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. 2018 Oct 15;9:1171. doi: 10.3389/fphar.2018.01171

FIGURE 2.

FIGURE 2

NOVO attenuated the expression of TRPV1. (A,B) Effects of NOVO on the expression of TRPV1 mRNA in IEC-6/TRPV1+ for 12 h (A) and 24 h (B). IEC-6/TRPV1+ cells were treated with 2.5–10 μM NOVO for 12 or 24 h, or 10 μM RR for 12 or 24 h, or 0.1 μM SB-705498 for 12 or 24 h, 25 μM CAP for 12 or 24 h, or 0.1% DMSO for 12 or 24 h. Shown are the mean relative mRNA expression levels normalized to untreated cells from three independent experiments performed in triplicate with standard deviations. ∗p < 0.05 vs. control group. (C) The expression of TRPV1 protein in non-transfected and transfected cells. (D) Effects of NOVO on mRNA expression of TRPV1 in jejunum, ileum and colon. Shown are the mean relative mRNA expression levels normalized to NC from three independent experiments performed in triplicate with standard deviations. ∗p < 0.05, statistic versus NC. Each value represents mean ± SD (n = 5). Rats were exposed to different concentrations of NOVO (5, 10, 25, and 50 μM), RR (10 μM) or 0.9% saline (NC) through oral administration (p.o.) twice per day for 14 days. Total RNA was obtained from the intestinal tissues extracted from treated rats. (E) Effects of NOVO on the protein expression of TRPV1 in jejunum, ileum and colon. Total membrane proteins were obtained from the intestinal tissues extracted from treated rats. (F) Effects of NOVO on the expression of TRPV1 protein in IEC-6/TRPV1+. (G) Subcellular distribution of TRPV1 in the transfected cell lines. IEC-6/TRPV1+ cells were fixed for immunofluorescence testing followed by pre-treatment with NOVO (10 μM), SB-705498 (0.1 μM), or 0.1% DMSO for 24 h and then labeled with anti-TRPV1 antibody (green) and DAPI (blue). Original magnification × 400. ∗p < 0.05, statistic versus control. Each value represents mean ± SD (n = 5).