5-Methylthiopentyl Isothiocyanate, a Sulforaphane Analogue, Inhibits Pro-inflammatory Cytokines by Regulating LPS/ATP-mediated NLRP3 Inflammasome Activation

Su-Bin Choi; Ji-Hye Kim; Sehee Kwon; Na-Hyun Ahn; Joo-Hee Lee; Woong-Suk Yang; Cheorl-Ho Kim; Seung-Hoon Yang

doi:10.2174/1389201024666230824093927

5-Methylthiopentyl Isothiocyanate, a Sulforaphane Analogue, Inhibits Pro-inflammatory Cytokines by Regulating LPS/ATP-mediated NLRP3 Inflammasome Activation

Curr Pharm Biotechnol. 2023 Aug 24. doi: 10.2174/1389201024666230824093927. Online ahead of print.

Authors

Su-Bin Choi¹, Ji-Hye Kim¹, Sehee Kwon¹, Na-Hyun Ahn¹, Joo-Hee Lee¹, Woong-Suk Yang², Cheorl-Ho Kim³, Seung-Hoon Yang¹

Affiliations

¹ Department of Biomedical Engineering, College of Life Science and Biotechnology, Dongguk University, Seoul 04620, Republic of Korea.
² National Institute for Nanomaterials Technology (NINT), POSTECH, 77, Pohang 37673, Republic of Korea.
³ Department of Biological Sciences, Sungkyunkwan University and Samsung Advanced Institute of Health Science and Technology, Suwon 1649, Republic of Korea.

PMID: 37622701
DOI: 10.2174/1389201024666230824093927

Abstract

Background: Pro-inflammatory cytokines secreted from activated macrophages and astrocytes are crucial mediators of inflammation for host defense. Among them, the secretion of IL-1β, a major pro-inflammatory cytokine, is especially mediated by the activation of NLRP3 inflammasome. Pro-IL-1β, which is produced in response to the invaded pathogens, such as LPS, is cleaved and matured in the NLRP3 inflammasome by the recognition of ATP. Excessively activated IL-1β induces other immune cells, resulting in the up-regulation of inflammation. Therefore, regulation of NLRP3 inflammasome can be a good strategy for alleviating inflammation.

Objective: Our study aimed to examine whether 5-methylthiopentyl isothiocyanate, a sulforaphane analogue (berteroin), has an anti-inflammatory effect on the NLRP3 inflammasome activation induced by LPS and ATP.

Methods: Primary bone marrow-derived macrophages (BMDMs) and astrocytes were stimulated by LPS and ATP with the treatment of 5-methylthiopentyl isothiocyanate, a sulforaphane analogue. The secretion of pro-inflammatory cytokines was measured by ELISA, and the expression level of NLRP3 inflammasome-associated proteins was detected by western blot. The association of NLRP3 inflammasome was assessed by co-immunoprecipitation, and the formation of ASC specks was evaluated by fluorescent microscope.

Results: 5-Methylthiopentyl isothiocyanate, a sulforaphane analogue (berteroin), decreased the release of pro-inflammatory cytokines, IL-1β, and IL-6 in the BMDMs. Berteroin notably prevented the formation of both NLRP3 inflammasome and ASC specks, which reduced the secretion of IL-1β. Additionally, berteroin reduced the IL-1β secretion and cleaved IL-1β expression in the primary astrocytes.

Discussion and conclusion: These results indicated the anti-inflammatory effects of 5-methylthiopentyl isothiocyanate (berteroin) by regulating NLRP3 inflammasome activation, suggesting that berteroin could be the potential natural drug candidate for the regulation of inflammation.

Keywords: 5-Methylthiopentyl isothiocyanate; ASC spec; NLRP3 inflammasome; astrocyte; primary bone marrow-derived macropahges (BMDMs); pro-inflammatory cytokine; sulforaphane analogue (berteroin).