human pulmonary microvascular endothelial cells pmvecs  (PromoCell)

Journal: Biology Direct

Article Title: Enhanced itaconic acid secretion from macrophages mediates the protection of mesenchymal stem cell-derived exosomes on lipopolysaccharide-induced acute lung injury mice

doi: 10.1186/s13062-024-00586-8

Figure Lengend Snippet: AMs aggravate LPS-induced injury in PMVECs via M1 polarization-mediated inflammatory response. (A) PMVECs were cultured in the presence of AMs or alone, followed by 100 ng/mL LPS treatment. Equivalent PBS was used as the control. After 24 h of culture, (B) PMVEC permeability was assessed using HRP leakage assays. (C) Western blot analysis revealed the relative expressions of both Claudin-5 and ZO-1 proteins, with GAPDH as the internal reference. (D) Annexin V-FITC/PI staining and (E) TUNEL assays demonstrated the apoptosis rate of PMVECs. (E) ELISA was used for inflammatory cytokines (F) IL-6 and (G) TNF-α concentration determination. (H) The western blot analysis for iNOS and Arg-1 represented the level of M1 polarization. Data are presented as mean ± SEM. Statistical significance was determined using appropriate methods, with * p < 0.05, *** p < 0.001. PMVECs: pulmonary microvascular endothelial cells; AMs: alveolar macrophages; HRP: horseradish peroxidase

Article Snippet: Mouse pulmonary microvascular endothelial cells (PMVECs; CP-M001) were purchased from Procell Life Science (Wuhan, China) and maintained in endothelial cell growth medium (Procell) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (p/s).

Techniques: Cell Culture, Control, Permeability, Western Blot, Staining, TUNEL Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Biology Direct

Article Title: Enhanced itaconic acid secretion from macrophages mediates the protection of mesenchymal stem cell-derived exosomes on lipopolysaccharide-induced acute lung injury mice

doi: 10.1186/s13062-024-00586-8

Figure Lengend Snippet: MSC-exos attenuate LPS-induced PMVEC injury by promoting M2 polarization of AMs. MSC-exos were added to the LPS-treated co-culture system to evaluate their effects on PMVECs. (A) PMVEC permeability was assessed using HRP leakage assays. (B) Western blot analysis was performed to detect the expression levels of tight junction proteins Claudin-5 and ZO-1. (C) Annexin V-FITC/PI staining and (D) TUNEL assays demonstrated the apoptosis rate of PMVECs to determine the anti-apoptosis effect of MSC-exos. (E-F) ELISA for inflammatory cytokines IL-6 and TNF-α in cell supernatant. (G) Western blot analysis for M1 polarization markers assessed the relative levels of iNOS and Arg-1 proteins. (H) M2 polarization was determined via flow cytometry by calculate the percentage of CD206-positive cells. (I) Gas chromatography-mass spectrometry was used to measure ITA concentration. Data are presented as mean ± SEM. Statistical significance was determined using appropriate methods, with * p < 0.05, ** p < 0.01, and *** p < 0.001. MSC-exos: mesenchymal stem cell-derived exosome; PMVECs: pulmonary microvascular endothelial cells; AMs: alveolar macrophages; HRP: horseradish peroxidase

Article Snippet: Mouse pulmonary microvascular endothelial cells (PMVECs; CP-M001) were purchased from Procell Life Science (Wuhan, China) and maintained in endothelial cell growth medium (Procell) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (p/s).

Techniques: Co-Culture Assay, Permeability, Western Blot, Expressing, Staining, TUNEL Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Gas Chromatography, Mass Spectrometry, Concentration Assay, Derivative Assay