Small Extracellular Vesicles Secreted by iPSC-Derived MSCs Ameliorate Pulmonary Inflammation and Lung Injury Induced by Sepsis through Delivery of miR-125b-5p

Figure 6

miR-125b-5p transmitted by iMSC-sEV suppresses NF-κB signaling in alveolar macrophages by directly targeting TRAF6. (a) Online TargetScan v7.2 predicted miR-125b-5p targeted to TRAF6. (b) Protein expression of TRAF6 in LPS-treated NR8383 cells under iMSC-sEV coculture. (c, d) The luciferase reporter assay examined the luciferase activity of the indicated reporter vectors in 293 T cells cotransfected with miR-125b-5p mimics or NC mimics. (e) Western blot analysis detected the protein levels of TRAF6 after transfection with the miR-125b-5p mimic or miR-125b-5p inhibitor. β-Actin was used as an internal control. (f) ELISA examined the level of inflammatory cytokine expression in LPS-treated NR8383 cells when overexpressed with TRAF6. (g) The western blot analyzed the protein level of p-p65 in LPS-treated NR8383 cells when overexpressed with TRAF6. Rescue of TRAF6 expression dramatically inhibited iMSC-sEV to inactivate p65 phosphorylation. β-Actin was used as an internal control. (h) The concept map demonstrating the role and functional mechanism of iMSC-sEV on alleviating septic lung injury. Data presented as mean ± SD. compared within two groups. NS, no significant difference, n = 3 per group.

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