Carbon monoxide releasing molecule-3 improves myocardial function in mice with sepsis by inhibiting NLRP3 inflammasome activation in cardiac fibroblasts.
The NLRP3 inflammasome is an intracellular multiple-protein complex that controls the maturation and release of interleukin (IL)-1β and IL-18. Endogenous carbon monoxide (CO) is anti-inflammatory. The aim of this study was to assess the effects/mechanisms of CO-releasing molecule-3 (CORM-3)-dependent modulation of the NLRP3 inflammasome in cardiac fibroblasts (CF) and its effect on myocardial function in sepsis. CF were treated with CORM-3 or inactive CORM-3 (iCORM-3) before NLRP3 inflammasome priming with lipopolysaccharides (LPS) or following activation with adenosine triphosphate (ATP). In parallel, cardiomyocytes (CM) were challenged with supernatants of LPS/ATP-stimulated CF or a cytokine mixture (Cyto-mix) containing IL-1β, IL-18, and HMGB1. In vivo, mice were treated with CORM-3 before or after LPS to induce sepsis (endotoxemia). Pretreatment of CF with CORM-3 prevented an LPS-induced increase in NLRP3 and pro-IL-1β expression. Treatment of CF with CORM-3 before ATP prevented ATP-induced activation of the NLRP3 inflammasome. Challenging CF with LPS/ATP promoted NLRP3 interactions with adaptor ASC (apoptosis-associated speck-like protein containing a caspase-recruitment domain), which was prevented by CORM-3. Challenging CM with supernatants of CF with LPS/ATP or Cyto-mix (IL-1β, IL-18, and HMGB1) resulted in CM apoptosis, which was attenuated with either a CORM-3 or IL-1 receptor antagonist. Finally, myocardial NLRP3 inflammasome activation and myocardial dysfunction in septic mice were abolished by CORM-3. In NLRP3-deficient mice with sepsis, CORM-3 did not show additional benefits in improving myocardial function. Our results indicate that CORM-3 suppresses NLRP3 inflammasome activation by blocking NLRP3 interactions with the adaptor protein ASC and attenuates myocardial dysfunction in mice with sepsis.
Division of Cardiology, Department of Medicine, The Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu, People's Republic of China.|Critical Illness Research, Lawson Health Research Institute, 800 Commissioners Road E., VRL Rm A6-138, London, ON, N6A 4G5, Canada.
Wenbo
W
Zhang
Division of Cardiology, Department of Medicine, The Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu, People's Republic of China.|Critical Illness Research, Lawson Health Research Institute, 800 Commissioners Road E., VRL Rm A6-138, London, ON, N6A 4G5, Canada.
Aibin
A
Tao
Critical Illness Research, Lawson Health Research Institute, 800 Commissioners Road E., VRL Rm A6-138, London, ON, N6A 4G5, Canada.
Ting
T
Lan
Critical Illness Research, Lawson Health Research Institute, 800 Commissioners Road E., VRL Rm A6-138, London, ON, N6A 4G5, Canada.
Gediminas
G
Cepinskas
Critical Illness Research, Lawson Health Research Institute, 800 Commissioners Road E., VRL Rm A6-138, London, ON, N6A 4G5, Canada.|Critical Care Western, Schulich School of Medicine and Dentistry, Western University, London, ON, Canada.
Raymond
R
Kao
Critical Illness Research, Lawson Health Research Institute, 800 Commissioners Road E., VRL Rm A6-138, London, ON, N6A 4G5, Canada.|Critical Care Western, Schulich School of Medicine and Dentistry, Western University, London, ON, Canada.
Claudio M
CM
Martin
Division of Cardiology, Department of Medicine, The Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu, People's Republic of China. trui@uwo.ca.|Critical Illness Research, Lawson Health Research Institute, 800 Commissioners Road E., VRL Rm A6-138, London, ON, N6A 4G5, Canada. trui@uwo.ca.|Critical Care Western, Schulich School of Medicine and Dentistry, Western University, London, ON, Canada. trui@uwo.ca.|Department of Medicine, Schulich School of Medicine and Dentistry, Western University, London, ON, Canada. trui@uwo.ca.|Department of Pathology and Laboratory Medicine, Schulich School of Medicine and Dentistry, Western University, London, ON, Canada. trui@uwo.ca.
Tao
T
Rui
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D015153:Blotting, Western; D002248:Carbon Monoxide / Q000494:pharmacology; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006321:Heart / Q000187:drug effects; D047468:Immunoprecipitation; D058847:Inflammasomes / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009206:Myocardium / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000378:metabolism*; D009942:Organometallic Compounds / Q000378:metabolism / Q000494:pharmacology*; D018805:Sepsis / Q000150:complications* / Q000378:metabolism
D058847:Inflammasomes; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C492716:Nlrp3 protein, mouse; D009942:Organometallic Compounds; C476530:tricarbonylchloro(glycinato)ruthenium(II); D002248:Carbon Monoxide
Carbon monoxide; Cardiac fibroblast–cardiomyocyte interaction; Myocardial dysfunction; NLRP3 inflammasome; Sepsis