MiR-539-5p alleviates sepsis-induced acute lung injury by targeting ROCK1

Abstract

Introduction. Sepsis-induced acute lung injury (ALI) is an inflammatory process involved with simultaneous production of inflammatory cytokines and chemokines. In this study, we investigated the regulatory role of miR-539-5p in sepsis-induced ALI using a mouse model of cecal ligation puncture (CLP) and an in vitro model of primary murine pulmonary microvascular endothelial cells (MPVECs). Material and methods. Adult male C57BL/6 mice were intravenously injected with or without miR-539-5p agomir or scrambled control one week before CLP operation. MPVECs were transfected with miR-539-5p mimics or control mimics, followed by lipopolysaccharide (LPS) stimulation. ROCK1 was predicted and confirmed as a direct target of miR-539-5p using dual-luciferase reporter assay. In rescue experiment, MPVECs were co-transfected with lentiviral vector expressing ROCK1 (or empty vector) and miR-539-5p mimics 24 h before LPS treatment. The transcriptional activity of caspase-3, the apoptosis ratio, the levels of miR-539-5p, interleukin-1b (IL-1b), interleukin-6 (IL-6), and ROCK1 were assessed. Results. Compared to sham group, mice following CLP showed pulmonary morphological abnormalities, elevated production of IL-1b and IL-6, and increased caspase-3 activity and apoptosis ratio in the lung. In MPVECs, LPS stimulation resulted in a significant induction of inflammatory cytokine levels and apoptosis compared to untreated cells. The overexpression of miR-539-5p in septic mice alleviated sepsis-induced pulmonary injury, apoptosis, and inflammation. MiR-539-5p also demonstrated anti-apoptotic and anti-inflammatory effect in LPS-treated MPVECs. The upregulation of ROCK1 in MPVECs recovered miR-539-5p-suppressed caspase-3 activity and proinflammatory cytokine production. Conclusion. In conclusion, miR-539-5p alleviated sepsis-induced ALI via suppressing its downstream target ROCK1, suggesting a therapeutic potential of miR-539-5p for the management of sepsis-induced ALI.

miR‑26a‑5p alleviates lipopolysaccharide‑induced acute lung injury by targeting the connective tissue growth factor

“…A recent study has indicated that miR-802 was expressed at a low level in LPS-induced ALI and alleviated LPS-induced ALI by targeting Peli2 (20). Another recent study has revealed that miR-539-5p was underexpressed in sepsis-induced ALI and could protect mice from ALI (21). In the present study, the expression of miR-26a-5p was downregulated in LPS-induced ALI models in vivo and in vitro.…”

“…Previous studies have indicated that ALI is featured with the production of inflammatory factors, inflammatory cell infiltration and pulmonary epithelial cell apoptosis (24,25). Evidence from several clinical studies has indicated that TNF-α, IL-1β and IL-6 are the main inflammatory cytokines actively secreted in response to the inflammatory cascade in ALI (21,26). Previous studies have indicated that the excess inflammatory cytokines, such as TNF-α, IL-6 and IL-1β, ultimately cause cell death via apoptosis and pyroptosis (27).…”

CircC3P1 attenuated pro‐inflammatory cytokine production and cell apoptosis in acute lung injury induced by sepsis through modulating miR‐21

“…Proteins extracted from MPVEC lysates were analysed by 8% SDS‐PAGE gel and transferred to PVDF membranes as previously described 22 . Then, membranes were incubated with primary antibodies (1:1000, Abcam) for 6 hours at 4°C, including p‐AKT, AKT and β‐actin.…”

MicroRNA-490-3p inhibits inflammatory responses in LPS-induced acute lung injury of neonatal rats by suppressing the IRAK1/TRAF6 pathway

“…Similarly, multiple miRNAs have been revealed to regulate inflammation in ALI. Meng et al (29) revealed that miR-539-5p relieved sepsis-induced ALI by targeting the protein Rho associated coiled-coil containing protein kinase 1. Yang et al (30) revealed that miR-142a-3p alleviated ALI induced by E. coli lipopolysaccharides by acting on TGF-β-activated kinase 1 and MAP3K7-binding protein 2.…”