Sevoflurane Improves Ventricular Conduction by Exosomes Derived from Rat Cardiac Fibroblasts After Hypothermic Global Ischemia-Reperfusion Injury

Figures

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  Figure 1. Identification of primary cardiac fibroblasts. (A) The morphology and confluency of cardiac fibroblasts at each point (0H, 24H, 48H, 72H are the time after differential adhesion). (B) The purity of cardiac fibroblasts identified by immunofluorescence.

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  Figure 2. Identification and comparison of exosomes. The morphology (A and D), diameters distribution (B and E), and concentration (C and F) of the CFs-Exo and Sev-CFs-Exo were observed by the transmission electron microscope and nanoparticle tracking analysis (The former is CFs-Exo).

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  Figure 3. Effects of CFs-Exo and Sev-CFs-Exo on heart rates. Representative biphasic electrograms from a single channel were obtained from isolated hearts in the control condition (A) and the presence of Sev-CFs-Exo (B) and CFs-Exo (C). The heart rates in the four groups: reperfusion vs balanced perfusion (D), *group IR, #(purple) group N+IR, *(blue) group S+IR. Comparison of heart rates in the four groups at T0 (E), T1 (F), T2 (G), and T3 (H),*vs group IR, #vs group N+IR. Data from n = 8 hearts. **P<0.01, *** or ###P<0.001.

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  Figure 4. Effects of CFs-Exo and Sev-CFs-Exo on reperfusion arrhythmia. (A) Typical electrograms were obtained from spontaneously beating hearts during reperfusion for 30 minutes: normal electrocardiogram (top), ventricular tachycardia (middle) and ventricular fibrillation (bottom). The score basing on the Lambeth conventions (B) and duration (C) of reperfusion arrhythmia. The time for restoration of heartbeats at the point of balanced perfusion (D) and reperfusion (E). *vs group IR, #vs group N+IR. Data from n = 8 hearts. **P<0.01, ***P<0.001.

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  Figure 5. Effects of CFs-Exo and Sev-CFs-Exo on conduction velocity. (A) Classic activation maps were obtained in the four groups at T2. The conduction velocity at T1 (B), T2 (C), and T3 (D), *vs group IR, #vs group N+IR. Reperfusion vs balanced perfusion (E), *group IR, #(purple) group N+IR, *(blue) group S+IR. MAX and MIN are the maximum and minimum values of local activation time respectively, which is contrary to conduction velocity. Data from n = 8 hearts. *** or ###P<0.001.

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  Figure 6. Effects of CFs-Exo and Sev-CFs-Exo on inhomogeneity. (A) Representative inhomogeneity maps were acquired in the four groups at T2. The absolute inhomogeneity and inhomogeneity index respectively at T1 (B and E), T2 (C and F), and T3 (D and G), *vs group IR, #vs group N+IR. Reperfusion vs balanced perfusion (H and I), *group IR, #(purple) group N+IR, *(blue) group S+IR. MAX and MIN are the maximum and minimum values of inhomogeneity respectively. Data from n = 8 hearts. ** or ##P<0.01, *** or ###P<0.001.

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  Figure 7. Effects of CFs-Exo and Sev-CFs-Exo on Cx43 and myocardial ischemia-reperfusion injury. (A and B) The relative expression and location of Cx43 in the four groups assessed by Western blotting assay and immunofluorescence, Cx43 is stained green with FITC, while the nucleus is dyed blue with DAPI, and the lateral Cx43 is as indicated by the arrow. (C) 2, 3, 5 – Triphenyl tetrazolium chloride staining determined myocardial infarct size. (D) Hematoxylin-Eosin staining evaluated changes in myocardial morphology. *vs group IR, #vs group N+IR. Data from n = 3 hearts. ##P<0.01, ***P<0.001.