Abstract | OBJECTIVE: METHODS: Airway epithelial cell line BEAS-2B was treated with house dust mite (HDM) to mimic allergic asthma in vitro. Lentivirus oePARK2 and siPARK2 were constructed to overexpress and knock down PARK2 expression, respectively. RT-qPCR, western blot, co-immunoprecipitation, and ubiquitination assay were performed to investigate the interaction between PARK2 and NLRP3. NLRP3 inflammasome activity, IL-1β and IL-18 secretion, pyroptosis, and epithelial barrier integrity were detected to explore the role of PARK2 in allergic asthma. RESULTS: PARK2 expression was remarkably down-regulated in HDM-treated BEAS-2B cells. In BEAS-2B cells, NLRP3 protein was reduced by PARK2 overexpression and increased by PARK2 knockdown. Interestingly, PARK2 overexpression and knockdown didn't affect NLRP3 mRNA. Co-immunoprecipitation assay showed that PARK2 interacted with NLRP3. Proteasome inhibitor MG132 abolished PARK2 overexpression-induced down-regulation of NLRP3 protein. Ubiquitination assays showed that PARK2 overexpression enhanced the ubiquitination of NLRP3. Collectively, PARK2 negatively regulates NLRP3 protein via ubiquitination. In HDM-treated BEAS-2B cells, PARK2 overexpression repressed HDM-induced NLRP3 inflammasome activation, IL-1β and IL-18 secretion, pyroptosis, and epithelial barrier dysfunction. In BEAS-2B cells, PARK2 knockdown promoted NLRP3 inflammasome activation, IL-1β and IL-18 secretion, pyroptosis, and barrier impairment, while its effects were abrogated by NLRP3 inhibitor INF39. CONCLUSION: Our study demonstrates that PARK2 attenuates HDM-induced NLRP3 inflammasome activation, the release of inflammatory cytokines, pyroptosis, and barrier impairment in airway epithelial cells by ubiquitinating NLRP3.
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Authors | Xiahui Ge, Feng Cai, Yan Shang, Feng Chi, Hua Xiao, Jing Xu, Youhui Fu, Chong Bai |
Journal | American journal of translational research
(Am J Transl Res)
Vol. 13
Issue 1
Pg. 326-335
( 2021)
ISSN: 1943-8141 [Print] United States |
PMID | 33527027
(Publication Type: Journal Article)
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Copyright | AJTR Copyright © 2021. |