title: Beta-naphthoflavone causes an AhR-independent inhibition of invasion and intracellular multiplication of Listeria monocytogenes in murine hepatocytes.
abstract: We recently reported a heretofore unknown role for the aryl hydrocarbon receptor in host resistance to listeriosis in mice. Hepatocytes are an important site for Listeria monocytogenes multiplication in vivo. In this study, we investigated whether activation of AhR in TIB73 murine embryonic hepatocytes affects the ingestion and intracellular multiplication of L. monocytogenes. Treatment of TIB73 cells with the AhR agonist beta-naphthoflavone (BNF) significantly inhibited the ingestion and intracellular growth of L. monocytogenes. The inhibitory effects of BNF were dose-dependent and correlated with up-regulation of CYP1A1. Surprisingly, pretreatment with AhR antagonists (3'-MNF or alpha-naphthoflavone) or knocking-down of AhR with siRNA did not abolish the inhibitory effects of BNF. Moreover, the inhibitory effects of BNF on invasion and intracellular growth of L. monocytogenes by BNF were observed in AhR-deficient (CRL-2710), or ARNT-dysfunctional (CRL-2717) Hepa cells. We also observed similar inhibitory effects of BNF treatment using primary hepatocytes recovered from AhR(+/-) or AhR(-/-) mice. Moreover, the prototypic AhR agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) did not inhibit the invasion and intracellular growth of L. monocytogenes in TIB73 cells. Mechanistic studies demonstrated that ROS, but not TNF-alpha or iNOS, plays an important role in mediating BNF-induced inhibition. In conclusion, BNF caused an AhR-independent inhibition of ingestion and intracellular multiplication of L. monocytogenes in murine hepatocytes, mediated in part by production of ROS.
authors.affiliation: Department of Pathobiological Sciences, University of Wisconsin, Madison, WI 53706, USA. lewis.shi@stjude.org
authors.forename: Lewis Zhichang
authors.initials: LZ
authors.lastname: Shi
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Czuprynski
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D051792:Basic Helix-Loop-Helix Proteins; D002460:Cell Line; D019363:Cytochrome P-450 CYP1A1 / Q000096:biosynthesis; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D055785:Gene Knockdown Techniques; D022781:Hepatocytes / Q000187:drug effects / Q000382:microbiology*; D008089:Listeria monocytogenes / Q000187:drug effects*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010587:Phagocytosis / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000276:immunology; D018336:Receptors, Aryl Hydrocarbon / Q000819:agonists* / Q000172:deficiency; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology; D019324:beta-Naphthoflavone / Q000494:pharmacology*
chemical_list: D000900:Anti-Bacterial Agents; D051792:Basic Helix-Loop-Helix Proteins; D019363:Cytochrome P-450 CYP1A1; D004791:Enzyme Inhibitors; D017382:Reactive Oxygen Species; D018336:Receptors, Aryl Hydrocarbon; D014409:Tumor Necrosis Factor-alpha; D019324:beta-Naphthoflavone; C484722:Ahr protein, mouse