Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: HSD promoted Th17 cell development and worsened prostate inflammation in EAP mice. a Simple schematic diagram of the implementation plan of the experiment. b HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). c Tactile allodynia development in the 4 groups of mice ( n = 4). d The levels of IL-1β, TNF-α, IL-17A, and IFN-γ in the serum of the mice in the 4 groups ( n = 4). e Proportion of Th1 and Th17 cells of CD4 + T cells detected by flow cytometry in the splenic lymphocytes of vaccinated mice from those 4 groups ( n = 4). f The infiltration of Th17 cells in prostate tissues from mice was determined by immunofluorescence (white arrowheads, scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. BF breeding feed, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IFN-γ interferon-γ, IL-17A interleukin-17A, DAPI 4',6-diamidino-2-phenylindole, Th T helper

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: Transplantation of fecal microbiota from EAP mice with HSD elicited more severe prostate inflammation in psuedogerm-free mice. a Simple schematic diagram of the experimental workflow. b HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). c Development of tactile allodynia in mice belonging to the 4 groups ( n = 4). d The concentrations of IL-1β, TNF-α, IL-17A, and 5-HIAA in the serum of NOD mice in the 4 groups were measured ( n = 4). e Proportion of Th17 of CD4 + T cells, as determined by flow cytometry, among the splenic lymphocytes of immunized mice in the 4 groups ( n = 4). f The infiltration of Th17 cells in prostate tissues from mice was determined by immunofluorescence (white arrowheads, scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. NOD nonobese male diabetic/LtJ, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, FMT fecal microbiota transplantation, HE hematoxylin–eosin, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, DAPI 4',6-diamidino-2-phenylindole

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Transplantation Assay, Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: 5-HIAA supplementation ameliorated the symptoms of EAP caused by HSD. a The levels of 5-HIAA secreted into the serum of the mice were measured in 4 groups: Ctrl + NSD, Ctrl + HSD, EAP + NSD, and EAP + HSD groups ( n = 4). b Simple schematic diagram of the experimental workflow. c HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). d Tactile allodynia development in NOD mice in the 4 groups ( n = 4). e Secretion levels of IL-1β, TNF-α, and IL-17A in the serum of mice in the 5 groups ( n = 4). f Flow cytometry was used to determine the proportion of Th17 of CD4 + T cells among the splenic lymphocytes of immunized mice in the 5 groups ( n = 4). g The infiltration of Th17 cells in prostate tissues from mice was determined by immunofluorescence (white arrowheads, scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. NOD nonobese male diabetic/LtJ, BF breeding feed, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, HE hematoxylin–eosin, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, DAPI 4',6-diamidino-2-phenylindole

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: Inhibition of AHR abrogated the protective effects of 5-HIAA supplementation on EAP mice fed an HSD. a AHR levels in prostate tissues of mice among the Ctrl + NSD, Ctrl + HSD, EAP + NSD, EAP + HSD, and EAP + HSD + 5-HIAA groups ( n = 4). b A simple schematic diagram of the experimental workflow. c HE staining and inflammation scores of prostate tissues (scale bar = 100 μm, n = 4). d Tactile allodynia development in the mice from the Ctrl + NSD, Ctrl + HSD, EAP + NSD, EAP + HSD, EAP + HSD + 5-HIAA, and EAP + HSD + 5-HIAA + CH groups ( n = 4). e The levels of IL-1β, TNF-α, and IL-17A in the serum of the mice in the 6 groups ( n = 4). f Proportion of Th17 of CD4 + T cells, as determined by flow cytometry, among the splenic lymphocytes of immunized mice in the 6 groups (white arrowheads, n = 4). g The infiltration of Th17 cells in prostate tissues from mice was evaluated by immunofluorescence (scale bar = 100 μm, n = 4). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. NOD nonobese male diabetic/LtJ, BF breeding feed, EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, CH CH223191 (an AHR inhibitor), HE hematoxylin–eosin, IL-1β interleukin-1β, TNF-α tumor necrosis factor-α, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, AHR aryl hydrocarbon receptor, DAPI 4',6-diamidino-2-phenylindole

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: Inhibition, Staining, Flow Cytometry, Immunofluorescence

Journal: Military Medical Research

Article Title: High-salt-driven gut microbiota dysfunction aggravates prostatitis by promoting AHR/SGK1/FOXO1 axis-mediated Th17 cell differentiation

doi: 10.1186/s40779-025-00607-1

Figure Lengend Snippet: HSD promoted the differentiation of Th17 cells by activating SGK1/FOXO1 signaling pathway. a The in vitro Th17 cell differentiation ratio was determined by flow cytometry ( n = 3). b Th17 cell differentiation ratio was determined in an in vitro differentiation experiment using immunofluorescence (scale bar = 100 μm, n = 3). c SGK1, p-FOXO1, and FOXO1 levels in prostate tissues from Ctrl + NSD, Ctrl + HSD, EAP + NSD, EAP + HSD, and EAP + HSD + 5-HIAA mice ( n = 4). d AHR, SGK1, p-FOXO1, and FOXO1 levels in prostate tissues of mice in the EAP + HSD + 5-HIAA and EAP + HSD + 5-HIAA + CH groups ( n = 4). e AHR, SGK1, p-FOXO1, and FOXO1 levels in naïve CD4 + T cells of Ctrl, Sti, Sti + HSD, Sti + HSD + 5-HIAA, Sti + HSD + 5-HIAA + CH groups ( n = 3). f Transcriptional levels of Sgk1 in naïve CD4 + T cells of Ctrl, Sti, Sti + HSD, Sti + HSD + 5-HIAA, Sti + HSD + 5-HIAA + CH groups (n = 3). g For the ChIP-qPCR assay, 3 potential enhancer segments of the mouse promoter region of Sgk1 were created. h qPCR was used to assess the immunoprecipitated DNA by a ChIP assay ( n = 3). * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns non-significant. EAP experimental autoimmune prostatitis, HSD high-salt diet, NSD normal-salt diet, IL-17A interleukin-17A, 5-HIAA 5-hydroxyindole acetic acid, AHR aryl hydrocarbon receptor, SGK1 serum and glucocorticoid-regulated kinase 1, FOXO1 forkhead box protein O1, CH CH223191 (an AHR inhibitor), ChIP chromatin immunoprecipitation, qPCR quantitative polymerase chain reaction, DAPI 4’,6-diamidino-2-phenylindole, Sti stimulation with IL-6, IL-23, TGF-β, anti-IFN-γ, and anti-IL-4

Article Snippet: The following reagents and antibodies were used: CFA (#F5881, Sigma, USA); commercial enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ (#E-EL-M0048c), IL-1β (#E-EL-M0037), IL-17A (#E-EL-M0047c), tumor necrosis factor-α (TNF-α, #E-EL-M3063), and 5-HIAA (#E-EL-0075c) (Elabscience, China); anti-mouse CD4-fluorescein isothiocyanate (FITC, #553047), CD3e-allophycocyanin (APC, #561826), IFN-γ-phycoerythrin (PE, #562020), IL-17-PE (#559502), anti-human CD8-FITC (#555634), CD3-APC (#555342), IL-17-PE (#560436) (BD, USA); anti-CD3e (#BE0001-1), anti-CD28 (#BE0015-1), anti-IFN-γ (#BE0055), anti-IL-4 (#BE0045) (Bio X Cell, USA); IL-23 (#CS31), transforming growth factor-β1 (TGF-β1, #CA59), IL-6 (#CG39) (Novoprotein, China); anti-AHR (1:1000, #NB100-128, Novus, USA); anti-SGK1 (1:1000, #AF6789), anti-FOXO1 (1:1000, #AF6416), anti-p-FOXO1 (1:1000, #AF3417), anti-retinoic acid receptor-related orphan receptor gamma-t (RORγt; 1:1000, #DF3196), and anti-GAPDH (1:5000, #AF7021) (Affinity, USA).

Techniques: In Vitro, Cell Differentiation, Flow Cytometry, Immunofluorescence, ChIP-qPCR, Immunoprecipitation, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction

Journal: Journal of Orthopaedic Translation

Article Title: MATN3 delivered by exosome from synovial mesenchymal stem cells relieves knee osteoarthritis: Evidence from in vitro and in vivo studies

doi: 10.1016/j.jot.2023.06.003

Figure Lengend Snippet: MATN3 inhibits IL-1β-induced ECM degradation and autophagy defects in chondrocytes through interaction with IL-17A. A, RT-qPCR for IL-17A expression in chondrocytes. B, Western blot for IL-17A and MATN3 protein levels in chondrocytes. C, RT-qPCR for mRNA levels of IL-17A in the cartilage tissues of mice. D, Western blot for IL-17A and MATN3 protein levels in the cartilage tissue of mice. E, Co-IP experiment for the interaction between IL-17A and MATN3. F, Immunofluorescence for the co-localization of IL-17 and MATN3 in chondrocytes (bar ​= ​25 ​μm). G-H, ELISA for the IL-6 and TNF-α content in the supernatants of the chondrocytes. I, Western blot for expression of MMP-13, ADAMTS5, aggrecan, Collagen II, LC3II/LC3I, and Beclin1 in chondrocytes. ∗ indicates p ​< ​0.05 compared with the control group/sham; # indicates p ​< ​0.05 compared with the IL-17A ​+ ​oe-NC/OA ​+ ​oe-NC group; & indicates p ​< ​0.05 compared with the IL-17A ​+ ​sh-NC group. Cell experiments were performed in triplicate.

Article Snippet: Chondrocytes were treated with PBS (control), 10 ng/mL IL-1β, lentivirus of oe-MATN3, sh-MATN3, oe-NC, sh-NC, IL-17A (10 ng/mL; HY-P73174, MedChemExpress), DMSO (treated for 48 h), mTORC1 inhibitor rapamycin (rapa; 25 μM, S1039, Selleck, Shenzhen, China), an activator of PI3K/AKT pathway LY294002 (50 μM, S1105, Selleck), alone or in combination.

Techniques: Quantitative RT-PCR, Expressing, Western Blot, Co-Immunoprecipitation Assay, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Control

Journal: Journal of Orthopaedic Translation

Article Title: MATN3 delivered by exosome from synovial mesenchymal stem cells relieves knee osteoarthritis: Evidence from in vitro and in vivo studies

doi: 10.1016/j.jot.2023.06.003

Figure Lengend Snippet: MATN3 suppresses IL-1β-induced ECM degradation and autophagy defects in chondrocytes via the IL-17A/PI3K/AKT/mTOR axis. A Western blot for the protein levels of PI3K, AKT, and mTOR and phosphorylation levels of PI3K, AKT, and mTOR in cells. B, CCK-8 for the viability of chondrocytes. C-D, ELISA for IL-6 and TNF-α contents in the supernatants of chondrocytes. E, Western blot for the expression levels of MMP-13, ADAMTS5, aggrecan, Collagen II, LC3II/LC3I, and Beclin1 in chondrocytes. ∗ indicates p ​< ​0.05 compared with control/DMSO group; # indicates p ​< ​0.05 compared with the IL-1β + sh-NC/IL-1β + sh-IL-17A ​+ ​DMSO group; & indicates p ​< ​0.05 compared with the IL-1β ​+ ​oe-NC/IL-1β ​+ ​oe-MATN3 ​+ ​DMSO group. Cell experiments were performed in triplicate.

Article Snippet: Chondrocytes were treated with PBS (control), 10 ng/mL IL-1β, lentivirus of oe-MATN3, sh-MATN3, oe-NC, sh-NC, IL-17A (10 ng/mL; HY-P73174, MedChemExpress), DMSO (treated for 48 h), mTORC1 inhibitor rapamycin (rapa; 25 μM, S1039, Selleck, Shenzhen, China), an activator of PI3K/AKT pathway LY294002 (50 μM, S1105, Selleck), alone or in combination.

Techniques: Western Blot, Phospho-proteomics, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Expressing, Control

Journal: Allergologia et immunopathologia

Article Title: SIRT5 reduces the inflammatory response and barrier dysfunction in IL-17A-induced epidermal keratinocytes.

doi: 10.15586/aei.v51i1.675

Figure Lengend Snippet: Figure 1 IL-17A decreased SIRT5 expression. (A, B) The expression of SIRT5 by RT-qPCR and western blot assay. **P < 0.01, ***P < 0.001.

Article Snippet: To induce an inflammatory response, interleukin 17A (IL-17A, Absin, Shanghai, China) at concentrations of 10, 50, and 100 ng/ml was used to treat NHEKs for 24 h. To activate the ERK, activator tBHQivator t-butylhydroquinone (tBHQ, MedChemExpress)11 at a concentration of 50 μM was used to treat NHEKs for 24 h. abnormally low expression of SIRT5 might have a role in psoriasis (Figures 1A and 1B).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Allergologia et immunopathologia

Article Title: SIRT5 reduces the inflammatory response and barrier dysfunction in IL-17A-induced epidermal keratinocytes.

doi: 10.15586/aei.v51i1.675

Figure Lengend Snippet: Figure 2 SIRT5 overexpression decreased proliferation in IL-17A-induced keratinocytes. (A, B) The expression levels of SIRT5 by RT-qPCR and western blot assay. (C) The cell viability levels by CCK8 assay. (D) EdU staining analysis for cell proliferation. **P < 0.01, ***P < 0.001.

Article Snippet: To induce an inflammatory response, interleukin 17A (IL-17A, Absin, Shanghai, China) at concentrations of 10, 50, and 100 ng/ml was used to treat NHEKs for 24 h. To activate the ERK, activator tBHQivator t-butylhydroquinone (tBHQ, MedChemExpress)11 at a concentration of 50 μM was used to treat NHEKs for 24 h. abnormally low expression of SIRT5 might have a role in psoriasis (Figures 1A and 1B).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Staining

Journal: Allergologia et immunopathologia

Article Title: SIRT5 reduces the inflammatory response and barrier dysfunction in IL-17A-induced epidermal keratinocytes.

doi: 10.15586/aei.v51i1.675

Figure Lengend Snippet: Figure 3 SIRT5 overexpression decreased the levels of inflammatory factors in IL-17A-induced keratinocytes. (A) The levels of IL-8, IL-1β, and IL-6 by ELISA assay. (B) Western blot analysis for p-p65 levels. ***P < 0.001.

Article Snippet: To induce an inflammatory response, interleukin 17A (IL-17A, Absin, Shanghai, China) at concentrations of 10, 50, and 100 ng/ml was used to treat NHEKs for 24 h. To activate the ERK, activator tBHQivator t-butylhydroquinone (tBHQ, MedChemExpress)11 at a concentration of 50 μM was used to treat NHEKs for 24 h. abnormally low expression of SIRT5 might have a role in psoriasis (Figures 1A and 1B).

Techniques: Over Expression, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Redox Biology

Article Title: IDO1 improves postischemic neovascularization in aged mice by boosting endothelial NAD + de novo synthesis and curbing endothelial senescence

doi: 10.1016/j.redox.2025.103695

Figure Lengend Snippet: IL-17A/F represses ECs Ido1 transactivation and NAD + biosynthesis, and facilitates ECs senescence in the ischemic hindlimb muscle of young mice. A) Schematic diagram for testing methods was indicated. (B) Bar plot of the most prominent categories by KEGG pathway (left) and GO analysis (right). (C) Western blot analysis and quantification of IDO1 in hindlimb muscle ECs 7 days after HLI. PBS group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗ P < 0.01, ∗∗∗ P < 0.001. (D) RT-qPCR quantification of Ido1 in hindlimb muscle ECs 7 days after HLI. PBS group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗ P < 0.05. (E) Western blot analysis and quantification of IDO1 and P16 in hindlimb muscle ECs 7 days after HLI. Con group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗ P < 0.01, ∗∗∗∗ P < 0.0001, ns: not significant. (F–I) RT-qPCR quantification of Ido1 (F), p16 (G) , p21 (H) and Lmnb1 (I) in hindlimb muscle ECs 7 days after HLI. Con group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns: not significant. (J–L) ROS (J), NAD + (K) and NO (L) levels quantification in in hindlimb muscle ECs 7 days after HLI. Data of Con group in J was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns: not significant. (M) Venn diagram of IDO1 transcriptional factors between mouse and human. (N) Western blot analysis and quantification of P-CREB and total-CREB in hindlimb muscle ECs 7 days after HLI. Young group was set as 1. Unpaired two-tail t -test, n = 3 per group, data are mean ± SEM, ∗∗∗ P < 0.001. (O, P) Western blot analysis and quantification of P-CREB, total-CREB (T-CREB) (O), IDO1(P) and P16 (P) in hindlimb muscle ECs 7 days after HLI. Con group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns: not significant. (Q–T) RT-qPCR quantification of Ido1 (Q), p16 (R) , p21 (S) and Lmnb1 (T) in hindlimb muscle ECs 7 days after HLI. Con group was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ns: not significant. (U–W) ROS (U), NAD + (V) and NO (W) levels quantification in in hindlimb muscle ECs 7 days after HLI. Data of Con group in U was set as 1. One-way ANOVA with Tukey multiple comparison test, n = 3 per group, data are mean ± SEM, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns: not significant.

Article Snippet: Mouse IL-17A protein (Catalog #HY- P70753 ), mouse IL-17F protein (Catalog #HY- P72584 ), mouse IL-17A/F protein (Catalog #HY- P77707 ), mouse IDO1 protein (Catalog #HY- P72616 ), IgG (Catalog #HY-P99002), brodalumab (Catalog #HY-P9925, an anti-interleukin-17-receptor IgG2 monoclonal antibody), and 666–15 (Catalog #HY-101120, CREB phosphorylation inhibitor) were purchased from Med-ChemExpress.

Techniques: Western Blot, Comparison, Quantitative RT-PCR

Journal: Redox Biology

Article Title: IDO1 improves postischemic neovascularization in aged mice by boosting endothelial NAD + de novo synthesis and curbing endothelial senescence

doi: 10.1016/j.redox.2025.103695

Figure Lengend Snippet: IL-17A/F counteracts Endothelial Ido1 overexpression-induced neovascularization under conditions of lower limb ischemia. (A) Scheme showing the experiment for rescuing the IL-17A/F-treated young mice by endothelial Ido1 overexpression. (B and C) Representative laser Doppler images (B) of hindlimbs of mice and quantification (C) of relative perfusion recovery as the blood flow ratio of the left limb to that of the right limb in AAV-NC + PBS, AAV-NC + IL-17A/F, AAV- Ido1 + PBS and AAV- Ido1 + IL-17A/F mice before and after HLI at the indicated time points. The areas under the curves of relative perfusion are shown, Two-way ANOVA with Tukey multiple comparison test, n = 6 per group, data are mean ± SEM, ∗AAV-NC + IL-17A/F vs. AAV-NC + PBS; # AAV- Ido1 + IL-17A/F vs. AAV-NC+ IL-17A/F, #/ ∗ P < 0.05, ### P < 0.001, ∗∗∗∗/ #### P < 0.0001. (D) Representative micro-CT images of ischemic hindlimbs 21 days after HLI. Scale bar, 3 mm. (E) Quantification of micro-CT analysis of arterial vasculature in ischemic hindlimbs. Two-way ANOVA with Tukey multiple comparison test, n = 6 per group, data are mean ± SEM, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001, ns: not significant. (F) Representative whole-mounted images of ischemic gracilis 21 days after HLI. (G) Quantification of the diameter of pre-existed collateral arteries (6 points per artery). One-way ANOVA with Tukey multiple comparison test, n = 6 per group, data are mean ± SEM, ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001. (H) Immunofluorescence staining for CD31 in ischemic gastrocnemius muscles 21 days after HLI is shown. Scale bar, 200 μm. (I) Quantification of the CD31-positive area in muscle. One-way ANOVA with Tukey multiple comparison test, n = 6 per group, data are mean ± SEM, ∗∗∗∗ P < 0.0001.

Article Snippet: Mouse IL-17A protein (Catalog #HY- P70753 ), mouse IL-17F protein (Catalog #HY- P72584 ), mouse IL-17A/F protein (Catalog #HY- P77707 ), mouse IDO1 protein (Catalog #HY- P72616 ), IgG (Catalog #HY-P99002), brodalumab (Catalog #HY-P9925, an anti-interleukin-17-receptor IgG2 monoclonal antibody), and 666–15 (Catalog #HY-101120, CREB phosphorylation inhibitor) were purchased from Med-ChemExpress.

Techniques: Over Expression, Comparison, Micro-CT, Immunofluorescence, Staining, Muscles

Journal: Acta pharmacologica Sinica

Article Title: Pirfenidone ameliorates silica-induced lung inflammation and fibrosis in mice by inhibiting the secretion of interleukin-17A.

doi: 10.1038/s41401-021-00706-4

Figure Lengend Snippet: Fig. 3 Pirfenidone inhibited the secretion of IL-17A in early-stage silicosis models. ELISA was used to detect the protein levels of IL-17A in serum (a) and BALF (b). The mRNA and protein levels of IL-17A in lung tissues from different groups were measured by qPCR (c), immunohistochemistry (d), and Western blot analysis (e). All data were presented as mean ± SD; n = 10 for each group; ***P < 0.001, ****P < 0.0001, ns (no statistical significance).

Article Snippet: IL-17A neutralization By intraperitoneal injection, 100 μg of anti-mouse IL-17A antibody (M00421-2, BOSTER, Wuhan, China) or isotype control IgG (ab18443, abcam, Cambridge, UK) was given to silica-exposed mice weekly for 4 weeks, starting 1 day after silica exposure.

Techniques: Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Western Blot

Journal: Acta pharmacologica Sinica

Article Title: Pirfenidone ameliorates silica-induced lung inflammation and fibrosis in mice by inhibiting the secretion of interleukin-17A.

doi: 10.1038/s41401-021-00706-4

Figure Lengend Snippet: Fig. 6 Pirfenidone diminished the production of IL-17A in advanced silicosis models. ELISA was performed to measure the protein expression of IL-17A in serum (a) and BALF (b) from different groups. The mRNA and protein levels of IL-17A in lung tissues were measured by qPCR (c), immunohistochemistry (d) and Western blot analysis (e), respectively. All data were presented as mean ± SD; n = 10 for each group; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: IL-17A neutralization By intraperitoneal injection, 100 μg of anti-mouse IL-17A antibody (M00421-2, BOSTER, Wuhan, China) or isotype control IgG (ab18443, abcam, Cambridge, UK) was given to silica-exposed mice weekly for 4 weeks, starting 1 day after silica exposure.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Immunohistochemistry, Western Blot