Circular RNA mmu_circ_0001295 from hypoxia pretreated adipose-derived mesenchymal stem cells (ADSCs) exosomes improves outcomes and inhibits sepsis-induced renal injury in a mouse model of sepsis

Figure 1.. Effects of ADSC exosomes on CLP-induced mortality. (a) The number of particles versus particle size, determined by nanoparticle tracking analysis with ZetaView. (b) Transmission electron micrographs showing ADSC exosome (Exo) or hypoxically pretreated ADSC exosome (HExo) morphology. (c) Western blots of CD63 and CD81 expression in Exo and HExo. (d) Mice were subjected to CLP and treated with HExo or Exo (2 mg protein/kg body weight) or PBS. Survival rates were monitored for a total of 168 h (7 days). **p < 0.01 vs. CLP-PBS. #p < 0.05 vs CLP-Exo. N = 12/group.

Figure 2.. Effects of ADSC exosomes on organ dysfunction and vascular leakage in CLP-induced sepsis. (a-c) Plasma levels of AST (a), ALT (b) and BUN (c), 24 hr after CLP. **p < 0.01, ***p < 0.001 vs sham. ##p < 0.01, ###p < 0.001 vs CLP-PBS. (d) Vascular leakage in the kidney, measured via injection of Evans blue dye at 24 hr after CLP. ***p < 0.001 vs sham. ###p < 0.001 vs CLP-PBS.

Figure 3.. ADSC-exosome treatment decreases inflammatory factor expression in CLP-induced sepsis. (a-d) ELISA detection of the expression of IL-6, IL-1β, TNF-α and MCP-1 in plasma. *p < 0.05, **p < 0.01, ***p < 0.001 vs sham. #p < 0.05, ###p < 0.001 vs CLP-PBS.

Figure 4.. mmu_circ_0001295 plays an important role in HExo’s protective effects against CLP-induced sepsis. (a) Heat map showing the expression of circRNA between Exo and HExo. (b) RT-qPCR detection showing the expression of eight upregulated circRNAs in both Exo and HExo. The data are presented as the mean ± SD. ***p < 0.001 vs Exo. (c) RT-qPCR detection showing the expression of mmu_circ_0001295 in plasma after CLP-induced sepsis. The data are presented as the mean ± SD. ***p < 0.001 vs sham.

Figure 5.. Downregulation of mmu_circ_0001295 decreases the therapeutic effects of ADSC exosomes on organ dysfunction and vascular leakage in CLP-induced sepsis. (a) Mice were subjected to CLP and treated with HExo, si-circ-HExo (2 mg protein/kg body weight) or PBS. Survival rates were monitored for a total of 168 h (7 days). **p < 0.01 vs. CLP-PBS. #p < 0.05 vs CLP-Exo. (b-d) Plasma levels of AST (b), ALT (c) and BUN (d), measured 24 h after CLP. **p < 0.01, ***p < 0.001 vs sham. ###p < 0.001 vs CLP-PBS. (e) Vascular leakage in the kidney, measured via injection of Evans blue dye at 24 hr after CLP. ***p < 0.001 vs sham. ###p < 0.001 vs CLP-PBS.

Figure 6.. Downregulation of mmu_circ_0001295 decreases the inhibitory effects of HExo on inflammatory factor expression in CLP-induced sepsis. (a-d) ELISA detection of the expression of IL-6, IL-1β, TNF-α and MCP-1 in plasma. *p < 0.05, **p < 0.01, ***p < 0.001 vs sham. ###p < 0.001 vs CLP-PBS.