bleomycin hydrochloride  (MedChemExpress)

Journal: iScience

Article Title: RSK1-SRF signaling axis drives fibroblast activation and pulmonary fibrosis: Genetic causality and therapeutic targeting

doi: 10.1016/j.isci.2026.115495

Figure Lengend Snippet: RSK1 is a causal risk factor for IPF and is upregulated in fibrotic lungs (A–D) Mendelian randomization (MR) analysis evaluating the causal association between RSK1 expression quantitative trait loci (eQTLs) and IPF risk, using the inverse variance weighted (IVW), MR Egger, weighted median, and weighted mode methods. (B and C) Scatter plot and weight plot for the IVW method, respectively. (D) Association strength between RSK1 eQTLs and IPF. (E) Boxplot of RPS6KA1 expression in lung tissues from IPF patients and healthy controls ( GSE70866 ; ∗ p < 0.05, as indicated). (F) Immunoblot analysis of phosphorylated RSK1 ( p -RSK1) and total RSK1 in mouse lungs on days 7 and 14 after bleomycin (BLM) challenge. (G) Densitometric quantification of (F); β-actin serves as the loading control. (H) Representative immunohistochemistry (IHC) staining images of p -RSK1 in mouse lungs during fibrosis progression. Scale bars, 100 μm. (I) Quantification of p -RSK1 IHC staining shown as positive area (%). (J and K) Representative IHC staining images of (J) RSK1 and (K) p -RSK1 in human IPF and normal lung tissues. Scale bars, 100 μm. (L) Immunofluorescence co-staining of p -RSK1 (green) with F4/80, E-cadherin (E-Cad), and fibronectin (FN) (red) in saline- and BLM-treated mouse lungs; nuclei are stained with DAPI (blue). Scale bars, 100 μm. Data are presented as the mean ± SD. For tissue-based assays, n denotes independent animals ( n = 6 per group). Image quantification was performed as described in ; technical sampling was not counted toward n . Statistical tests are described in . ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; # p ≥ 0.05.

Article Snippet: Bleomycin hydrochloride , MedChemExpress , Cat#HY-17565A.

Techniques: Expressing, Western Blot, Control, Immunohistochemistry, Immunofluorescence, Staining, Saline, Sampling

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: FoxM1 is highly expressed in fibroblasts of fibrotic lung tissues. (A, B) qPCR (n = 9) and Western blot (n = 6) analysis of the expression of FoxM1 in normal and IPF lung tissues. ∗ P < 0.05. (C) qPCR analysis of the mRNA levels of FoxM1 in the lung tissues from BLM-treated mice. n = 3, ∗ P < 0.05. (D) Western blot analysis of the protein levels of FoxM1, CTHRC1, α-SMA, and Collagen I in the lung tissues from bleomycin (BLM)-treated mice. n = 3, ∗ P < 0.05. (E) The Pearson's correlation analysis of COL1A1 expression with FoxM1 expression based on the RNA-seq results of GSE24206 from GEO database. (F) The Pearson's correlation analysis of Ashcroft score with FoxM1 expression in the lung tissues from BLM-treated mice. (G) Representative images of co-immunostaining for α-SMA and FoxM1 in IPF lung tissues. White arrows indicate double-positive cells. (H) Representative images of co-immunostaining for α-SMA and FoxM1 in the lung tissues from BLM-treated mice. White arrows indicate double-positive cells. (I) Western blot analysis of FoxM1 expression in pulmonary fibroblasts isolated from mice subjected to BLM treatment. n = 3, ∗ P < 0.05. (J) Western blot analysis of FoxM1, CTHRC1, α-SMA, and Collagen I expression in pulmonary fibroblasts treated with TGF-β1. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (A, B, I) and one-way ANOVA with Tukey's post-hoc test (C, D, J) were used for statistical analysis.

Article Snippet: For the induction of pulmonary fibrosis, a single dose of bleomycin (BLM, Nippon Kayaku, Tokyo, Japan) at 5 mg/kg, dissolved in 50 μl of sterile saline, was administered intratracheally to the treatment group.

Techniques: Western Blot, Expressing, RNA Sequencing, Immunostaining, Isolation

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Enhanced FoxM1 expression is responsible for the resistance of fibroblasts to FasL-mediated apoptosis . (A) Caspase 3 activity analysis in pulmonary fibroblasts isolated from bleomycin (BLM)-treated mice following FasL treatment. (B) Cell viability assessment using the CCK-8 assay in pulmonary fibroblasts isolated from BLM-treated mice after FasL treatment. (C) Caspase 3 activity analysis in TGF-β1-treated pulmonary fibroblasts, along with or without FasL treatment. (D) Cell viability assessment using the CCK-8 assay in TGF-β1-treated pulmonary fibroblasts, along with or without FasL treatment. (E) Western blot analysis of FoxM1 expression in pulmonary fibroblasts transfected with or without LV-FoxM1. (F) Western blot analysis of FoxM1 expression in pulmonary fibroblasts transfected with or without FoxM1 siRNA (si-FoxM1). (G, H) Caspase 3 activity and cell viability assessment in pulmonary fibroblasts transfected with or without LV-FoxM1, following with or without FasL treatment. (I, J) Caspase 3 activity and cell viability assessment in pulmonary fibroblasts transfected with or without si-FoxM1, following with or without FasL treatment. All data (n = 3, ∗ P < 0.05) were presented as the means ± SEM. Paired t -test was used for statistical analysis.

Article Snippet: For the induction of pulmonary fibrosis, a single dose of bleomycin (BLM, Nippon Kayaku, Tokyo, Japan) at 5 mg/kg, dissolved in 50 μl of sterile saline, was administered intratracheally to the treatment group.

Techniques: Expressing, Activity Assay, Isolation, CCK-8 Assay, Western Blot, Transfection

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Impairing nuclear translocation of FoxM1 suppresses fibroblast activation and protects mice from bleomycin-induced pulmonary fibrosis. (A) Western blot analysis was performed to assess the nuclear expression levels of FoxM1 in pulmonary fibroblasts isolated from BLM-treated mice. n = 3, ∗ P < 0.05. (B) Western blot analysis of nuclear FoxM1, CTHRC1, α-SMA, and Collagen I expression in TGF-β1-treated pulmonary fibroblasts accompany with or without RCM-1 treatment. n = 3, ∗ P < 0.05. (C, D) EdU assay for the proliferation of TGF-β1-treated pulmonary fibroblasts accompany with or without RCM-1 treatment. n = 3, ∗ P < 0.05. (E) Hematoxylin–eosin (H&E) and masson's trichrome staining for the lung tissues from BLM-treated mice injected with or without RCM-1. (F, G) The ashcroft score (n = 6, ∗ P < 0.05.) and hydroxyproline contents (n = 6, ∗ P < 0.05.) in the lung tissues of BLM-treated mice injected with or without RCM-1. (H) The survival of BLM-treated mice injected with or without RCM-1. n = 18. (I) Western blot analysis of FoxM1, CTHRC1, α-SMA, and Collagen I expression in the lung tissues from BLM-treated mice injected with or without RCM-1. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (A) and one-way ANOVA with Tukey's post-hoc test (B, D, F–I) were used for statistical analysis.

Article Snippet: For the induction of pulmonary fibrosis, a single dose of bleomycin (BLM, Nippon Kayaku, Tokyo, Japan) at 5 mg/kg, dissolved in 50 μl of sterile saline, was administered intratracheally to the treatment group.

Techniques: Translocation Assay, Activation Assay, Western Blot, Expressing, Isolation, EdU Assay, Staining, Injection

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Acetylation of FoxM1 is required for the activation of pulmonary fibroblasts. (A, B) Western blot analysis of FoxM1 expression in the cytoplasm and nucleus of CHX-treated pulmonary fibroblasts along with or without MG132 treatment at indicated time. n = 3, ∗ P < 0.05. (C) Western blot analysis of FoxM1 expression in CHX-treated pulmonary fibroblasts along with or without TGF-β1 treatment. n = 3, ∗ P < 0.05. (D) Western blot analysis of the acetylation levels of FoxM1 in pulmonary fibroblasts isolated from bleomycin (BLM)-treated mice. n = 3, ∗ P < 0.05. (E) Western blot analysis of the acetylation levels of FoxM1 in pulmonary fibroblasts treated with or without TGF-β1. n = 3, ∗ P < 0.05. (F, G) Western blot analysis of FoxM1 acetylation, CTHRC1, α-SMA, and Collagen I expression in pulmonary fibroblasts treated with TSA (50 nM), or NAM (1 mM) for 24 h, or not. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (D, E) and one-way ANOVA with Tukey's post-hoc test (B, C, G) were used for statistical analysis.

Article Snippet: For the induction of pulmonary fibrosis, a single dose of bleomycin (BLM, Nippon Kayaku, Tokyo, Japan) at 5 mg/kg, dissolved in 50 μl of sterile saline, was administered intratracheally to the treatment group.

Techniques: Activation Assay, Western Blot, Expressing, Isolation

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Sirt3-dependent deacetylation of FoxM1 regulates the stability of FoxM1. (A) The Pearson's correlation analysis of COL1A1 expression with SIRTs expression based on the RNA-seq results of GSE2052 from GEO database. (B) Western blot analysis of SIRT3 expression in the lung tissues from bleomycin (BLM)-treated mice. n = 3, ∗ P < 0.05. (C) Western blot analysis of the acetylation levels of FoxM1 in SIRT3 flox/flox mice intratracheally injected with AAV-Cre. n = 3, ∗ P < 0.05. (D) Western blot analysis was performed to assess the acetylation status of FoxM1 in pulmonary fibroblasts following transfection with Sirt3 siRNA (si-Sirt3). n = 3, ∗ P < 0.05. (E) Western blot analysis of FoxM1 expression in CHX-treated pulmonary fibroblasts transfected with or without si-Sirt3. n = 3, ∗ P < 0.05. (F) Western blot analysis of CTHRC1, α-SMA and Collagen I expression in pulmonary fibroblasts transfected with or without si-Sirt3. n = 3, ∗ P < 0.05. (G) EdU assay for the proliferation of pulmonary fibroblasts transfected with or without si-Sirt3. n = 3, ∗ P < 0.05. (H) Western blot analysis of FoxM1 acetylation, CTHRC1, α-SMA, and Collagen I expression in TGF-β1-treated pulmonary fibroblasts transfected with or without LV-Sirt3. n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. Paired t -test (B-D, F, G) and one-way ANOVA with Tukey's post-hoc test (E, H) were used for statistical analysis.

Article Snippet: For the induction of pulmonary fibrosis, a single dose of bleomycin (BLM, Nippon Kayaku, Tokyo, Japan) at 5 mg/kg, dissolved in 50 μl of sterile saline, was administered intratracheally to the treatment group.

Techniques: Expressing, RNA Sequencing, Western Blot, Injection, Transfection, EdU Assay

Journal: Redox Biology

Article Title: SIRT3-mediated deacetylation of FoxM1 prevents pulmonary fibrosis via modulating the activation of pulmonary fibroblasts

doi: 10.1016/j.redox.2026.104108

Figure Lengend Snippet: Nicotinamide riboside protects mice from bleomycin-induced pulmonary fibrosis via activation of SIRT3. (A) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in TGF-β1-treated pulmonary fibroblasts accompany with or without NR treatment. n = 3, ∗ P < 0.05. (B) Cell viability assessment using the CCK-8 assay in pulmonary fibroblasts treated as in A n = 6, ∗ P < 0.05. (C) The survival of BLM-treated mice oral gavaged with or without NR. n = 18. (D) Hematoxylin–eosin (H&E) and masson's trichrome staining for the lung tissues from mice treated as in C. (E) The ashcroft score of mice treated as in C n = 6, ∗ P < 0.05. (F) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in the lung tissues from mice treated as in C n = 3, ∗ P < 0.05. (G) Western blot analysis of CTHRC1, SIRT3, FoxM1, α-SMA and Collagen I expression in pulmonary fibroblasts isolated from the lung tissues of mice treated as in C n = 3, ∗ P < 0.05. All data were presented as the means ± SEM. One-way ANOVA with Tukey's post-hoc test was used for statistical analysis.

Article Snippet: For the induction of pulmonary fibrosis, a single dose of bleomycin (BLM, Nippon Kayaku, Tokyo, Japan) at 5 mg/kg, dissolved in 50 μl of sterile saline, was administered intratracheally to the treatment group.

Techniques: Activation Assay, Western Blot, Expressing, CCK-8 Assay, Staining, Isolation

Journal: Journal of Translational Medicine

Article Title: Vitamin D attenuates the progression of pulmonary fibrosis via inhibiting thymidine kinase 1/PFKFB3-driven glycolysis

doi: 10.1186/s12967-025-07298-1

Figure Lengend Snippet: Combined Vitamin D and 3PO alleviates bleomycin-induced pulmonary fibrosis in vivo. ( A ) Strategy for administering Vitamin D, 3PO, or a synergistic combination in a bleomycin-induced pulmonary fibrosis mouse model. ( B ) Body weight (g), mean ± SEM. ( C ) H&E staining, Masson staining, and immunohistochemical staining of TK1. ( D ) Hydroxyproline content was quantified in lung tissues ( n = 4), with * P < 0.05, ** P < 0.01, and *** P < 0.001 indicating significance. ( E - F ) Lactate concentrations were measured in the alveolar lavage fluid and lung tissue of mice in the indicated groups ( n = 4), with ** P < 0.01 and *** P < 0.001. ( G ) Expression levels of Fibronectin, Collagen I, α-SMA, TK1, and PFKFB3 were assessed by western blot analysis. GAPDH served as a loading control. The experiments were repeated three times with consistent results. ( H - I ) Glucose consumption and lactate levels were measured in primary fibroblast cultures treated with Vitamin D or 3PO ( n = 4), with * P < 0.05 and ** P < 0.01. For B , a 2-way repeated measures ANOVA was used. For D , E , F , H , and I , one-way ANOVA with the Bonferroni multiple comparisons test was applied. Data are expressed as mean ± SD. Source data are provided in the Source data file.

Article Snippet: Mice in the bleomycin-treated cohorts received a single intratracheal aerosol administration of bleomycin (2.5 mg/kg; MedChemExpress, USA).

Techniques: In Vivo, Staining, Immunohistochemical staining, Expressing, Western Blot, Control