Ethanol extract of Chrysanthemum zawadskii inhibits the NLRP3 inflammasome by suppressing ASC oligomerization in macrophages

Ah-Ra Jang; Ha-Nul Lee; Jung Joo Hong; Young-Min Kim; Jong-Hwan Park

doi:10.3892/etm.2023.11827

Ethanol extract of Chrysanthemum zawadskii inhibits the NLRP3 inflammasome by suppressing ASC oligomerization in macrophages

Exp Ther Med. 2023 Feb 7;25(3):128. doi: 10.3892/etm.2023.11827. eCollection 2023 Mar.

Authors

Ah-Ra Jang^{1

2}, Ha-Nul Lee³, Jung Joo Hong⁴, Young-Min Kim³, Jong-Hwan Park^{1

2}

Affiliations

¹ Laboratory of Animal Medicine, College of Veterinary Medicine and Animal Medical Institute, Chonnam National University, Gwangju 61186, Republic of Korea.
² NodCure, Inc., Gwangju 61186, Republic of Korea.
³ Department of Food Science & Technology and Brain Korea 21 Plus Program, Chonnam National University, Gwangju 61186, Republic of Korea.
⁴ National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, Chungcheongbuk 28116, Republic of Korea.

Abstract

Chrysanthemum zawadskii (C. zawadskii) is used in traditional East Asian medicine for the treatment of various diseases, including inflammatory disease. However, it has remained unclear whether extracts of C. zawadskii inhibit inflammasome activation in macrophages. The present study assessed the inhibitory effect of an ethanol extract of C. zawadskii (CZE) on the activation of the inflammasome in macrophages and the underlying mechanism. Bone marrow-derived macrophages were obtained from wild-type C57BL/6 mice. The release of IL-1β and lactate dehydrogenase in response to nucleotide-binding oligomerization domain-like receptor (NLR) family pyrin domain containing 3 (NLRP3) inflammasome activators, such as ATP, nigericin and monosodium urate (MSU) crystals, was significantly decreased by CZE in lipopolysaccharide (LPS)-primed BMDMs. Western blotting revealed that CZE inhibited ATP-induced caspase-1 cleavage and IL-1β maturation. To investigate whether CZE inhibits the priming step of the NLRP3 inflammasome, we confirmed the role of CZE at the gene level using RT-qPCR. CZE also downregulated the gene expression of NLRP3 and pro-IL-1β as well as NF-κB activation in BMDMs in response to LPS. Apoptosis-associated speck-like protein containing a caspase-recruitment domain (CARD) oligomerization and speck formation by NLRP3 inflammasome activators were suppressed by CZE. By contrast, CZE did not affect NLR family CARD domain-containing protein 4 or absent in melanoma 2 inflammasome activation in response to Salmonella typhimurium and poly(dA:dT) in LPS-primed BMDMs, respectively. The results revealed that three key components of CZE, namely linarin, 3,5-dicaffeoylquinic acid and chlorogenic acid, decreased IL-1β secretion in response to ATP, nigericin and MSU. These findings suggest that CZE effectively inhibited activation of the NLRP3 inflammasome.

Keywords: Chrysanthemum zawadskii; IL-1β; apoptosis-associated speck-like protein containing a caspase-recruitment domain oligomerization; bone marrow-derived macrophage; nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 inflammasome.

Grants and funding

Funding: The present study was supported by the Korea Research Institute of Bioscience and Biotechnology Research Initiative Program (grant no. KGM4571922) and the Korean Health Technology R&D Project funded by the Ministry of Health and Welfare (grant no. HI17C1238).