Gamma-aminobutyric acid enhances miR-21-5p loading into adipose-derived stem cell extracellular vesicles to alleviate myocardial ischemia-reperfusion injury via TXNIP regulation

Figure 6 Extracellular vesicles derived from adipose-derived mesenchymal stem cells from inguinal adipose tissue-associated microRNA-21-5p alleviates hypoxia and reoxygenation-mediated myocardial injury and mitochondrial dysfunctions by downregulating thioredoxin-interacting protein. A: Experiment design. Adipose-derived mesenchymal stem cells (ADSCs) were isolated from both microRNA-21 (miR-21) knockout (KO) and miR-21^f/f mouse inguinal adipose tissue (IAT), and extracellular vesicles (EVs) were extracted respectively. Neonatal mouse cardiomyocytes (NMCMs) and NMCMs^{TXNIP KO} were respectively treated with gamma-aminobutyric acid (GABA)-induced ADSCs from IAT (GABA-EVs^IAT) and GABA-EVs^{IAT miR 21KO}; B: Representative micrographs of NMCMs stained with thioredoxin-interacting protein (TXNIP) antibody (n = 5; scale bars = 200 μm); C: The percentage of AnnexinV+ NMCMs was calculated using flow cytometry (n = 5); D: Representative micrographs of isolated NMCMs stained with JC1 probe (n = 5; scale bars = 200 μm); E: Real-time oxygen consumption rates (OCRs) and calculated basal and maximal respiration rates in NMCMs (n = 3); F: Representative micrographs of isolated NMCMs stained with mitosox probe (n = 5; scale bars = 200 μm). ^bP < 0.01, ^cP < 0.001, ^dP < 0.0001. H/R: Hypoxia and reoxygenation; TUNEL: Terminal deoxynucleotidyl transferase dUTP nick end-labeling.