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Published on: May 31, 2018
3,3'-Diindolylmethane suppresses the inflammatory response to lipopolysaccharide in murine macrophages
Han Jin Cho1, Mi Ra Seon, Yeo Myeong Lee
1Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University, Chuncheon 200-702, South Korea.
Abstract:
3,3'-Diindolylmethane (DIM), a major acid-condensation product of indole-3-carbinol, has been shown to have multiple anticancer effects in experimental models. Because recurrent or chronic inflammation has been implicated in the development of a variety of human cancers, this study examined the antiinflammatory effects of DIM and the underlying mechanisms using lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages. DIM significantly decreased the release of nitric oxide (NO), prostaglandin (PG)E2, tumor necrosis factor alpha, interleukin (IL)-6, and IL-1beta by RAW264.7 cells treated with LPS. DIM inhibited LPS-induced increases in protein levels of inducible NO synthase (iNOS), which were accompanied by decreased iNOS mRNA levels and transcriptional activity. The mRNA levels of phospholipase A2 decreased, whereas neither cyclooxygenases-2 protein nor transcript was altered by DIM. In addition, DIM suppressed LPS-induced nuclear factor-kappaB (NF-kappaB) transcriptional activity, NF-kappaB DNA-binding activity, translocation of p65 (RelA) to the nucleus, and degradation of inhibitor of kappaB alpha. Furthermore, DIM decreased LPS-induced transcriptional activity of activator protein (AP)-1, AP-1 DNA-binding activity, and phosphorylation of stress-activated protein kinase/Jun-N-terminal kinase and c-Jun. We demonstrate that DIM inhibits LPS-induced release of proinflammatory mediators in murine macrophages. Downregulation of NF-kappaB and AP-1 signaling may be one of the mechanisms by which DIM inhibits inflammatory responses.
Insights
3,3'-Diindolylmethane (DIM) significantly reduces inflammatory responses in macrophages by inhibiting key signaling pathways. This compound, derived from indole-3-carbinol, demonstrates potent anti-inflammatory effects with potential anticancer applications.
Area of Science:
- Immunology
- Molecular Biology
- Pharmacology
Background:
- Chronic inflammation is linked to various human cancers.
- 3,3'-Diindolylmethane (DIM), a metabolite of indole-3-carbinol, exhibits anticancer properties.
- Understanding DIM's anti-inflammatory mechanisms is crucial for cancer prevention and treatment.
Purpose of the Study:
- To investigate the anti-inflammatory effects of DIM in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages.
- To elucidate the molecular mechanisms underlying DIM's anti-inflammatory actions.
- To assess DIM's impact on key inflammatory mediators and signaling pathways.
Main Methods:
- RAW264.7 murine macrophages were stimulated with LPS.
- DIM's effect on the release of nitric oxide (NO), prostaglandin E2 (PGE2), and cytokines (TNF-α, IL-6, IL-1β) was measured.
- Changes in protein and mRNA levels of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and phospholipase A2 (PLA2) were analyzed.
- Nuclear factor-kappaB (NF-κB) and activator protein-1 (AP-1) signaling pathways were assessed, including DNA-binding activity, nuclear translocation, and phosphorylation of key proteins.
Main Results:
- DIM significantly reduced the release of NO, PGE2, TNF-α, IL-6, and IL-1β.
- DIM inhibited LPS-induced increases in iNOS protein and mRNA levels.
- DIM suppressed NF-κB and AP-1 transcriptional activity, DNA-binding, and related signaling events.
- DIM decreased PLA2 mRNA levels but did not affect COX-2 expression.
Conclusions:
- DIM effectively inhibits the release of pro-inflammatory mediators in LPS-stimulated macrophages.
- Downregulation of NF-κB and AP-1 signaling pathways is a key mechanism for DIM's anti-inflammatory effects.
- DIM shows promise as a therapeutic agent for inflammatory conditions and potentially as an anticancer agent.
