Research Paper Volume 13, Issue 4 pp 6156—6170

Extracellular vesicles from anoxia preconditioned mesenchymal stem cells alleviate myocardial ischemia/reperfusion injury

Analysis of pyroptosis and apoptosis markers in EV-treated, AR-exposed cardiomyocytes. (A, B) Western blot analysis of TXNIP and cleaved-caspase-1 in AR-exposed cardiomyocytes incubated with EVs derived from normoxic (NC-EVs) or anoxic (Int-EVs) ADSCs. The NC-EVs group was compared with the control AR group and the Int-EVs group was compared with the NC-EVs group (*P C, D) Western blot analysis of BAX, cleaved-caspase-3, GATA4, and Bcl-2 in cardiomyocytes treated with AR. Comparisons were made between NC-EVs and AR and between Int-EVs and NC-EVs groups (*P E, F) Apoptosis detection in AR-exposed cardiomyocytes (Annexin V/PI assay). The NC-EVs group was compared with the AR group and the Int-EVs group was compared with the NC-EVs group (*P G, H) Relative expression of cleaved GSDMD and total GSDMD in cardiomyocytes and RAW264.7 cells treated with or without AR (*P

Figure 2. Analysis of pyroptosis and apoptosis markers in EV-treated, AR-exposed cardiomyocytes. (A, B) Western blot analysis of TXNIP and cleaved-caspase-1 in AR-exposed cardiomyocytes incubated with EVs derived from normoxic (NC-EVs) or anoxic (Int-EVs) ADSCs. The NC-EVs group was compared with the control AR group and the Int-EVs group was compared with the NC-EVs group (*P < 0.05, **P < 0.01, *** P < 0.001, **** P < 0.0001, n = 4). (C, D) Western blot analysis of BAX, cleaved-caspase-3, GATA4, and Bcl-2 in cardiomyocytes treated with AR. Comparisons were made between NC-EVs and AR and between Int-EVs and NC-EVs groups (*P < 0.05, **P < 0.01, *** P < 0.001, **** P < 0.0001, n = 4). (E, F) Apoptosis detection in AR-exposed cardiomyocytes (Annexin V/PI assay). The NC-EVs group was compared with the AR group and the Int-EVs group was compared with the NC-EVs group (*P < 0.05, **P < 0.01, *** P < 0.001, **** P < 0.0001, n = 4). (G, H) Relative expression of cleaved GSDMD and total GSDMD in cardiomyocytes and RAW264.7 cells treated with or without AR (*P < 0.05, **P < 0.01, *** P < 0.001, **** P < 0.0001, compared with the control CM group; n = 4); (#P < 0.05, ##P < 0.01,###P < 0.001, #### P < 0.0001, compared with the CM + AR group; n = 4); (&P < 0.05, &&P < 0.01, &&& P < 0.001, &&&&P < 0.0001, compared with the RAW264.7 group; n = 4).