NLRP3炎症小体介导单核细胞增生李斯特菌感染小鼠和巨噬细胞的细胞焦亡

Abstract
Figure/Table
References (0)
Related Citation (2)

Download: PDF (12181 KB) HTML (1 KB)
Export: BibTeX | EndNote (RIS)

Abstract Objective: To investigate the potential mechanism of Listeria monocytogenes （Lm）infectionon pyroptosis in mice and macrophages.Methods: In vivo experiments, C57B/6 mice were divided into Lm group and negative control group（NC group）, then the mice were injected with Lm 5×104 CFU/mouse and sterile PBS via tail vein, respectively. The spleen, liver and brain tissues of mice were collected 24 hours later. The pyroptosis and NLRP3 inflammasome related protein expression were detected by Western blotting. In vitro experiments were divided into Lm group, NC group, LPS+ATP group, Lm+MCC950 group and LPS+ATP+MCC950 group. Multiplicity of infection （MOI）= 10 Lm was used to infect THP-1 derived macrophages, ELISA was used to detect IL-1β from cell supernatant, quantitative real-time PCR （qRT-RCR）and Western blotting were used to detect expression of mRNA and protein about pyroptosis and NLRP3 inflammation. Immunofluorescence （IF）was used to detect ASC-speck and co-localization with NLRP3. Results: After Lm infection of mice and macrophages, the expression level of gasdermin D （GSDMD）protein was significantly increased, and gasdermin D N-terminal （GSDMD-NT）was appeared. The levels of NLRP3, caspase-1, IL-1β, and IL-18 mRNA were significantly increased after Lm infected macrophages at 1 h, 3 h, and 6 h （P<0.05）, and the increase was the most obvious at 3 h. The protein expression of NLRP3, pro-caspase-1, pro-IL-1β and cleavage fragment caspase-1 p20, IL-1β-17 in Lminfected mouse spleen, liver, brain tissue were all significantly increased （P<0.05）.In macrophages by Lm infection, the secretion of IL-1β was increased, the secretion of caspase-1 p20 and IL-1β-17 was detected in the cell supernatant, and the expression of NLRP3, pro-caspase-1, pro-IL-1β protein in the cell lysate was increased （P<0.05）, then ASC-speck was formed and co-localized with NLRP3 protein was observed by fluorescence microscope. After pretreatment with specific NLRP3 inflammasome inhibitor MCC950, GSDMD-NT of macrophages was significantly reduced （P<0.05）. Conclusion: Pyroptosis caused by Lm infection is mainly related to the activation of NLRP3 inflammasome in mice and macrophages.

Received: 25 October 2020

	Service

	E-mail this article
	Add to my bookshelf
	Add to citation manager
	E-mail Alert
	RSS
	Articles by authors

Cite this article:

URL:

http://zzs.ujs.edu.cn/xbyxb/EN/ OR http://zzs.ujs.edu.cn/xbyxb/EN/Y2021/V31/I04/307

［1］Radoshevich L, Cossart P. Listeria monocytogenes: towards a complete picture of its physiology and pathogenesis［J］. Nat Rev Microbiol, 2018, 16（1）: 32-46.

［2］Jorgensen I, Rayamajhi M, Miao EA. Programmed cell death as a defence against infection［J］. Nat Rev Immunol, 2017, 17（3）: 151-164.

［3］Swanson KV, Deng M, Ting JP. The NLRP3 inflammasome: molecular activation and regulation to therapeutics［J］. Nat Rev Immunol, 2019, 19（8）: 477-489.

［4］Wang W, Li G, Wu D, et al. Zika virus infection induces host inflammatory responses by facilitating NLRP3 inflammasome assembly and interleukin1β secretion［J］. Nat Commun, 2018, 9（1）: 106.

［5］Shi J, Zhao Y, Wang K, et al. Cleavage of GSDMD by inflammatory caspases determines pyroptotic cell death［J］. Nature, 2015, 29（7575）: 660-665.

［6］Xia X, Wang X, Zheng Y, et al. What role does pyroptosis play in microbial infection？［J］. J Cell Physiol, 2019, 234（6）: 7885-7892.

［7］Liu X, Zhang Z, Ruan J, et al. Inflammasomeactivated gasdermin D causes pyroptosis by forming membrane pores［J］. Nature, 2016, 535（7610）: 153-158.

［8］Hayward JA, Mathur A, Ngo C, et al. Cytosolic recognition of microbes and pathogens: inflammasomes in action［J］. Microbiol Mol Biol Rev, 2018, 82（4）: e00015-e00018.

［9］Lin L, Xu L, Lv W, et al. An NLRP3 inflammasometriggered cytokine storm contributes to streptococcal toxic shocklike syndrome （STSLS）［J］. PLoS Pathog, 2019, 15（6）: e1007795.

［10］Tate MD, Ong JDH, Dowling JK, et al. Reassessing the role of the NLRP3 inflammasome during pathogenic influenza A virus infection via temporal inhibition［J］. Sci Rep, 2016, 6: 27912.

［11］Cervantes J, Nagata T, Uchijima M, et al. Intracytosolic Listeria monocytogenes induces cell death through caspase1 activation in murine macrophages［J］. Cell Microbiol, 2008, 10（1）: 41-52.

［12］Wu J, FernandesAlnemri T, Alnemri ES. Involvement of the AIM2, NLRC4, and NLRP3 inflammasomes in caspase1 activation by Listeria monocytogenes［J］. J Clin Immunol, 2010, 30（5）: 693-702.

［13］Warren SE, Mao DP, Rodriguez AE, et al. Multiple nodlike receptors activate caspase 1 during Listeria monocytogenes infection［J］. J Immunol, 2008, 1（11）: 7558-7564.

［14］Kanneganti TD, zren N, BodyMalapel M, et al. Bacterial RNA and small antiviral compounds activate caspase1 through cryopyrin/Nalp3［J］. Nature, 2006, 440（7081）: 233-236.

［15］Mariathasan S, Weiss DS, Newton K, et al. Cryopyrin activates the inflammasome in response to toxins and ATP［J］. Nature, 2006, 440（7081）: 228-232.

［16］Kim S, Bauernfeind F, Ablasser A, et al. Listeria monocytogenes is sensed by the NLRP3 and AIM2 inflammasome［J］. Eur J Immunol, 2010, 40（6）: 1545-1551.

［17］Theisen E, Sauer JD.Listeria monocytogenes and the inflammasome: from cytosolic bacteriolysis to tumor immunotherapy［J］. Curr Top Microbiol Immunol, 2016, 397: 133-160.

［18］Li W, Chang Y, Liang S, et al. NLRP3 inflammasome activation contributes to Listeria monocytogenesinduced animal pregnancy failure［J］. BMC Vet Res, 2016, 12: 36.

［19］Song L, Pei L, Yao SL, et al. NLRP3 inflammasome in neurological diseases,from functions to ther