Ethanol extract of <em>Chrysanthemum zawadskii</em> inhibits the NLRP3 inflammasome by suppressing ASC oligomerization in macrophages

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

doi:10.3892/etm.2023.11827

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

Authors:
- Ah-Ra Jang
- Ha-Nul Lee
- Jung Joo Hong
- Young-Min Kim
- Jong-Hwan Park
View Affiliations

Affiliations: Laboratory of Animal Medicine, College of Veterinary Medicine and Animal Medical Institute, Chonnam National University, Gwangju 61186, Republic of Korea, Department of Food Science & Technology and Brain Korea 21 Plus Program, Chonnam National University, Gwangju 61186, Republic of Korea, National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, Chungcheongbuk 28116, Republic of Korea, Department of Food Science & Technology and Brain Korea 21 Plus Program, Chonnam National University, Gwangju 61186, Republic of Korea, Laboratory of Animal Medicine, College of Veterinary Medicine and Animal Medical Institute, Chonnam National University, Gwangju 61186, Republic of Korea
Published online on: February 7, 2023 https://doi.org/10.3892/etm.2023.11827
Article Number: 128
Copyright: © Jang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

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.

View Figures

View References

1	Latz E, Xiao TS and Stutz A: Activation and regulation of the inflammasomes. Nat Rev Immunol. 13:397–411. 2013.PubMed/NCBI View Article : Google Scholar
2	Paik S, Kim JK, Silwal P, Sasakawa C and Jo EK: An update on the regulatory mechanisms of NLRP3 inflammasome activation. Cell Mol Immunol. 18:1141–1160. 2021.PubMed/NCBI View Article : Google Scholar
3	Kaneko N, Kurata M, Yamamoto T, Morikawa S and Masumoto J: The role of interleukin-1 in general pathology. Inflamm Regen. 39(12)2019.PubMed/NCBI View Article : Google Scholar
4	Mangan MSJ, Olhava EJ, Roush WR, Seidel HM, Glick GD and Latz E: Targeting the NLRP3 inflammasome in inflammatory diseases. Nat Rev Drug Discov. 17:588–606. 2018.PubMed/NCBI View Article : Google Scholar
5	Daniels MJD, Rivers-Auty J, Schilling T, Spencer NG, Watremez W, Fasolino V, Booth SJ, White CS, Baldwin AG, Freeman S, et al: Fenamate NSAIDs inhibit the NLRP3 inflammasome and protect against Alzheimer's disease in rodent models. Nat Commun. 7(12504)2016.PubMed/NCBI View Article : Google Scholar
6	Dixit VD: Nlrp3 inflammasome activation in type 2 diabetes: Is it clinically relevant? Diabetes. 62:22–24. 2013.PubMed/NCBI View Article : Google Scholar
7	Fusco R, Siracusa R, Genovese T, Cuzzocrea S and Di Paola R: Focus on the role of NLRP3 inflammasome in diseases. Int J Mol Sci. 21(4223)2020.PubMed/NCBI View Article : Google Scholar
8	Kanneganti TD: Inflammatory bowel disease and the NLRP3 inflammasome. N Engl J Med. 377:694–696. 2017.PubMed/NCBI View Article : Google Scholar
9	Guo C, Fu R, Wang S, Huang Y, Li X, Zhou M, Zhao J and Yang N: NLRP3 inflammasome activation contributes to the pathogenesis of rheumatoid arthritis. Clin Exp Immunol. 194:231–243. 2018.PubMed/NCBI View Article : Google Scholar
10	Sharma D and Kanneganti TD: The cell biology of inflammasomes: Mechanisms of inflammasome activation and regulation. J Cell Biol. 213:617–629. 2016.PubMed/NCBI View Article : Google Scholar
11	Coll RC, Robertson AA, Chae JJ, Higgins SC, Muñoz-Planillo R, Inserra MC, Vetter I, Dungan LS, Monks BG, Stutz A, et al: A small-molecule inhibitor of the NLRP3 inflammasome for the treatment of inflammatory diseases. Nat Med. 21:248–255. 2015.PubMed/NCBI View Article : Google Scholar
12	Cragg GM and Newman DJ: Nature products: A contrinuing source of novel drug leads. Biochim Biophys Acta. 1830:3670–3695. 2013.PubMed/NCBI View Article : Google Scholar
13	Kim AR, Kim HS, Kim DK, Lee JH, Yoo YH, Kim JY, Park SK, Nam ST, Kim HW, Park YH, et al: The extract of Chrysanthemum zawadskii var. latilobum ameliorates collagen-induced arthritis in mice. Evid Based Complement Alternat Med. 2016(3915013)2016.PubMed/NCBI View Article : Google Scholar
14	Gu DR, Hwang JK, Erkhembaatar M, Kwon KB, Kim MS, Lee YR and Lee SH: Inhibitory effect of Chrysanthemum zawadskii Herbich var. latilobum kitamura extract on RANKL-induced osteoclast differentiation. Evid Based Complement Alternat Med. 2013(509482)2013.PubMed/NCBI View Article : Google Scholar
15	Huang WY, Jeong I, Han BK, Kim MJ, Hong J, Ahn SII, Heo W, Pan JH, Kim JK, Shin EC and Kim YJ: Chrysanthemum zawadskii Herbich var. latilobum (Maxim.) Kitamura water extract prevents BALB/c mice lung injury from particulate matter 10 toxicity. Food Agric Immunol. 33:252–263. 2022.
16	Kim Y, Han J, Sung J, Sung M, Choi Y, Jeong HS and Lee J: Anti-inflammatory activity of Chrysanthemum zawadskii var. latilobum leaf extract through haem oxygenase-1 induction. J Funct Foods. 4:474–479. 2012.
17	Cho BO, Shin JY, Kang HJ, Park JH, Hao S, Wang F and Jang SI: Anti-inflammatory effect of Chrysanthemum zawadskii, peppermint, Glycyrrhiza glabra herbal mixture in lipopolysaccharide-stimulated RAW264.7 macrophages. Mol Med Rep. 24(532)2021.PubMed/NCBI View Article : Google Scholar
18	Kim KY, Oh TW, Yang HJ, Kim YW, Ma JY and Park KI: Ethanol extract of Chrysanthemum zawadskii Herbich induces autophagy and apoptosis in mouse colon cancer cells through the regulation of reactive oxygen species. BMC Complement Altern Med. 19(274)2019.PubMed/NCBI View Article : Google Scholar
19	Celada A, Gray PW, Rinderknecht E and Schreiber RD: Evidence for a gamma-interferon receptor that regulates macrophage tumoricidal activity. J Exp Med. 160:55–74. 1984.PubMed/NCBI View Article : Google Scholar
20	Rao X, Huang X, Zhou Z and Lin X: An improvement of the 2ˆ(-delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 3:71–85. 2013.PubMed/NCBI
21	Kelley N, Jeltema D, Duan Y and He Y: The NLRP3 inflammasome: An overview of mechanisms of activation and regulation. Int J Mol Sci. 20(3328)2019.PubMed/NCBI View Article : Google Scholar
22	Bryan NB, Dorfleutner A, Rojanasakul Y and Stehlik C: Activation of inflammasomes requires intracellular redistribution of the apoptotic speck-like protein containing a caspase recruitment domain. J Immunol. 182:3173–3182. 2009.PubMed/NCBI View Article : Google Scholar
23	Hara H, Tsuchiya K, Kawamura I, Fang R, Hernandez-Cuellar E, Shen Y, Mizuguchi J, Schweighoffer E, Tybulewicz V and Mitsuyama M: Phosphorylation of the adaptor ASC acts as a molecular switch that controls the formation of speck-like aggregates and inflammasome activity. Nat Immunol. 14:1247–1255. 2013.PubMed/NCBI View Article : Google Scholar
24	Zheng D, Liwinski T and Elinav E: Inflammasome activation and regulation: Toward a better understanding of complex mechanisms. Cell Discov. 6(36)2020.PubMed/NCBI View Article : Google Scholar
25	Hong S, Joo T and Jhoo JW: Antioxidant and anti-inflammatory activities of 3,5-dicaffeoylquinic acid isolated from Ligularia fischeri leaves. Food Sci Biotechnol. 24:257–263. 2015.
26	Hwang SJ, Kim YW, Park Y, Lee HJ and Kim KW: Anti-inflammatory effects of chlorogenic acid in lipopolysaccharide-stimulated RAW 264.7 cells. Inflamm Res. 63:81–90. 2014.PubMed/NCBI View Article : Google Scholar
27	Kim B, Lee JH, Seo MJ, Eom SH and Kim W: Linarin down-regulates phagocytosis, pro-inflammatory cytokine production, and activation marker expression in RAW264.7 macrophages. Food Sci Biotechnol. 25:1437–1442. 2016.PubMed/NCBI View Article : Google Scholar
28	Tőzsér J and Benkő S: Natural compounds as regulators of NLRP3 inflammasome-mediated IL-1β production. Mediators Inflamm. 2016(5460302)2016.PubMed/NCBI View Article : Google Scholar
29	Kim B and Kim HS: Chrysanthemum zawadskii extract activates peroxisome proliferator-activated receptor-α and has an anti-inflammatory activity: Potential interest for the skin barrier function. Korean J Chem Eng. 31:1831–1838. 2014.
30	Yu SH, Sun X, Kim MK, Akther M, Han JH, Kim TY, Jiang J, Kang TB and Lee KH: Chrysanthemum indicum extract inhibits NLRP3 and AIM2 inflammasome activation via regulating ASC phosphorylation. J Ethnopharmacol. 239(111917)2019.PubMed/NCBI View Article : Google Scholar
31	Lou L, Zhou J, Liu Y, Wei YI, Zhao J, Deng J, Dong B, Zhu L, Wu A, Yang Y and Chai L: Chlorogenic acid induces apoptosis to inhibit inflammatory proliferation of IL-6-induced fibroblast-like synoviocytes through modulating the activation of JAK/STAT and NF-κB signaling pathways. Exp Ther Med. 11:2054–2060. 2016.PubMed/NCBI View Article : Google Scholar
32	Shi A, Shi H, Wang Y, Liu X, Cheng Y, Li H, Zhao H, Wang S and Dong L: Activation of Nrf2 pathway and inhibition of NLRP3 inflammasome activation contribute to the protective effect of chlorogenic acid on acute liver injury. Int Immunopharmacol. 54:125–130. 2018.PubMed/NCBI View Article : Google Scholar
33	Han X, Wu YC, Meng M, Sun QS, Gao SM and Sun H: Linarin prevents LPS-induced acute lung injury by suppressing oxidative stress and inflammation via inhibition of TXNIP/NLRP3 and NF-κB pathways. Int J Mol Med. 42:1460–1472. 2018.PubMed/NCBI View Article : Google Scholar
34	Kwak SB, Koppula S, In EJ, Sun X, Kim YK, Kim MK, Lee KH and Kang TB: Artemisia extract suppresses NLRP3 and AIM2 inflammasome activation by inhibition of ASC phosphorylation. Mediators Inflamm. 2018(6054069)2018.PubMed/NCBI View Article : Google Scholar
35	Yang Y, Wang H, Kouadir M, Song H and Shi F: Recent advances in the mechanisms of NLRP3 inflammasome activation and its inhibitors. Cell Death Dis. 10(128)2019.PubMed/NCBI View Article : Google Scholar
36	Qu Y, Misaghi S, Newton K, Maltzman A, Izrael-Tomasevic A, Arnott D and Dixit VM: NLRP3 recruitment by NLRC4 during Salmonella infection. J Exp Med. 213:877–885. 2016.PubMed/NCBI View Article : Google Scholar
37	Jiang Y, Lin Y, Hu YJ, Song XJ, Pan HH and Zhang HJ: Caffeoylquinic acid derivatives rich extract from Gnaphalium pensylvanicum willd. Ameliorates hyperuricemia and acute gouty arthritis in animal model. BMC Complement Altern Med. 17(320)2017.PubMed/NCBI View Article : Google Scholar