Figures & data
Figure 1. Sema3A is downregulated in the lungs of rats with ARDS. A, Sema3A (NM_017310) as the most significantly differentially expressed gene between ARDS and control lungs in the GSE57011 dataset; B, morphological changes in rat lung tissues observed by HE staining; C, Sema3A expression in rat lung tissues examined by IHC; D, single cell sequencing analysis of Sema3A localisation in the lung predicted in The Human Protein Atlas. In each group, n = 6. Differences were analysed by the unpaired t test (C). * p < 0.05.
Figure 2. Sema3A elevation alleviates inflammation and oxidative stress in rat PMVECs. A, protein level of Sema3A in PMVECs after oe-Sema3A administration determined by WB analysis; B, viability of PMVECs examined by CCK-8 assay; C, apoptosis of PMVECs examined by flow cytometry; D, angiogenesis of PMVECs analysed by tube formation assay; E, concentrations of inflammatory cytokines IL-1β, IL-6 and TNF-α in the culture supernatant of PMVECs detected by ELISA kits; F, protein levels of VE-cadherin and α-E-catenin in PMVECs determined by WB analysis; G, concentrations of oxidative stress-related markers MDA, MPO and SOD in PMVECs detected by ELISA kits. Three biological replicates were performed. Differences were analysed by the one-way ANOVA (A-G). * p < 0.05.
Figure 3. Sema3A restoration alleviates inflammation and oxidative stress in rat lung tissues. A, lung injury in rats determined by HE staining; B, lung permeability analysed by Evans blue staining; C, apoptosis of lung endothelial cells examined by TUNEL assay; D, pulmonary edoema evaluated by the W/D weight ratio; E, protein concentration in rat BALF examined by bicinchoninic acid and the number of neutrophils in rat BALF examined by Wright’s staining; F, concentrations of IL-1β, IL-6, TNF-α and VEGF in rat BALF determined by ELISA kits; G, concentrations of TF and TAT in rat BALF determined by ELISA kits; H, levels of MDA, MPO, and SOD rat BALF determined by ELISA kits. In each group, n = 6. Differences were analysed by the one-way ANOVA (B-H). * p < 0.05.
Figure 4. Sema3A suppresses the ERK/JNK signalling pathways. A, phosphorylation of ERK and JNK in PMVECs detected by WB analysis; B, phosphorylation of ERK and JNK in rat lung tissues detected by WB analysis. For cellular experiments, three biological replicates were performed. For animal experiments, n = 6 in each group. Differences were analysed by the one-way ANOVA (A-B). * p < 0.05.
Figure 5. Activation of ERK/JNK increases inflammation and oxidative stress in PMVECs. A, phosphorylation of ERK and JNK in PMVECs after LM22B-10 or Juglanin treatment; B, viability of PMVECs examined by CCK-8 assay; C, apoptosis of PMVECs examined by flow cytometry; D, concentrations of IL-1β, IL-6 and TNF-α in the culture supernatant of PMVECs detected by ELISA kits; E, concentrations of oxidative stress-related markers MDA, MPO and SOD in PMVECs detected by ELISA kits. Three biological replicates were performed. Differences were analysed by the one-way ANOVA (A-E). * p < 0.05.
Data availability statement
All data generated or analysed during this study are included in this published article.