Fig. 8
From: Autophagy regulates the effects of ADSC-derived small extracellular vesicles on acute lung injury
The effect of ADSC-sEVs with or without autophagy inhibition on LPS-induced acute lung injury. A Lung histology visualized with hematoxylin and eosin staining. B Microscopic injury of the lung was statistically scored. Administration of ADSC-sEV effectively attenuated LPS-triggered lung injury. The effect of ADSCsiRNA−NC-sEV on lung injury was similar with that of ADSC-sEV. However, the protective effect of ADSCsiATG5-sEV was much worse than that of ADSC-sEV or ADSCsiRNA−NC-sEV. The results are presented as the mean ± SD (the lung injury score, control, 0.33 ± 0.52 vs. LPS, 3.33 ± 0.52 vs. LPS + ADSC-sEV, 1.17 ± 0.41 vs. LPS + ADSCsiRNA−NC-sEV, 1.17 ± 0.41 vs. ADSCsiATG5-sEV, 2.50 ± 0.55). C Lung water content was tested by detecting the wet/dry weight ratio. LPS challenge increased the lung wet/dry weight ratio, which was effectively attenuated by ADSC-sEV treatment. However, the inhibitory action of ADSCsiATG5-sEV on LPS-induced lung edema was much weaker than that of ADSC-sEVs (the lung wet/dry weight ratio, control, 4.05 ± 0.73 vs. LPS, 7.39 ± 0.29 vs. LPS + ADSC-sEV, 5.27 ± 0.23 vs. LPS + ADSCsiRNA−NC-sEV, 5.34 ± 0.15. vs. ADSCsiATG5-sEV, 6.63 ± 0.54). D Lung microvascular permeability was assessed by detecting the changes in total protein concentration in bronchoalveolar lavage fluid (BALF). LPS markedly increased protein leakage in BALF, which was significantly attenuated by ADSC-sEV or ADSCsiATG5−NC-sEV treatment. However, the inhibitory effect of ADSCsiATG5-sEV on protein leakage in BALF was markedly weaker than that of ADSC-sEV (the total protein concentration in BALF (μg/ml), control, 11.25 ± 0.33 vs. LPS, 36.71 ± 0.94 vs. LPS + ADSC-sEV, 23.34 ± 0.14 vs. LPS + ADSCsiRNA−NC-sEV, 23.32 ± 0.62. vs. ADSCsiATG5-sEV, 29.62 ± 0.44). E, F ELISA assay was used to detect the concentrations of TNF-α and IL-1β in BALF from mouse lungs that underwent LPS treatment. LPS significantly increased the production of TNF-α and IL-1β, which was effectively inhibited by ADSC-sEV or ADSCssiATG5-sEV treatment. However, the inhibitory effects of ADSCsiATG5-sEVs on the production of TNF-α and IL-1β were significantly weaker than those of ADSC- sEVs (the concentration of TNF-α (pg/ml), control, 1291.21 ± 34.34 vs. LPS, 3011.75 ± 91.08 vs. LPS + ADSC-sEV, 2016.01 ± 28.66 vs. LPS + ADSCsiRNA−NC-sEV, 2107.39 ± 83.85 vs. ADSCsiATG5-sEV, 2647.53 ± 111.61). The concentration of IL-1β (pg/ml), control, 362.99 ± 22.17 vs. LPS, 624.98 ± 16.92 vs. LPS + ADSC-sEV, 423.24 ± 18.47 vs. LPS + ADSCsiRNA−NC-sEV, 425.23 ± 16.06 vs. ADSCsiATG5-sEV, 536.40 ± 14.23). The results are presented as the mean ± SD. The results are presented as the mean ± SD (n = 18/group, 6 for H&E staining and pathological scores; 6 for wet/dry weight ratio; 6 for BALF collection and assessment of total protein level and TNF-α and IL-1β concentration)