gr 113808  (Tocris)

Journal: Translational Pediatrics

Article Title: Chenodeoxycholic acid activates the TGR5/TRPA1-5-HT pathway to regulate intestinal motility in breastfed infants and mouse models

doi: 10.21037/tp-2025-100

Figure Lengend Snippet: Impact of serotonin pathway inhibition on CDCA-induced changes in intestinal motility as described in . After CDCA treatment (20 mg/kg via enema for 7 days), mice received inhibitors targeting different components of the serotonin signaling pathway. (A) Representative images of GI transit distance after carmine red gavage (30 min) in mice treated with: Control (PBS), CDCA only, Alosetron (5-HT3R antagonist, 1 mg/kg, blocking serotonin receptor signaling), GR113808 (5-HT4R antagonist, 1 mg/kg, blocking a different serotonin receptor subtype), or LX1606 (Tph1 inhibitor, 200 mg/kg, blocking serotonin synthesis). The red arrow as in . (D) Similar transit images for mice treated with Control, CDCA only, SBI-115 (TGR5 antagonist, 15 mg/kg, blocking the bile acid receptor upstream of serotonin signaling), or HC-030031 (TRPA1 inhibitor, 150 mg/kg, blocking another upstream component in the signaling pathway). The red arrow as in . (B,E) Representative images of fecal appearance from corresponding treatment groups. (C,F) Quantitative measurements of fecal water content, GI transit time, colonic transit distance (60 min post-carmine), and ELISA results for 5-HT and cAMP levels in colon tissue for all treatment groups. The experimental design systematically targets different points in the TGR5/TRPA1-5-HT signaling axis as detailed in the Methods section (2.8 and 2.9). Data were presented as mean ± SD. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001. n=5/group. 5-HT, 5-hydroxytryptamine (serotonin); 5-HT3R, 5-HT receptor 3; 5-HT4R, 5-HT receptor 4; cAMP, cyclic adenosine monophosphate; CDCA, chenodeoxycholic acid; ELISA, enzyme-linked immunosorbent assay; GI, gastrointestinal; PBS, phosphate-buffered saline; SD, standard deviation; Tph1, tryptophan hydroxylase 1; TRPA1, transient receptor potential A1.

Article Snippet: LX1606 (T171759, Aladdin; listed in ) was used as a Tph1 inhibitor , while alosetron (A125218, Aladdin; listed in ) and GR113808 (HY-103152, Mce; listed in ) served as 5-HT3R and 5-HT4R antagonists, respectively ( , ).

Techniques: Inhibition, Control, Blocking Assay, Enzyme-linked Immunosorbent Assay, Saline, Standard Deviation

Journal: Journal of Inflammation Research

Article Title: Early-Life Stress Induced by Neonatal Maternal Separation Leads to Intestinal 5-HT Accumulation and Causes Intestinal Dysfunction

doi: 10.2147/JIR.S488290

Figure Lengend Snippet: 5-HT promoted neurogenesis in the enteric nervous system. ( A ) The mRNA expression of 5-HT 3 , 5-HT 4 , and 5-HT 7 in the colon (n=6). ( B ) The protein expression of 5-HT 4 and GAPDH in the colon (n=3). ( C ) The mRNA expression of Tubb3, Sox2, Nestin, Olig2, and Gfap in the muscularis (n=6). ( D ) The protein expression of Tuj1, HUC/D, and GAPDH in the muscularis (n=3). ( E ) Immunofluorescence staining of HUC/D (green) and Tuj1 (red) in the LMMP (scale bar=100μm), and the number of HUC/D ( F ) and Tuj1 ( G ) positive cells was quantified. ( H ) Immunofluorescence staining of Tuj1 in the colon (Tuj1, red) and nucleic acid (Dapi, blue) (scale bar=100 μm), and ( I ) the number of Tuj1-positive cells was quantified. ( J ) Immunofluorescence staining of S100β in LMMP (S100β, red) and nucleic acid (Dapi, blue) (scale bar=100 μm), and ( K ) the number of S100β-positive cells was quantified. Data is presented as the mean ± SEM. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Subsequently, the cells were incubated with 5-HT (1 μM, H9523, Sigma‒Aldrich, Shanghai, China) along with the 5-HT 4 receptor antagonist GR113808 (10 nM, HY-103152, MCE, USA) or the Wnt pathway inhibitor IWP-2 (20 μM, HY-13912, MCE, USA) in serum-free media for 2 days.

Techniques: Expressing, Immunofluorescence, Staining

Journal: Journal of Inflammation Research

Article Title: Early-Life Stress Induced by Neonatal Maternal Separation Leads to Intestinal 5-HT Accumulation and Causes Intestinal Dysfunction

doi: 10.2147/JIR.S488290

Figure Lengend Snippet: 5-HT 4 mediates 5-HT activation of the Wnt signaling pathway to promote neurogenesis. ( A and B ) The protein expression levels of β-catenin, APC, Axin1, GSK-3β, and GAPDH in the muscularis muscle were determined by Western blotting (n=3). ( C ) Western blotting was used to determine the protein expression of β-catenin, and GAPDH in N2a cells (n=3). ( D ) The expression of Tubb3, Sox2, and Nestin in N2a cells was measured by real-time qPCR (n=6). ( E – H ) The protein expression of β-catenin, HUC/D, Tuj1, and GAPDH in N2a cells was examined by Western blotting, and the relative protein levels were normalized to those of GAPDH (n=3). Data is presented as the mean ± SEM. * P < 0.05; ** P < 0.01.

Article Snippet: Subsequently, the cells were incubated with 5-HT (1 μM, H9523, Sigma‒Aldrich, Shanghai, China) along with the 5-HT 4 receptor antagonist GR113808 (10 nM, HY-103152, MCE, USA) or the Wnt pathway inhibitor IWP-2 (20 μM, HY-13912, MCE, USA) in serum-free media for 2 days.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Journal of Inflammation Research

Article Title: Early-Life Stress Induced by Neonatal Maternal Separation Leads to Intestinal 5-HT Accumulation and Causes Intestinal Dysfunction

doi: 10.2147/JIR.S488290

Figure Lengend Snippet: Overview of the role of 5-HT signaling in ELS-induced intestinal dysfunction. Elevated serotonin levels in the gut due to early stress contribute to intestinal dysfunction in mice. These effects may occur through activation of the 5-HT 4 receptor, stimulation of the Wnt pathway, the promotion of neurogenesis, an increase in excitatory neurons, and subsequent impacts on intestinal motility.

Article Snippet: Subsequently, the cells were incubated with 5-HT (1 μM, H9523, Sigma‒Aldrich, Shanghai, China) along with the 5-HT 4 receptor antagonist GR113808 (10 nM, HY-103152, MCE, USA) or the Wnt pathway inhibitor IWP-2 (20 μM, HY-13912, MCE, USA) in serum-free media for 2 days.

Techniques: Activation Assay