Activation of cannabinoid-2 receptor protects against Pseudomonas aeruginosa induced acute lung injury and inflammation

• Published in: Respiratory research 2022-12, Vol.23 (1), p.326-13, Article 326
• Main Authors: Nagre, Nagaraja, Nicholson, Gregory, Cong, Xiaofei, Lockett, Janette, Pearson, Andrew C, Chan, Vincent, Kim, Woong-Ki, Vinod, K Yaragudri, Catravas, John D
• Format: Article
• Language: English
• Subjects: Acute Lung Injury - chemically induced; Acute Lung Injury - genetics; Acute Lung Injury - immunology; Acute Lung Injury - prevention & control; Acute lung injury and inflammation; Animals; Antibiotics; Bacteria; Bacterial infections; Bacterial pneumonia; Bronchus; Cannabinoid receptors; Cannabinoid-2 receptor; Cannabinoids; Cannabinoids - pharmacology; Cannabinoids - therapeutic use; Cell activation; Cell number; Chemokines; Cytokines; Diagnosis; Disease Models, Animal; Drug resistance; Health aspects; Immune system; Immunofluorescence; Immunosuppressive agents; Inflammasomes; Inflammasomes - genetics; Inflammasomes - immunology; Inflammation; Inflammation - genetics; Inflammation - immunology; Inflammation - prevention & control; Injuries; Laboratories; Lavage; Leukocytes (neutrophilic); Lungs; Mechanics; Mice; Mice, Inbred C57BL; Mortality; Neutrophils; NF-κB protein; NLR Family, Pyrin Domain-Containing 3 Protein; Nosocomial infections; Opportunist infection; Pathogens; Pneumonia; Proteins; Pseudomonas aeruginosa; Pseudomonas Infections - genetics; Pseudomonas Infections - immunology; Pseudomonas Infections - microbiology; Pseudomonas Infections - prevention & control; Receptor, Cannabinoid, CB2 - genetics; Receptor, Cannabinoid, CB2 - immunology; Receptors; Receptors, Cannabinoid; Respiratory Distress Syndrome; Risk analysis; Risk factors
• ISSN: 1465-993X; 1465-9921; 1465-993X; 1465-9921
• DOI: 10.1186/s12931-022-02253-w

Bacterial pneumonia is a major risk factor for acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). Pseudomonas aeruginosa (PA), an opportunistic pathogen with an increasing resistance acquired against multiple drugs, is one of the main causative agents of ALI and ARDS in diverse clinical settings. Given the anti-inflammatory role of the cannabinoid-2 receptor (CB2R), the effect of CB2R activation in the regulation of PA-induced ALI and inflammation was tested in a mouse model as an alternative to conventional antibiotic therapy. In order to activate CB2R, a selective synthetic agonist, JWH133, was administered intraperitoneally (i.p.) to C57BL/6J mice. Furthermore, SR144528 (a selective CB2R antagonist) was administered in combination with JWH133 to test the specificity of the CB2R-mediated effect. PA was administered intratracheally (i.t.) for induction of pneumonia in mice. At 24 h after PA exposure, lung mechanics were measured using the FlexiVent system. The total cell number, protein content, and neutrophil population in the bronchoalveolar lavage fluid (BALF) were determined. The bacterial load in the whole lung was also measured. Lung injury was evaluated by histological examination and PA-induced inflammation was assessed by measuring the levels of BALF cytokines and chemokines. Neutrophil activation (examined by immunofluorescence and immunoblot) and PA-induced inflammatory signaling (analyzed by immunoblot) were also studied. CB2R activation by JWH133 was found to significantly reduce PA-induced ALI and the bacterial burden. CB2R activation also suppressed the PA-induced increase in immune cell infiltration, neutrophil population, and inflammatory cytokines. These effects were abrogated by a CB2R antagonist, SR144528, further confirming the specificity of the CB2R-mediated effects. CB2R-knock out (CB2RKO) mice had a significantly higher level of PA-induced inflammation as compared to that in WT mice. CB2R activation diminished the excess activation of neutrophils, whereas mice lacking CB2R had elevated neutrophil activation. Pharmacological activation of CB2R significantly reduced the PA-induced NF-κB and NLRP3 inflammasome activation, whereas CB2KO mice had elevated NLRP3 inflammasome. Our findings indicate that CB2R activation ameliorates PA-induced lung injury and inflammation, thus paving the path for new therapeutic avenues against PA pneumonia.