title: Modulation of thrombin-induced neuroinflammation in BV-2 microglia by carbon monoxide-releasing molecule 3.
abstract: Carbon monoxide-releasing molecules are emerging as a new class of pharmacological agents that regulate important cellular function by liberating CO in biological systems. Here, we examined the role of carbon monoxide-releasing molecule 3 (CORM-3) in modulating neuroinflammatory responses in BV-2 microglial cells, considering its practical application as a novel therapeutic alternative in the treatment of stroke. BV-2 microglia cells were incubated for 24 h in normoxic conditions with thrombin alone or in combination with interferon-gamma to simulate the inflammatory response. Cells were also subjected to 12 h of hypoxia and reoxygenated for 24 h in the presence of thrombin and interferon-gamma. In both set of experiments, the anti-inflammatory action of CORM-3 was evaluated by assessing its effect on nitric oxide production (nitrite levels) and tumor necrosis factor (TNF)-alpha release. CORM-3 (75 microM) did not show any cytotoxicity and markedly attenuated the inflammatory response to thrombin and interferon-gamma in normoxia and to a lesser extent in hypoxia as evidenced by a reduction in nitrite levels and TNF-alpha production. Inactive CORM-3, which does not liberate CO and is used as a negative control, failed to prevent the increase in inflammatory mediators. Blockade of endogenous CO production by tin protoporphyrin-IX did not change the anti-inflammatory activity of CORM-3, suggesting that CO liberated from the compound is responsible for the observed effects. In addition, inhibition of the mitogen-activated protein kinases phosphatidyl inositol 3 kinase and extracellular signal-regulated kinase amplified the anti-inflammatory effect of CORM-3. These results suggest that the anti-inflammatory activity of CORM-3 could be exploited to mitigate microglia activity in stroke and other neuroinflammatory diseases.
authors.affiliation: Vascular Biology Unit, Department of Surgical Research, Northwick Park Institute for Medical Research, Harrow, Middlesex HA1 3UJ, UK.
authors.forename: Mohamed G
authors.initials: MG
authors.lastname: Bani-Hani
authors.forename: David
authors.initials: D
authors.lastname: Greenstein
authors.forename: Brian E
authors.initials: BE
authors.lastname: Mann
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Green
authors.forename: Roberto
authors.initials: R
authors.lastname: Motterlini
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002248:Carbon Monoxide / Q000627:therapeutic use; D015687:Cell Hypoxia; D002478:Cells, Cultured; D006419:Heme Oxygenase (Decyclizing) / Q000502:physiology; D007371:Interferon-gamma / Q000494:pharmacology; D020935:MAP Kinase Signaling System; D051379:Mice; D017628:Microglia / Q000187:drug effects* / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D009573:Nitrites / Q000378:metabolism; D009942:Organometallic Compounds / Q000494:pharmacology* / Q000627:therapeutic use; D020521:Stroke / Q000188:drug therapy; D013917:Thrombin / Q000633:toxicity*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D000893:Anti-Inflammatory Agents; D009573:Nitrites; D009942:Organometallic Compounds; D014409:Tumor Necrosis Factor-alpha; C476530:tricarbonylchloro(glycinato)ruthenium(II); D009569:Nitric Oxide; D002248:Carbon Monoxide; D007371:Interferon-gamma; D006419:Heme Oxygenase (Decyclizing); D013917:Thrombin