fig5

Figure 5. NRP1 affects apoptosis, migration proteins, and vascular permeability through the p130Cas/MMPs pathway. (A) Transfection with NRP1-overexpressing plasmid rescued LPS-induced imbalance in the Bcl-2/Bax ratio. Quantitative analysis of the Bcl-2/Bax ratio; (B) Reduced Cyt c expression following upregulation of the Bcl-2/Bax ratio. Quantitative analysis of Cyt c band intensity; (C) Downregulated levels of Cleaved Caspase-3/9. Quantitative analysis of Cleaved Caspase-3 band intensity. Quantitative analysis of Cleaved Caspase-9 band intensity. All experiments were independently repeated three times; (D) NRP1-overexpressing MSC-Exo significantly promoted MMP1 and MMP9 expression; (E) Tight junction-associated proteins OCLN and ZO-1 were synchronously upregulated; (F) Immunofluorescence confirmed co-localization of NRP1 with PDGFR-α; (G) Western blot validated NRP1-mediated activation of the p130Cas/MMPs signaling axis in combination with exogenous PDGF-BB (30 ng/mL). All experiments were independently repeated three times. Data were presented as mean ± SD. Statistical analysis was performed using one-way ANOVA followed by Tukey’s multiple comparisons test. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001. NRP1: Neuropilin-1; MSC-Exo: mesenchymal stem cell-derived exosomes; Con-Exo: exosomes derived from control mesenchymal stem cells; OE-Exo: exosomes derived from NRP1-overexpressing mesenchymal stem cells; Si-Exo: exosomes derived from NRP1-silenced mesenchymal stem cells; LPS: lipopolysaccharide; NC: normal control; Bcl-2: B-cell lymphoma 2; Bax: Bcl-2-associated X protein; Cyt c: cytochrome c; MMP-1: matrix metalloproteinase-1; MMP-9: matrix metalloproteinase-9; OCLN: occludin; ZO-1: zonula occludens-1; PDGFR-α: platelet-derived growth factor receptor alpha; PDGF-BB: platelet-derived growth factor-BB; p130Cas: Crk-associated substrate (130 kDa); p-p130Cas: phosphorylated Crk-associated substrate; DAPI: 4’,6-diamidino-2-phenylindole; SD: standard deviation; ANOVA: analysis of variance.