Research Article
Cardiac Fibroblasts Promote Ferroptosis in Atrial Fibrillation by Secreting Exo-miR-23a-3p Targeting SLC7A11
Figure 5
Rapid pacing primary cardiac fibroblast-derived exosomes aggravate the ferroptosis of h9c2 cells. (a) Morphology of primary cardiac fibroblast-derived exosomes (CF-exos) using transmission electron microscopy. (b) Mean exosomes diameter shown by ZetaView System. (c) Exosomes were isolated from culture supernatant from CFs, dyed with phalloidin (green) and cocultured with h9c2 cells, then dyed with PKH26 (red) and viewed with fluoroscopy. (d, e) Representative gel bands depicting FTH1, GPX4, and SLC7A11 proteins expression in h9c2 cells with or without Fer-1 incubated with CF-exos for 48 h. CFs treated with or without GW4869 and stimulated by rapid pacing for 48 h and its exosomes were isolated from culture supernatant. (f) Total iron level in h9c2 cells incubated with CF-exos. (g) MDA level in h9c2 cells incubated with CF-exos. (h, i) Mitochondrial membrane potential was detected through flow cytometry in h9c2 cells incubated with CF-exos. (j, k) Reactive oxygen species was detected through flow cytometry in h9c2 cells incubated with CF-exos. Data are presented as the mean ± SD, =3. Statistical significance was determined using one-way ANOVA with a post hoc Dunnett test. , , and vs. control group; , , and vs. pacing-CF-exos group. Abbreviations: control-CF-exos: h9c2 cells incubated with control-CF-exos; pacing-CF-exos: h9c2 cells incubated with CF-exos stimulated by rapid pacing 48 h; pacing-CF-exos+Fer-1: h9c2 cells treated with Fer-1 (20 μM) and incubated with CF-exos stimulated by rapid pacing 48 h; GW4869-pacing-CF-exos: h9c2 cells incubated with CF-exos treated with GW4869 (20 μM) and stimulated by rapid pacing 48 h.
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