Dtba 100

The hearts were harvested from the rats and washed with normal saline. Subsequently, 0.5 g of the ischemic heart tissue was ground at 0–4°C, following which the myocardial homogenate was centrifuged at 2,500 × g for 30 min. The supernatants (~0.4 g) were harvested and stored at −80°C until MDA concentration and SOD activity assays were performed. A thiobarbituric acid reactive substance assay (DTBA-100; Bioassay Systems LLC, Hayward, CA, USA) was used to measure the levels of MDA, by measuring the absorbance value at a wavelength of 532 nm using a microplate reader (SP-Max 2300A; Molecular Devices, LLC, Sunnyvale, CA, USA). A xanthine oxide method was used to determine SOD activity, by measuring the absorbance value at a wavelength of 550 nm. An MDA Assay kit and an SOD Assay kit (Nanjing Jiancheng Bioengineering Co., Ltd., Nanjing, China) were used to measure MDA concentration and SOD activity, respectively, according to the manufacturer's protocol. The conditions of the assays were as follows: MDA, 90–100°C for 50 min; SOD, 37°C for 20 min.