2. Academic Validation
  3. Melampodium divaricatum (Rich.) DC. extracts exert anti-inflammatory effects via inhibition of the NLRP3 inflammasome

Melampodium divaricatum (Rich.) DC. extracts exert anti-inflammatory effects via inhibition of the NLRP3 inflammasome

  • J Ethnopharmacol. 2026 Oct 28:369:121903. doi: 10.1016/j.jep.2026.121903.
Seongjong Lee 1,  Seoyeon Jang 2,  Ja-Gyeong Song 3,  Jaehyeok Lee 4,  Indiana M Coronado 5,  Hyung Won Ryu 6,  Jung Hee Kim 7,  Dong-Keun Yi 8,  Soo-Yong Kim 9,  Sangho Choi 10,  Hyungseok Seo 11,  Man S Kim 12,  Jin-Hyub Paik 13,  Jongmin Ahn 14,  Yoonsung Lee 15,  Yong Hwan Park 16
Affiliations

Affiliations

  • 1 BK21 R&E Initiative for Advanced Precision Medicine, Suwon, 16499, Republic of Korea; Department of Microbiology, Ajou University School of Medicine, Suwon, 16499, Republic of Korea; Department of Biomedical Sciences, Graduate School of Ajou University, Suwon, Republic of Korea. Electronic address: [email protected].
  • 2 Department of Biomedical Science and Technology, Kyung Hee University, Seoul, 05278, Republic of Korea; Clinical Research Institute, Kyung Hee University Hospital at Gangdong, School of Medicine, Kyung Hee University, Seoul, 05278, Republic of Korea. Electronic address: [email protected].
  • 3 Natural Product Research Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju-si, Chungcheongbuk-do, 28116, Republic of Korea. Electronic address: [email protected].
  • 4 BK21 R&E Initiative for Advanced Precision Medicine, Suwon, 16499, Republic of Korea; Department of Microbiology, Ajou University School of Medicine, Suwon, 16499, Republic of Korea; Department of Biomedical Sciences, Graduate School of Ajou University, Suwon, Republic of Korea. Electronic address: [email protected].
  • 5 Herbarium of National Autonomous University of Nicaragua at Leon, Leon, 21000, Nicaragua. Electronic address: [email protected].
  • 6 Natural Product Research Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju-si, Chungcheongbuk-do, 28116, Republic of Korea. Electronic address: [email protected].
  • 7 Natural Product Research Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju-si, Chungcheongbuk-do, 28116, Republic of Korea. Electronic address: [email protected].
  • 8 International Biological Material Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 34141, Republic of Korea. Electronic address: [email protected].
  • 9 International Biological Material Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 34141, Republic of Korea. Electronic address: [email protected].
  • 10 International Biological Material Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 34141, Republic of Korea. Electronic address: [email protected].
  • 11 Department of Anesthesiology and Pain Medicine, Kyung Hee University Hospital at Gangdong, College of Medicine, Kyung Hee University, Seoul, 05278, Republic of Korea. Electronic address: [email protected].
  • 12 Clinical Research Institute, Kyung Hee University Hospital at Gangdong, School of Medicine, Kyung Hee University, Seoul, 05278, Republic of Korea; Center for Space Biomedical Sciences, NEXUS Institute, Kyung Hee University, Seoul, 05278, Republic of Korea. Electronic address: [email protected].
  • 13 International Biological Material Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 34141, Republic of Korea. Electronic address: [email protected].
  • 14 Natural Product Research Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju-si, Chungcheongbuk-do, 28116, Republic of Korea. Electronic address: [email protected].
  • 15 Department of Biomedical Science and Technology, Kyung Hee University, Seoul, 05278, Republic of Korea; Clinical Research Institute, Kyung Hee University Hospital at Gangdong, School of Medicine, Kyung Hee University, Seoul, 05278, Republic of Korea; Center for Space Biomedical Sciences, NEXUS Institute, Kyung Hee University, Seoul, 05278, Republic of Korea. Electronic address: [email protected].
  • 16 BK21 R&E Initiative for Advanced Precision Medicine, Suwon, 16499, Republic of Korea; Department of Microbiology, Ajou University School of Medicine, Suwon, 16499, Republic of Korea; Department of Biomedical Sciences, Graduate School of Ajou University, Suwon, Republic of Korea. Electronic address: [email protected].
PMID: 42202918 DOI: 10.1016/j.jep.2026.121903
Abstract

Ethnopharmacological relevance: Melampodium divaricatum (Rich.) DC. (MelD), commonly known as butter daisy, is a medicinal plant widely distributed in Central and South America. In traditional medicine, particularly in Mexico and Other parts of Central America, decoctions prepared from the aerial parts of MelD have been widely used to treat inflammation-associated ailments, including dysentery, fever, infections, and pain. Despite its long-standing ethnomedicinal use, the molecular and immunological mechanisms underlying its anti-inflammatory effects, particularly in the context of innate immune regulation, remain largely unexplored.

Aim of the study: Based on the ethnomedicinal use of MelD in Nicaragua for the treatment of inflammation-related conditions such as fever and pain, this study aimed to investigate whether extracts of MelD modulate the activation of the NLRP3 inflammasome, a central regulator of innate immune responses. In addition, we sought to elucidate the underlying molecular mechanisms and to characterize the major bioactive constituents associated with these anti-inflammatory effects.

Materials and methods: Methanolic extracts were prepared from the aerial parts of MelD collected in Nicaragua. Cytotoxicity was evaluated using the EZ-CYTOX cell viability assay. The effects of the extract on NLRP3 inflammasome activation were examined in lipopolysaccharide (LPS)-primed human THP-1 macrophages and murine macrophage cell lines. Inflammasome activation was induced by ATP or nigericin stimulation. Interleukin-1β (IL-1β) secretion was quantified by enzyme-linked immunosorbent assay (ELISA), while ASC speck formation and oligomerization were assessed by immunofluorescence microscopy and chemical cross-linking, respectively. NLRP3 ATPase activity was measured using the ADP-Glo™ Max assay. The in vivo anti-inflammatory activity of the extract was further evaluated using chemical- and tail-fin injury-induced zebrafish inflammation model. Chemical profiling of the extract was performed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS).

Results: The MelD extract significantly inhibited NLRP3 inflammasome activation and subsequent IL-1β secretion in a concentration-dependent manner without inducing cytotoxicity. Mechanistic analyses revealed that the extract suppressed ASC speck formation and ASC oligomerization, indicating an impairment of inflammasome assembly. Notably, treatment with the extract did not affect the expression levels of NLRP3 or pro-IL-1β during the priming phase, suggesting preferential interference with the activation step of the inflammasome. In agreement with the in vitro findings, the extract markedly reduced inflammatory responses, including neutrophil and macrophage recruitment, in the zebrafish inflammation model. UPLC-QTOF/MS analysis identified flavonoid C-glycosides as major constituents of the extract, implicating these compounds as potential contributors to the observed anti-inflammatory activity.

Conclusions: This study provides experimental evidence that MelD extract exerts anti-inflammatory effects by targeting the assembly and activation of the NLRP3 inflammasome. These findings offer a pharmacological rationale for the traditional use of MelD in the management of fever and pain and highlight its potential as a natural source of bioactive compounds for the development of therapeutics against NLRP3-driven inflammatory disorders.

Keywords

Anti-inflammatory; IL-1β; Melampodium divaricatum; NACHT domain; NLRP3 inflammasome; Plant extract.

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