miR-29a-3p in Exosomes from Heme Oxygenase-1 Modified Bone Marrow Mesenchymal Stem Cells Alleviates Steatotic Liver Ischemia-Reperfusion Injury in Rats by Suppressing Ferroptosis via Iron Responsive Element Binding Protein 2

Figure 6

HO-1/BMMSC-derived exosomes inhibit ferroptosis and protect SHP-HR injury by regulating IREB2. (a) HBM-exos were observed as vesicles using transmission electron microscope. (b) Nanoparticle tracking analysis showing that the particle size of the HBM-exos was concentrated in the 100–150 nm range. (c) HBM-exos were positive for CD63, TSG101, and CD9 proteins, but negative for calnexin. (d) Confocal fluorescence microscopy showing that SHPs ingested CM-DiI- (red-) labeled HBM-exos. Evaluation of ferroptosis and cell injury in SHP-HR, with or without HBM-exos. (e) Representative immunofluorescence images of IREB2 showing that HBM-exos downregulated the protein level of IREB2. (f) IREB2/FTH1/TFR1/GPX4 protein levels assessed using western blotting. (g) Ireb2 mRNA level detected using qRT-PCR. (h) The Fe²⁺ levels detected using colorimetry. (i) The MDA content detected using colorimetry. (j) The lipid ROS level detected using C11-BODIPY and flow cytometry. (k) Quantitative analysis of C11-BODIPY fluorescence intensity. (l) Cell viability detected using a CCK-8 kit. (m) Cell mitochondrial membrane potential detected using JC-1 staining and flow cytometry. (n) Quantitative analysis of (m). (o) Cell death ratio detected using PI staining and flow cytometry. (p) Quantitative analysis of (o). HBM-exos, HO-1/BMMSC exosomes. per group. Data are presented as the . ; .

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