Background & Aims: Free radicals and reactive oxygen species (ROS), especially superoxide anion and hydroxyl radical, play significant roles in various pulmonary diseases. The interaction between superoxide and nitric oxide potentially produces harmful and strong oxidants such as peroxynitrite. Due to continuous exposure to environmental oxygen and exogenous free radicals, lungs are susceptible to ROS-induced damage caused by cigarette smoke or air pollutants. Hydrogen peroxide (H₂O₂) is a mild and relatively stable oxidant widely used as a marker of ROS to evaluate cellular responses to oxidative stress and is produced during oxidative processes. To protect against the harmful effects of ROS, an antioxidant system exists in the lung that includes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). These enzymes are endogenously produced and are known as enzymatic antioxidants. SOD converts superoxide anion to hydrogen peroxide (H₂O₂), and catalase converts hydrogen peroxide into water and oxygen. Besides enzymatic antioxidants, other antioxidants such as vitamins exist, known as non-enzymatic antioxidants, among which vitamin D is included. Previous studies have shown that consumption of antioxidant-rich foods such as vitamins A and E is associated with reduced pulmonary-related diseases. Vitamin D, a fat-soluble steroid hormone, can be produced under the skin by ultraviolet sunlight exposure or obtained through dietary intake. Vitamin D deficiency leads to mild oxidative stress and increased proteolysis in muscles, and based on these studies, it is recognized as a potential antioxidant. Physical inactivity is one of the major global challenges in the 21st century. Exercise induces ROS formation, acting as an important mediator of physiological signaling, cellular adaptations, muscle contraction modulation, antioxidant defense regulation, and oxidative damage repair. Aerobic exercise transiently increases ROS production but reduces oxidative stress-related diseases, and regular aerobic exercise over the long term protects the body against oxidative stress. Evidence indicates that exercise can enhance antioxidant activity in the heart, muscle, and lung. Numerous studies have investigated the effects of aerobic exercise on oxidative stress and antioxidant activity, although results have been inconsistent. Moreover, limited research has focused specifically on lung tissue. Therefore, the present study aimed to investigate the effects of aerobic exercise combined with vitamin D on the concentration of SOD and CAT in lung tissue of rats exposed to hydrogen peroxide. Methods: In an experimental trial, 30 adult male Wistar rats weighing 220±20 g and aged 8 to 10 weeks were obtained from the animal center of Shiraz University. Rats were housed in polycarbonate cages (5 rats per cage) at 22±2°C temperature, 55% humidity, and a 12:12-hour light-dark cycle with free access to food and water. All animal procedures were performed according to ethical guidelines and approved by the Research Deputy of Kerman University of Medical Sciences. Rats were acclimated for two weeks before protocol initiation and then randomly divided into five groups (6 rats per group): hydrogen peroxide group, hydrogen peroxide + vitamin D group, hydrogen peroxide + aerobic exercise group, and hydrogen peroxide + aerobic exercise + vitamin D group. Aerobic exercise groups were familiarized with treadmill running for 10 days. This study was approved by the Ethics Committee with code IR.KMU.REC.1396.1562 from the Ministry of Health and Medical Education, Kerman University of Medical Sciences and Health Services, Research and Technology Center. To induce oxidative stress, all rats received intraperitoneal injections of hydrogen peroxide (Merck, Germany) at 0.1 mg/kg body weight, three times weekly on alternate days. Rats undergoing aerobic exercise ran daily on a treadmill for 8 weeks. During week 1, rats exercised at 8 m/min speed and 10° incline for 30 minutes. In week 2, speed increased to 12 m/min with the same incline and duration. Week 3 involved running at 16 m/min for 45 minutes. In week 4, exercise was performed at 20 m/min and similar incline for 45 minutes. From weeks 5 to 8, rats ran daily at 20 m/min with a 10° incline for 60 minutes. Vitamin D3 treatment was administered as intraperitoneal injections of 1 µg/day for 8 weeks using DITHRECOL (Caspian Vitamin, 300,000 IU/ml). Lung tissue SOD and CAT concentrations were measured by ELISA. Data were analyzed by two-way ANOVA followed by Bonferroni post hoc test. Results: Interaction of aerobic exercise and vitamin D significantly increased lung SOD and CAT concentrations (p<0.05). Conclusion: Overall, the interaction of aerobic exercise and vitamin D significantly enhanced lung SOD and CAT levels, indicating the effectiveness of aerobic exercise combined with vitamin D3 supplementation compared to inactivity in improving antioxidant status in lung tissue. Study limitations include the lack of measurement of ROS and total antioxidant capacity (TAC) in lung tissue. Given increasing air pollution and oxidative stress-related lung diseases, future studies are recommended to investigate the effects of combined aerobic exercise and vitamin D3 on apoptosis in lung tissue. |