NOX2 mediates NLRP3/ROS facilitating nasal mucosal epithelial inflammation in chronic rhinosinusitis with nasal polyps
Sijie Jiang,
Benjian Zhang,
Sihui Wen,
Shenghao Cheng,
Yingchun Shen,
Shaobing Xie,
Zhihai Xie,
Weihong Jiang
Affiliations
Sijie Jiang
Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China; Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Xiangya Hospital of Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital of Central South University, Changsha, China; Anatomy Laboratory of Division of Nose and Cranial Base, Clinical Anatomy Center of Xiangya Hospital, Central South University, Changsha, China
Benjian Zhang
Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China; Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Xiangya Hospital of Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital of Central South University, Changsha, China; Anatomy Laboratory of Division of Nose and Cranial Base, Clinical Anatomy Center of Xiangya Hospital, Central South University, Changsha, China
Sihui Wen
Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China; Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Xiangya Hospital of Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital of Central South University, Changsha, China; Anatomy Laboratory of Division of Nose and Cranial Base, Clinical Anatomy Center of Xiangya Hospital, Central South University, Changsha, China
Shenghao Cheng
Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China; Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Xiangya Hospital of Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital of Central South University, Changsha, China; Anatomy Laboratory of Division of Nose and Cranial Base, Clinical Anatomy Center of Xiangya Hospital, Central South University, Changsha, China
Yingchun Shen
John A. Burns School of Medicine, University of Hawaii, Honolulu, HI, USA
Shaobing Xie
Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China; Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Xiangya Hospital of Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital of Central South University, Changsha, China; Anatomy Laboratory of Division of Nose and Cranial Base, Clinical Anatomy Center of Xiangya Hospital, Central South University, Changsha, China; Corresponding author. Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China
Zhihai Xie
Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China; Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Xiangya Hospital of Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital of Central South University, Changsha, China; Anatomy Laboratory of Division of Nose and Cranial Base, Clinical Anatomy Center of Xiangya Hospital, Central South University, Changsha, China; Corresponding author. Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China
Weihong Jiang
Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China; Hunan Province Key Laboratory of Otolaryngology Critical Diseases, Xiangya Hospital of Central South University, Changsha, China; National Clinical Research Center for Geriatric Disorders, Xiangya Hospital of Central South University, Changsha, China; Anatomy Laboratory of Division of Nose and Cranial Base, Clinical Anatomy Center of Xiangya Hospital, Central South University, Changsha, China; Corresponding author. Department of Otolaryngology Head and Neck Surgery, Xiangya Hospital of Central South University, Changsha, China.
Background: Previous investigations have provided limited insight into the role of oxidative stress in nasal mucosa inflammation. The aim of this study was to investigate the mechanism of oxidative stress in the epithelial cells of chronic rhinosinusitis with nasal polyps CRSwNP utilizing single-cell RNA sequencing data. Methods: Single-cell RNA sequencing data from HRA000772 were used to assess oxidative stress, inflammasome activation, and nicotinamide adenine dinucleotide phosphate oxidases (NOXs) expression in epithelial cells via integrative rank-based gene set enrichment analysis. The localization of reactive oxygen species (ROS) and NOX2 in nasal mucosa and cell models was visualized using fluorescent probes and immunohistochemistry, respectively. Functional studies on NOX2 involved siRNA and plasmid transfections in vitro, while Nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3) inflammasome activity was examined using the inducer TMAO and the inhibitor MCC950. Results: Single-cell RNA sequencing data suggested an increase of oxidative stress score and NLRP3 inflammasome score in CRSwNP epithelial cells. Vitro experiments demonstrated that lipopolysaccharide could induce ROS accumulation, NLRP3 inflammasome activation and epithelial alarmin expression. MCC950 inhibited the expression of epithelia alarmin in vitro. Elevated NOX2 in CRSwNP epithelial cells was associated with increased ROS, NLRP3 inflammasome activation, and epithelial alarmin expression. NOX2-targeted siRNA inhibited these effects in vitro. Moreover, TMAO reversed the downregulation of epithelial alarmins without impacting ROS levels. Conclusion: This study highlights the crucial role of NOX2 as a key regulator of ROS accumulation and NLRP3 inflammasome activation in CRSwNP, underscoring its potential as a valuable therapeutic target for CRSwNP.
