Figure 6.

Biopsies from IBD patients incubated with NPY1R antagonist BIBP-3222 ex vivo depicted significant reduction in TNF. Human IBD biopsies obtained from the sigmoid colon were incubated overnight in RPMI media with or without NPY1R (BIBP) or NPY2R antagonist (BIIE; 100uM) at 5% CO₂, and supernatants were analyzed for TNF by multiplex ELISA using a human V-Plex pro-inflammatory MSD plate. UC and CD biopsy supernatants depicted enhanced release of (A) TNF, (B) IFN-γ, (C) IL-10, (D) IL-4, (E) IL-1β, and (F) IL-6 (n = 8-12 for control biopsies, n = 12-13 for UC, and n = 5-8 for CD). UC biopsies treated with BIBP showed significant downregulation of (G) TNF, whereas UC biopsies treated with BIBP and BIIE showed significant downregulation of IFN-γ (H). CD biopsies treated with BIBP or BIIE were ineffective in inhibiting TNF release (I); n = 10 for UC + BIBP, n = 5 for UC + BIIE, n = 6 for CD + BIBP, n = 4 for CD + BIIE. One-way ANOVA followed by Dunnett multiple comparison test was used to compare cytokine release in control, UC and CD biopsies. Paired t test was used to compare cytokine release between untreated and antagonist treated biopsies. Mean ± SEM, *P < .05, **P < .01, ***P < .001.