Cucurbitacin I Protects H9c2 Cardiomyoblasts against H₂O₂-Induced Oxidative Stress via Protection of Mitochondrial Dysfunction

Figure 2

Cu I prevents the mortality and accumulation of ROS production in H₂O₂-treated H9c2 cardiomyoblasts. (a) Cell viability was measured using the MTT assay in cells pretreated with 0.1, 0.5, and 1 μM Cu I for 24 h followed by exposure to 500 μM H₂O₂ for additional 24 h. ROS production was assessed by DCFH-DA staining. (b) Representative images and (c) fluorescence intensities in cells pretreated with 0.1, 0.5, and 1 μM Cu I for 24 h followed by exposure to 500 μM H₂O₂ for additional 24 h. (d) Western blot analysis of SOD-1, catalase, and GPx protein expression levels. (e) Protein expression levels were quantified by scanning densitometry. β-Actin was used as the loading control. Western blot analysis was performed in triplicate with three independent samples. Data are expressed as fold changes ± SEM versus control cells. Significance was analyzed by a one-way ANOVA followed by the Bonferroni post hoc test. and versus control cells. , , and versus H₂O₂ alone-treated cells. Cont: control; Cu I: cucurbitacin I. Scale bar: 100 μm.

(a)

(b)

(c)

(d)

(e)