Plant extract and chemicals Ginsenodie-Rg1 (Rg1, molecular weight 801.01, Figure 1) was obtained from the NuLiv Science USA, Inc, Walnut,

CA, USA. All the other chemicals used in this study were obtained from Sigma Chemicals (St. Louis, MO, USA) and Cayman Chemical Company (Ann Arbor, MI, USA). Figure 1 Chemical structure of ginsenoside-Rg1. Grouping and treatment Weight matched rats were equally divided into control (N = 20) and Rg1 (N = 20) groups. Rg1 was dissolved in 0.9% saline, and administered to Rg1 group daily at the dose of 0.1 mg/kg body weight (b.w) by gastric gavage for 10 weeks. Similarly, control group rats received the same amount of saline for the same duration. Exercise protocol In this study, rats performed swimming until exhaustion in a water pool. The water temperature was maintained https://www.selleckchem.com/ferroptosis.html at 33 ± 1°C. Three days prior to acute Temsirolimus exhaustive swimming challenge, all animals were familiarized with swimming environment for 10 min/day. Then, half number of rats (N = 10) from each group were performed an exhaustive swimming with a lead ingot (3% body weight) loaded to the tail of each rat. Rats were swimming

until exhaustion and clearly monitored to avoid sink in the pool. The swimming duration was not significantly different between control and Rg1 groups. Tissue collection Immediately after exhaustive exercise, rats were anesthetized with chloral hydrate injection (400 mg/kg b.w., intraperitoneally). The tibialis anterior (TA) muscle from the hind limbs of exercised and non-exercised rats were quickly excised and frozen ADAMTS5 into liquid nitrogen, and then stored at −80°C until biochemical analyses. 100 mg of muscle tissue was homogenized in 1 mL of Tris buffer (50 mM, pH 7.5) and centrifuged at 10000 g for 10 min at 4°C. Collected supernatant was used for the estimation of protein carbonyl (PC) and glutathione

levels. The same supernatant was also used to measure the activities of catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST) and xanthine oxidase (XO). Determination of lipid and protein oxidation Lipid peroxidation marker malondialdehyde (MDA) in muscle samples was measured spectrophotometrically as described by Ohkawa et al. [16]. Muscle tissue was homogenized in phosphate buffer (50 mM, pH 7.0) and centrifuged at 10000 g for 10 min at 4°C. This assay is based on the MDA-TBA (thiobarbituric acid) compound formed by the reaction between MDA and TBA at high temperature (90-100°C). The MDA-TBA was quantified at 450 nm by spectrophotometer. Protein oxidation in the muscle samples was determined by measuring the protein carbonyl residues by using the DNPH (2,4-dinitrophenylhydrazine).