PMID- 24675728 OWN - NLM STAT- MEDLINE DCOM- 20141209 LR - 20231104 IS - 1660-3397 (Electronic) IS - 1660-3397 (Linking) VI - 12 IP - 4 DP - 2014 Mar 26 TI - Vibrio vulnificus MO6-24/O lipopolysaccharide stimulates superoxide anion, thromboxane B(2), matrix metalloproteinase-9, cytokine and chemokine release by rat brain microglia in vitro. PG - 1732-56 LID - 10.3390/md12041732 [doi] AB - Although human exposure to Gram-negative Vibrio vulnificus (V. vulnificus) lipopolysaccharide (LPS) has been reported to result in septic shock, its impact on the central nervous system's innate immunity remains undetermined. The purpose of this study was to determine whether V. vulnificus MO6-24/O LPS might activate rat microglia in vitro and stimulate the release of superoxide anion (O(2)(-)), a reactive oxygen species known to cause oxidative stress and neuronal injury in vivo. Brain microglia were isolated from neonatal rats, and then treated with either V. vulnificus MO6-24/O LPS or Escherichia coli O26:B6 LPS for 17 hours in vitro. O(2)(-) was determined by cytochrome C reduction, and matrix metalloproteinase-2 (MMP-2) and MMP-9 by gelatinase zymography. Generation of cytokines tumor necrosis factor alpha (TNF-alpha), interleukin-1 alpha (IL-1alpha), IL-6, and transforming growth factor-beta 1 (TGF-beta1), chemokines macrophage inflammatory protein (MIP-1alpha)/chemokine (C-C motif) ligand 3 (CCL3), MIP-2/chemokine (C-X-C motif) ligand 2 (CXCL2), monocyte chemotactic protein-1 (MCP-1)/CCL2, and cytokine-induced neutrophil chemoattractant-2alpha/beta (CINC-2alpha/beta)/CXCL3, and brain-derived neurotrophic factor (BDNF), were determined by specific immunoassays. Priming of rat microglia by V. vulnificus MO6-24/O LPS in vitro yielded a bell-shaped dose-response curve for PMA (phorbol 12-myristate 13-acetate)-stimulated O(2)(-) generation: (1) 0.1-1 ng/mL V. vulnificus LPS enhanced O(2)(-) generation significantly but with limited inflammatory mediator generation; (2) 10-100 ng/mL V. vulnificus LPS maximized O(2)(-) generation with concomitant release of thromboxane B2 (TXB2), matrix metalloproteinase-9 (MMP-9), and several cytokines and chemokines; (3) 1000-100,000 ng/mL V. vulnificus LPS, with the exception of TXB2, yielded both attenuated O(2)(-) production, and a progressive decrease in MMP-9, cytokines and chemokines investigated. Thus concentration-dependent treatment of neonatal brain microglia with V. vulnificus MO6-24/O LPS resulted in a significant rise in O(2)(-) production, followed by a progressive decrease in O(2)(-) release, with concomitant release of lactic dehydrogenase (LDH), and generation of TXB2, MMP-9, cytokines and chemokines. We hypothesize that the inflammatory mediators investigated may be cytotoxic to microglia in vitro, by an as yet undetermined autocrine mechanism. Although V. vulnificus LPS was less potent than E. coli LPS in vitro, inflammatory mediator release by the former was clearly more efficacious. Finally, we hypothesize that should V. vulnificus LPS gain entry into the CNS, it would be possible that microglia might become activated, resulting in high levels of O(2)(-) as well as neuroinflammatory TXB2, MMP-9, cytokines and chemokines. FAU - Mayer, Alejandro M S AU - Mayer AM AD - Department of Pharmacology, Chicago College of Osteopathic Medicine, Midwestern University, 555 31st Street, Downers Grove, IL 60515, USA. amayer@midwestern.edu. FAU - Hall, Mary L AU - Hall ML AD - Department of Pharmacology, Chicago College of Osteopathic Medicine, Midwestern University, 555 31st Street, Downers Grove, IL 60515, USA. hollandmichaela@gmail.com. FAU - Holland, Michael AU - Holland M AD - Department of Pharmacology, Chicago College of Osteopathic Medicine, Midwestern University, 555 31st Street, Downers Grove, IL 60515, USA. hollandmichaela@gmail.com. FAU - De Castro, Cristina AU - De Castro C AD - Department of Chemical Sciences, University of Napoli, Via Cintia 4, 80126 Napoli, Italy. decastro@unina.it. FAU - Molinaro, Antonio AU - Molinaro A AD - Department of Chemical Sciences, University of Napoli, Via Cintia 4, 80126 Napoli, Italy. molinaro@unina.it. FAU - Aldulescu, Monica AU - Aldulescu M AD - Department of Pharmacology, Chicago College of Osteopathic Medicine, Midwestern University, 555 31st Street, Downers Grove, IL 60515, USA. monica.aldulescu@mwumail.midwestern.edu. FAU - Frenkel, Jeffrey AU - Frenkel J AD - Department of Pharmacology, Chicago College of Osteopathic Medicine, Midwestern University, 555 31st Street, Downers Grove, IL 60515, USA. jfrenk25@gmail.com. FAU - Ottenhoff, Lauren AU - Ottenhoff L AD - Department of Pharmacology, Chicago College of Osteopathic Medicine, Midwestern University, 555 31st Street, Downers Grove, IL 60515, USA. lottenhoff57@midwestern.edu. FAU - Rowley, David AU - Rowley D AD - Department of Biomedical and Pharmaceutical Sciences, College of Pharmacy, University of Rhode Island, 7 Greenhouse Road, Kingston, RI 02881, USA. drowley@mail.uri.edu. FAU - Powell, Jan AU - Powell J AD - Department of Epidemiology and Preventive Medicine, University of Maryland School of Medicine, 10 Pine St, Baltimore, MD 21201, USA. jpowell@shire.com. LA - eng PT - Comparative Study PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20140326 PL - Switzerland TA - Mar Drugs JT - Marine drugs JID - 101213729 RN - 0 (Chemokines) RN - 0 (Cytokines) RN - 0 (Lipopolysaccharides) RN - 11062-77-4 (Superoxides) RN - 54397-85-2 (Thromboxane B2) RN - EC 3.4.24.35 (Matrix Metalloproteinase 9) SB - IM MH - Animals MH - Animals, Newborn MH - Brain/immunology/metabolism/microbiology MH - Chemokines/metabolism MH - Cytokines/metabolism MH - Escherichia coli/*pathogenicity MH - Immunity, Innate MH - Lipopolysaccharides/*administration & dosage/isolation & purification MH - Matrix Metalloproteinase 9/drug effects/metabolism MH - Microglia/immunology/*metabolism/microbiology MH - Rats MH - Rats, Sprague-Dawley MH - Superoxides/metabolism MH - Thromboxane B2/metabolism MH - Vibrio vulnificus/*pathogenicity PMC - PMC4012467 EDAT- 2014/03/29 06:00 MHDA- 2014/12/15 06:00 PMCR- 2014/04/01 CRDT- 2014/03/29 06:00 PHST- 2013/12/28 00:00 [received] PHST- 2014/02/26 00:00 [revised] PHST- 2014/02/26 00:00 [accepted] PHST- 2014/03/29 06:00 [entrez] PHST- 2014/03/29 06:00 [pubmed] PHST- 2014/12/15 06:00 [medline] PHST- 2014/04/01 00:00 [pmc-release] AID - md12041732 [pii] AID - marinedrugs-12-01732 [pii] AID - 10.3390/md12041732 [doi] PST - epublish SO - Mar Drugs. 2014 Mar 26;12(4):1732-56. doi: 10.3390/md12041732.