FIGURE 6.

The myocardial oxidative stress generation and the antioxidant protein markers levels in WT, HE, and HO mice with or without moderate alcohol treatment. (A) Representative images of dihydroethidium (DHE) staining were performed to assess ROS levels in mouse heart (Images in the top row, Scale bars = 60μm); representative images of 4‐hydroxynonenal (4‐HNE) immunohistochemistry detection in mouse heart (images in the middle row, Scale bars = 80 μm). (B) Quantification of DHE staining from different visual fields of six samples in each experimental group. n = 6. (C) Measurement of the intensity of 4‐HNE immunostaining from different visual fields of six samples in each experimental group. n = 6. (D) The concentration of MDA in heart tissue homogenates, normalized to total protein amount and expressed as μmoL/mg protein. n = 4. (E) Representative western blots of HO‐1 (32 KDa) with β‐actin (43 KDa) used as loading control in mouse heart tissues. (F) Densitometry values (normalized to β‐actin) of HO‐1. n = 6. (G) Representative western blots of SOD2 (25 KDa) with β‐actin (43 KDa) used as loading control in mouse heart tissues; (H) Densitometry values (normalized to β‐actin) of SOD2. n = 6. (I) RT‐qPCR analysis of SOD2 gene in mouse heart. n = 8. The data are presented as the mean ± SEM. Statistical comparisons were carried out by one‐way ANOVA. *p < 0.05, versus WT‐con, ^# p < 0.05, versus WT‐EtOH, ^& p < 0.05, versus HE‐con, ^$ p < 0.05, versus HE‐EtOH, ^† p < 0.05, versus HO‐con