Journal: Journal of Translational Medicine

Article Title: Microbiota-derived tryptophan metabolites indole-3-lactic acid is associated with intestinal ischemia/reperfusion injury via positive regulation of YAP and Nrf2

doi: 10.1186/s12967-023-04109-3

Figure Lengend Snippet: ILA improved the survival rate and inhibited apoptosis of enterocytes under I/R and H/R injury. A Survival rate between sham, I/R and I/R + ILA groups (n = 16/group). B-C H&E staining ( B ) and quantification ( C ) of the histopathology changes of intestinal tissue sections (n = 3/group). D Relative serum LDH levels in mice under intestinal I/R injury (n = 4/group). E The effect of different ILA concentration on organoids’ cell viability under H/R injury (n = 5/group). F Representative images of morphological changes and PI staining in organoids. G Quantification of PI staining (n = 3/group). H Relative LDH levels in the organoids culture supernatant (n = 3–4/group). I AhR and CYP1A1 mRNA levels in intestinal tissues (n = 5/group). J YAP mRNA levels in organoids (n = 5/group). K Western blot for the protein expression of YAP in intestinal tissues (3 representative cases in each group). L YAP mRNA levels in intestinal tissues (n = 4/group). M YAP mRNA levels in organoids (n = 5/group). Results are presented as mean ± SEM. The statistical tests employed included: two-tailed log-rank test in ( A ), two-tailed student’s t-test in C – E , G – I , L and M and one-way ANOVA followed by the Tukey test for multiple comparisons in J . * p < 0.05, ** p < 0.01, **** p < 0 .0001

Article Snippet: For hypoxia-reoxygenation (H/R) experiments, organoids were precultured in ILA (0.5 mM), CH-223191 (10 μM, T2448, TargetMol) or VP (5 μM) for 24 h. It was then placed in a 95% N 2 -5% CO 2 humidity incubator for 12 h and further moved into a 95% O 2 -5% CO 2 incubator for 4 h. The organoids were randomized into five groups before being subjected to H/R experiments: (1) Control + DMSO; (2) H/R + DMSO; (3) H/R + ILA; (4) H/R + ILA + VP; (5) H/R + VP.

Techniques: Staining, Histopathology, Tissue, Concentration Assay, Organoids, Western Blot, Expressing, Two Tailed Test

Journal: Journal of Translational Medicine

Article Title: Microbiota-derived tryptophan metabolites indole-3-lactic acid is associated with intestinal ischemia/reperfusion injury via positive regulation of YAP and Nrf2

doi: 10.1186/s12967-023-04109-3

Figure Lengend Snippet: ILA protected intestinal organoids against H/R injury in vitro. A YAP mRNA expression in intestinal organoids (n = 6/group). B Representative images of protein levels in intestinal organoids (Four independent experiments). C - D YAP downstream gene Cyr61 ( C ), Ctgf ( D ) expression in intestinal organoids (n = 6/group). E ELISA analysis of the cytokine TNF-α and IL-6 in the supernatant medium (n = 4/group). F Relative LDH levels in the supernatant medium (n = 4–5/group). G Immunofluorescence of YAP and Ki67 in intestinal organoids frozen sections, TUNEL staining in intestinal organoids frozen sections. H - J Quantification of YAP protein expression ( H ), Ki67 + cells ( I ) and TUNEL + cells ( J ) in organoids (n = 3/group). Results are presented as mean ± SEM. The statistical tests employed included: one-way ANOVA followed by the Tukey test for multiple comparisons in A , C , D – F and H – J . * p < 0.05, ** p < 0.01, *** p < 0 .001, **** p < 0 .0001

Article Snippet: For hypoxia-reoxygenation (H/R) experiments, organoids were precultured in ILA (0.5 mM), CH-223191 (10 μM, T2448, TargetMol) or VP (5 μM) for 24 h. It was then placed in a 95% N 2 -5% CO 2 humidity incubator for 12 h and further moved into a 95% O 2 -5% CO 2 incubator for 4 h. The organoids were randomized into five groups before being subjected to H/R experiments: (1) Control + DMSO; (2) H/R + DMSO; (3) H/R + ILA; (4) H/R + ILA + VP; (5) H/R + VP.

Techniques: Organoids, In Vitro, Expressing, Enzyme-linked Immunosorbent Assay, Analysis, Immunofluorescence, TUNEL Assay, Staining

Journal: Journal of Translational Medicine

Article Title: Microbiota-derived tryptophan metabolites indole-3-lactic acid is associated with intestinal ischemia/reperfusion injury via positive regulation of YAP and Nrf2

doi: 10.1186/s12967-023-04109-3

Figure Lengend Snippet: ILA reduced excessive inflammation and oxidative stress in enterocytes both in vivo and in vitro. A - B Relative GSH level ( A ) and MDA level ( B ) in the serum (n = 4/group). C Representative images and quantification of the protein level of Nrf2 in intestinal tissue (3 representative cases in each group). D Relative LDH level in the serum (n = 4/group). E Relative gene expression of TNFα, IL-1β and IL-6 in intestinal tissue (n = 4/group). F - H Representative images and quantification of HE staining ( F ), immunohistochemical staining of Occludin ( G ) and ZO-1 ( H ) protein expression in the intestine tissues (n = 3–4/group). I Representative images and quantification of TUNEL + cells in intestinal organoids frozen sections. J - L Relative LDH level ( J ), GSH level ( K ) and MDA level ( L ) in the culture supernatants of organoids (n = 4/group). Results are presented as mean ± SEM. The statistical tests employed included: two-tailed student’s t-test in A – C , two-way ANOVA followed by the Tukey test for multiple comparisons in D – L . * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: For hypoxia-reoxygenation (H/R) experiments, organoids were precultured in ILA (0.5 mM), CH-223191 (10 μM, T2448, TargetMol) or VP (5 μM) for 24 h. It was then placed in a 95% N 2 -5% CO 2 humidity incubator for 12 h and further moved into a 95% O 2 -5% CO 2 incubator for 4 h. The organoids were randomized into five groups before being subjected to H/R experiments: (1) Control + DMSO; (2) H/R + DMSO; (3) H/R + ILA; (4) H/R + ILA + VP; (5) H/R + VP.

Techniques: In Vivo, In Vitro, Tissue, Gene Expression, H&E Stain, Immunohistochemical staining, Staining, Expressing, TUNEL Assay, Organoids, Two Tailed Test