Ergolide covalently binds NLRP3 and inhibits NLRP3 inflammasome-mediated pyroptosis

  • Int Immunopharmacol. 2023 May 12;120:110292. doi: 10.1016/j.intimp.2023.110292.
Miao Ren  1 ,  Jiahao Chen  2 ,  Haowen Xu  2 ,  Weifeng Li  2 ,  Tingting Wang  3 ,  Zhanghuan Chi  4 ,  Yi Lin  4 ,  Anqi Zhang  5 ,  Gaozhi Chen  2 ,  Xu Wang  2 ,  Xiaoyu Sun  2 ,  Guang Liang  6 ,  Junlu Wang  7 ,  Wu Luo  8
Affiliations
  • 1. The Department of Anesthesiology and Operation Room, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
  • 2. Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
  • 3. Taizhou Hospital of Zhejiang Province Affiliated to Wenzhou Medical University, Taizhou, Zhejiang 317099, China.
  • 4. Wenzhou Third Clinical College of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
  • 5. The Department of Anesthesiology and Operation Room, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
  • 6. Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China; School of Pharmaceutical Sciences, Hangzhou Medical College, Hangzhou, Zhejiang 311399, China. Electronic address: [email protected].
  • 7. The Department of Anesthesiology and Operation Room, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China. Electronic address: [email protected].
  • 8. Medical Research Center, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China. Electronic address: [email protected].
PMID: 37182452 DOI: 10.1016/j.intimp.2023.110292
Abstract

Background: NLR family pyrin domain-containing 3 (NLRP3)-mediated Pyroptosis plays a key role in various acute and chronic inflammatory diseases. Targeted inhibition of NLRP3-mediated Pyroptosis may be a potential therapeutic strategy for various inflammatory diseases. Ergolide (ERG) is a sesquiterpene lactone natural product derived from the traditional Chinese medicinal herb, Inula britannica. ERG has been shown to have anti-inflammatory and anti-cancer activities, but the target is remains unknown.

Hypothesis/purpose: This study performed an in-depth investigation of the anti-inflammatory mechanism of ERG in NLRP3-mediated Pyroptosis and NLPR3 inflammasome related sepsis and acute lung injury model.

Methods: ELISA and Western blot were used to determine the IL-1β and P20 levels. Co-immunoprecipitation assays were used to detect the interaction between proteins. Drug affinity response target stability (DARTS) assays were used to explore the potential target of ERG. C57BL/6J mice were intraperitoneally injected with E. coli DH5α (2 × 109 CFU/mouse) to establish a sepsis model. Acute lung injury was induced by intratracheal administrationof lipopolysaccharide in wild-type mice and NLRP3 knockout mice with or without ERG treatment.

Results: We showed that ERG is an efficient inhibitor of NLRP3-mediated Pyroptosis in the first and second signals of NLRP3 inflammasome activation. Furthermore, we demonstrated that ERG irreversibly bound to the NACHT domain of NLRP3 to prevent the assembly and activation of the NLRP3 inflammasome. ERG remarkably improved the survival rate of wild-type septic mice. In lipopolysaccharide-induced acute lung injury model, ERG alleviated acute lung injury of wild-type mice but not NLRP3 knockout mice.

Conclusion: Our results revealed that the anti-pyroptosis effect of ERG are dependent on NLRP3 and NLRP3 NACHT domain is ERG's direct target. Therefore, ERG can serve as a precursor drug for the development of novel NLRP3 inhibitors to treat NLRP3 inflammasome mediated inflammatory diseases.

Keywords
Acute lung injury; Ergolide; NACHT domain; NLRP3 inflammsome; Pyroptosis; Sepsis.
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