A fluorescent probe dihydroethidium (DHE) (Servicebio, Wuhan, China) was used to detect the production of ROS in myocardial tissue. DHE staining solution was added into frozen sections and the slices were incubated at 37°C for 30 min. The treated sections were then washed with PBS three times for 5 min each and incubated with DAPI solution in a dark environment for 10 min. Finally, the sections were observed using a fluorescence microscope (Eclipse C1, Nikon, Tokyo, Japan).
The heart tissue samples were homogenized into a 10% (W/V) homogenate and obtained by centrifugation at 3,500 rpm for 10 min. Biochemical analyses were performed using the supernatant. Levels of SOD (A001-1), MDA (A003-1), GSH (A006-2) and CAT (A007-2) were obtained by commercially available kits (JianCheng, Nanjing, China), respectively, on the basis of the manufacturer’s instructions.
The heart tissue samples were homogenized into a 10% (W/V) homogenate and obtained by centrifugation at 3,500 rpm for 10 min. Biochemical analyses were performed using the supernatant. Levels of SOD (A001-1), MDA (A003-1), GSH (A006-2) and CAT (A007-2) were obtained by commercially available kits (JianCheng, Nanjing, China), respectively, on the basis of the manufacturer’s instructions.