BACKGROUND: Hypoxia/reoxygenation (H/R) is an important in vitro model for exploring the molecular mechanisms and functions of autophagy during myocardial ischemia/reperfusion (I/R). Neonatal rat cardiomyocytes (NRCM) and H9c2 cells are widely used to study H/R. METHODS: The degree of autophagy in NRCM and H9c2 cells exposed to H/R was assessed by detecting the markers of autophagy, Beclin-1 and LC3II. Autophagosomes were confirmed in both NRCM and H9c2 cells exposed to H/R using MDC staining and TEM. RESULTS: The expression levels of Beclin-1 and LC3II were significantly increased in NRCM under H/R conditions (p < 0.05 vs. control). In contrast, the expression levels of Beclin-1 and LC3II were significantly reduced in H9c2 cells exposed to H/R (p < 0.05 vs. control). The fluorescence intensity and the number of MDC-labeled particles were greater in NRCM exposed to H/R, compared to H9c2 cells (p < 0.05 vs. control). The number of autophagosomes exposed to H/R by TEM was greater in NRCM, compared to H9c2 cells, which was similar to the levels of autophagy markers observed in NRCM and H9c2 cells (p < 0.05 vs. control). CONCLUSIONS: NRCM may be more suitable to study autophagy during H/R than H9c2 cells.
BACKGROUND:Hypoxia/reoxygenation (H/R) is an important in vitro model for exploring the molecular mechanisms and functions of autophagy during myocardial ischemia/reperfusion (I/R). Neonatal rat cardiomyocytes (NRCM) and H9c2 cells are widely used to study H/R. METHODS: The degree of autophagy in NRCM and H9c2 cells exposed to H/R was assessed by detecting the markers of autophagy, Beclin-1 and LC3II. Autophagosomes were confirmed in both NRCM and H9c2 cells exposed to H/R using MDC staining and TEM. RESULTS: The expression levels of Beclin-1 and LC3II were significantly increased in NRCM under H/R conditions (p < 0.05 vs. control). In contrast, the expression levels of Beclin-1 and LC3II were significantly reduced in H9c2 cells exposed to H/R (p < 0.05 vs. control). The fluorescence intensity and the number of MDC-labeled particles were greater in NRCM exposed to H/R, compared to H9c2 cells (p < 0.05 vs. control). The number of autophagosomes exposed to H/R by TEM was greater in NRCM, compared to H9c2 cells, which was similar to the levels of autophagy markers observed in NRCM and H9c2 cells (p < 0.05 vs. control). CONCLUSIONS: NRCM may be more suitable to study autophagy during H/R than H9c2 cells.