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30 April 2020 : Animal Research  

Carbon Monoxide Inhibits the Expression of Proteins Associated with Intestinal Mucosal Pyroptosis in a Rat Model of Sepsis Induced by Cecal Ligation and Puncture

Hongzhou Wang1ABCDEF, Shunwen Zhang2BC, Haijun Zhao3AF, Huiyuan Qin2C, Jie Zhang3AD, Jiangtao Dong3FG, Hui Zhang1G, Xiaoling Liu1AE, Zhengyong Zhao1BF, Yanheng Zhao3BF, Meng Shao1BF, Fang Wu1AEF*, Wanjiang Zhang1AEFG

DOI: 10.12659/MSM.920668

Med Sci Monit 2020; 26:e920668

Abstract

BACKGROUND: Carbon monoxide (CO) has anti-inflammatory effects and protects the intestinal mucosal barrier in sepsis. Pyroptosis, or cell death associated with sepsis, is mediated by caspase-1 activation. This study aimed to investigate the role of CO on the expression of proteins associated with intestinal mucosal pyroptosis in a rat model of sepsis induced by cecal ligation and puncture (CLP).

MATERIAL AND METHODS: The rat model of sepsis was developed using CLP. Male Sprague-Dawley rats (n=120) were divided into six study groups: the sham group (n=20); the CLP group (n=20); the hemin group (treated with ferric chloride and heme) (n=20); the zinc protoporphyrin IX (ZnPPIX) group (n=20); the CO-releasing molecule 2 (CORM-2) group (n=20); and the inactive CORM-2 (iCORM-2) group (n=20). Hemin and CORM-2 were CO donors, and ZnPPIX was a CO inhibitor. In the six groups, the seven-day survival curves, the fluorescein isothiocyanate (FITC)-labeled dextran 4000 Da (FD-4) permeability assay, levels of intestinal pyroptosis proteins caspase-1, caspase-11, and gasdermin D (GSDMD) were measured by confocal fluorescence microscopy. Proinflammatory cytokines interleukin (IL)-18, IL-1β, and high mobility group box protein 1 (HMGB1) were measured by Western blot and enzyme-linked immunosorbent assay (ELISA).

RESULTS: CO reduced the mortality rate in rats with sepsis and reduced intestinal mucosal permeability and mucosal damage. CO also reduced the expression levels of IL-18, IL-1β, and HMGB1, and reduced pyroptosis by preventing the cleavage of caspase-1 and caspase-11.

CONCLUSIONS: In a rat model of sepsis induced by CLP, CO had a protective role by inhibiting intestinal mucosal pyroptosis.

Keywords: Carbon Monoxide, Cell Death, Intestinal Mucosa, Sepsis, Caspase 1, Cecum, Cytokines, Disease Models, Animal, Intestines, Ligation, Male, Punctures, Pyroptosis, Rats, Rats, Sprague-Dawley, Tumor Necrosis Factor-alpha

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750