Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: Dysregulation of BMP9/ALK1 signaling and inflammation in refractory ulcerative colitis (rUC). ( A ) Heatmap depicting serum expression levels of BMP family members in healthy controls, non-rUC, and rUC patients ( n = 3 per group). Data were Z-score normalized and hierarchically clustered (red, high expression; blue, low expression). ( B ) Baseline serum levels of BMP9 and BMP10 in healthy controls ( n = 50), non-rUC patients ( n = 47), and rUC patients ( n = 48). ( C ) Spearman correlation analyses between baseline serum BMP9 levels and clinical disease activity indices, including baseline Modified Mayo Score, baseline UCEIS, post-treatment Modified Mayo Score, and post-treatment UCEIS, in patients with UC ( n = 95). ( D ) Colonic mucosal mRNA expression levels of ALK1, IL-6, TNF-α, and CCL2 in healthy controls, non-rUC, and rUC patients ( n = 4 per group). Statistical annotations for ( B , D ): (Normalized to GAPDH; Mean ± SD; Statistical significance determined by one-way ANOVA with Tukey’s post hoc test: ** p < 0.01, *** p < 0.001, ns: not significant).

Article Snippet: Immediately after seeding, HIMECs were treated with the following stimuli: recombinant human BMP9 (rhBMP9; 0–10 ng/mL), TNF-α (20 ng/mL; Novoprotein, Suzhou, China, #C008), and ML347 (150 nM; MCE, Monmouth Junction, NJ, USA, #HY-12274).

Techniques: Expressing, Activity Assay, Modification

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9 attenuates DSS-induced colitis in mice. ( A ) Schematic of experimental design: Acute colitis was induced in C57BL/6 mice by 3% DSS in drinking water for 7 days. The BMP9 treatment group received intraperitoneal injections of recombinant murine BMP9 (200 ng/day), while the DSS group and the control group received PBS ( n = 14 per group). ( B ) Serum BMP9(ng/mL) concentrations measured by ELISA. ( C ) Representative images of colons and quantitative analysis of colon length (cm). ( D ) Colonoscopy images (upper), H&E-stained colon sections (lower; scale bars = 100 μm), and histopathological scores. ( E ) (Upper) Dynamic body weight changes and (Lower) Disease Activity Index (DAI) scores. Data expressed as mean ± SD; * p < 0.05, two-way repeated measures ANOVA. ( F ) (Left) Relative Alk1 mRNA expression in colon tissues (RT-qPCR normalized to Gapdh). (Right) ALK1 protein concentrations (quantified by ELISA). Data presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 ns: not significant). ( G ) Relative mRNA expression levels of IL-1β, Ccl2, Col1a1, and Col3a1 (RT-qPCR; mean ± SD). ( H ) Western blot analysis of CCL2, TGF-β, and α-SMA protein expression in colon tissues and quantitative analysis of band intensities. ( I ) Western blot detection of p-Smad1, total Smad1, and VE-cadherin proteins and quantitative analysis of band intensities.

Article Snippet: Immediately after seeding, HIMECs were treated with the following stimuli: recombinant human BMP9 (rhBMP9; 0–10 ng/mL), TNF-α (20 ng/mL; Novoprotein, Suzhou, China, #C008), and ML347 (150 nM; MCE, Monmouth Junction, NJ, USA, #HY-12274).

Techniques: Recombinant, Control, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9 restores intestinal vascular barrier integrity in DSS-induced colitis. ( A ) Representative immunofluorescence images of VE-cadherin (red) and CD31 (green) co-localization in colon tissues (nuclei counterstained with DAPI). ( B ) Left: Schematic of FITC-dextran (4 kDa) permeability assay. Right: Quantified serum FITC fluorescence intensity 60 min post-gavage ( n = 5 per group). ( C ) Left: Schematic of Evans Blue vascular leakage assay. Right: Colonic Evans Blue extravasation quantified by absorbance at 620 nm (mean ± SD; n = 5 per group; ** p < 0.01, *** p < 0.001, ns: not significant; one-way ANOVA with Tukey’s test). ( D ) KEGG pathway analysis of RNA-seq data from colon tissues (DSS + BMP9 groups vs. DSS). ( E ) Volcano plot of differentially expressed genes. Genes highlighted in bold are key IBD-associated downregulated factors.

Article Snippet: Immediately after seeding, HIMECs were treated with the following stimuli: recombinant human BMP9 (rhBMP9; 0–10 ng/mL), TNF-α (20 ng/mL; Novoprotein, Suzhou, China, #C008), and ML347 (150 nM; MCE, Monmouth Junction, NJ, USA, #HY-12274).

Techniques: Immunofluorescence, Permeability, Fluorescence, RNA Sequencing

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9/ALK1 signaling modulates neutrophil migration and endothelial tube formation. ( A ) Schematic representation of the neutrophil migration assay. ( B ) (Left) Representative fluorescence micrographs demonstrating Calcein-AM-labeled neutrophil migration through 3 μm pore Transwell inserts toward HIMECs pretreated for 2 h with: vehicle control (0 ng/mL BMP9), BMP9 (0.1, 1, or 10 ng/mL), TNF-α (20 ng/mL as positive control), or ALK1 inhibitor ML347 (150 nM). (Right) Quantitative analysis of neutrophil migration rates (mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, ns: not significant by two-way ANOVA with post hoc testing). ( C ) Schematic illustration of the endothelial tube formation assay protocol. ( D ) Representative phase-contrast images of tubular network formation by HIMECs cultured on growth factor-reduced Matrigel under various treatment conditions: vehicle control (0 ng/mL BMP9), BMP9 (0.1, 1, or 10 ng/mL), TNF-α (20 ng/mL), or ML347 (150 nM). ( E ) Quantitative assessment of angiogenic parameters including branch points and nodal junctions (mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA with appropriate post hoc comparisons).

Article Snippet: Immediately after seeding, HIMECs were treated with the following stimuli: recombinant human BMP9 (rhBMP9; 0–10 ng/mL), TNF-α (20 ng/mL; Novoprotein, Suzhou, China, #C008), and ML347 (150 nM; MCE, Monmouth Junction, NJ, USA, #HY-12274).

Techniques: Migration, Fluorescence, Labeling, Control, Positive Control, Endothelial Tube Formation Assay, Cell Culture

Journal: GeroScience

Article Title: Tilorone attenuates high-fat diet-induced hepatic steatosis by enhancing BMP9-Smad1/5/8 signaling.

doi: 10.1007/s11357-025-01685-8

Figure Lengend Snippet: Fig. 8 Summary of the results. After the 10 weeks tilorone treatment, the HFD-induced increases in complex II-linked oxidative phosphorylation and com- plex IV activity reduced and the HFD-induced decreases in BMP9, pSmad1/5/8, and PPARγ levels were normal- ized. PET/MRI showed increased tissue uptake of 18FDG. As a consequence, lipid content of the liver decreased, the glycogen content increased, and blood glucose level, body mass, and liver and adipose tissue weights were normal- ized. HFD, high-fat diet; BMP, bone morphogenetic protein; PPARγ, peroxi- some proliferator-activated receptor gamma; 18FDG, 18F-fluoro-2-deoxyglucose

Article Snippet: Then, the membranes were blocked in Tris buffered saline containing 5% skimmed milk and 0.1% Tween-20 (Sigma-Aldrich) for 1 h at room temperature and were incubated at 4 °C overnight with the following rabbit polyclonal primary antibodies: phospho-Smad1/5/8 (Smad1 [Ser463/465]/Smad5 [Ser463/465]/Smad9 [Ser465/467]; #AB-3848-J; Sigma-Aldrich); Smad1/5/8 (#56,656; Novus Biologicals; Centennial, CO, USA); PPARγ (#2435; Cell Signaling Technology; Danvers; MA; USA); PGC-1α (#2178; Cell Signaling Technology); TOM20 (#42,406; Cell Signaling Technology); BMP6 (#55,421–1-AP; Proteintech; Rosemont; IL, USA); BMP9 (#17,769–1-AP; Proteintech); and GAPDH (#2188; Cell Signaling Technology).

Techniques: Phospho-proteomics, Activity Assay