Multiple mechanistic anti-inflammatory activity of STW5-II on mouse intestinal organoids

Background Intestinal inflammation is a key player in the pathogenesis of many functional and organic gastrointestinal diseases. Inflammatory bowel diseases such as ulcerative colitis and Crohn´s disease are characterized by high grade inflammation, while irritable bowel syndrome is associated with low-grade inflammation.

Aim To establish organoids-based model of intestinal inflammation and to investigate the molecular mechanisms of the herbal medicinal product STW 5-II.

Methods Mouse intestinal organoids were cultured in Matrigel domes. Organoids were treated with increasing doses of STW 5-II for 24 hours followed by inflammation induction. A cocktail of pro-inflammatory cytokines (IFNγ, TNFα, IL-1β, IL-6) and bacterial components (lipopolysaccharide LPS-B5, flagellin) was used to induce intestinal inflammation. The exact mechanisms of STW 5-II were elaborated using microarrays analysis, qPCR, western blot, immunohistochemistry and immunofluorescence. We have investigated the expression of many pro-inflammatory genes such as STAT1, NFκB, NOS2, TNFα and IFNγ in addition to some anti-inflammatory mediators such as SIRT1 and IL10.

Results Microarrays analyses highlighted multiple inflammatory pathways that could be affected by STW 5-II. It decreases the expression of pro-inflammatory mediators such as STAT1, NFκB, NOS2, TNFα and IFNγ, while it upregulates the anti-inflammatory mediators such as SIRT1 and IL10. These findings were further confirmed by qPCR. Corresponding protein expression was confirmed by immunoblotting, IHC and immunofluorescence. STW 5-II was effective in a dose dependent manner in attenuating the cytokines-mediated inflammation within the organoids.

Conclusion STW 5-II shows very potent anti-inflammatory activity with multiple mechanistic targets. STW 5-II could be a promising therapy to alleviate intestinal inflammation associated with IBS, UC and Crohn´s disease.

Publikationsverlauf

Artikel online veröffentlicht:
08. September 2020