Title:5-methylthiopentyl Isothiocyanate, a Sulforaphane Analogue, Inhibits
Pro-inflammatory Cytokines by Regulating LPS/ATP-mediated NLRP3
Inflammasome Activation
Volume: 25
Issue: 5
Author(s): Su-Bin Choi, Ji-Hye Kim, Sehee Kwon, Na-Hyun Ahn, Joo-Hee Lee, Woong-Suk Yang, Cheorl-Ho Kim*Seung-Hoon Yang*
Affiliation:
- Department of Biological Sciences, Sungkyunkwan University and Samsung Advanced Institute of Health
Science and Technology, Suwon, 1649, Republic of Korea
- Department of Biomedical Engineering, College of Life Science and Biotechnology, Dongguk University, Seoul, 04620,
Republic of Korea
Keywords:
5-methylthiopentyl isothiocyanate, a sulforaphane analogue (berteroin), inflammation, NLRP3 inflammasome, primary bone marrow-derived macrophages (BMDMs), astrocyte, pro-inflammatory cytokine, ASC speck.
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.
