Acetaminophen / Q000009:adverse effects*; D000818:Animals; D001599:Berberine / Q000008:administration & dosage*; D056486:Chemical and Drug Induced Liver Injury / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D058847:Inflammasomes / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D053583:Interleukin-1beta / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D058847:Inflammasomes; D053583:Interleukin-1beta; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C492716:Nlrp3 protein, mouse; D001599:Berberine; D000082:Acetaminophen
keywords: LPS; acetaminophen toxicity; chemokines; cytokines; inflammation; macrophages; non-alcoholic steatohepatitis