The effect of aerobic exercise training and Octapamine supplementation on FOXO3 protein and ATP Synthase gene expression in the cardiac muscle of rats fed with deep-fried oil

The use of repeatedly heated oils has adverse effects on human health, causing oxidative stress and dysfunction of genes and proteins. Oxidative stress disrupts the regulation of vascular walls and heart cells, mitochondrial damage and cellular apoptosis in heart cells. Myocardial cells, especially mitochondria, become vulnerable to oxidative stress and the ability to their reproduction and regeneration are reduced. The two proteins involved in the process of mitochondrial biogenesis of the cardiac muscle and free radicals in the cell are FOXO3 and ATP Synthase proteins. FOXO3 is like a double-edged sword and on the other hand prevents pathological myocardial hypertrophy, which maintains the size of cardiomyocytes by inhibiting the growth of cardiomyocytes and promoting autophagy. On the other hand, in conditions of ROS accumulation, it promotes cell and mitochondria towards apoptosis. ATP Synthase is associated with octopamine because of its role in relaxing the cardiac muscle. Octopamine (an endogenous biogenic amine) has been introduced as a plant extract with anti-anxiety, sedative, anticonvulsant effects and promotes the growth and growth of nerve stem cells. Octopamine, which is naturally present in many plants such as oranges, has positive effects. In controlling heart diseases such as controlling the nervous system of the heart, reducing tachycardia, and relaxing the cardiac muscle. This study aimed to evaluate the effect of aerobic exercise training and octopamine on FOXO3 protein and ATP Synthase gene expression in the cardiac muscle of rats fed deep-fried oils.

30 8-week-old Wistar rats in 5 groups (6) including healthy control, consumption of deep-fried oils, consumption of deep-fried oils + aerobic exercise, consumption of deep-fried oils + supplements Octapamine, consumption of deep-fried oils + aerobic exercise + octopamine supplement were divided, during the study period, deep heated oils were fed orally (gavage, 10ml / kg) to the tested rats for 4 weeks. (Morning). Deep-fried oil was obtained from 8 liters of sunflower oil, which was heated at 190 to 200 ° C for 8 consecutive days. According to sources every 30 minutes. Foods including chicken nuggets, potatoes, and protein products (sausages and sausages) were immersed in oil and at the end of the fourth day, the oil was stored for use in the poisoning intervention. DFO oil was orally gavaged to rats for 4 weeks 5 days a week (10 ml/kg). The supplement used in this study was octopamine. The duration of complementary intervention was 4 weeks and 5 days per week. The dose used according to the articles was 81 μmol / kg by intraperitoneal injection (IP solution with 9% normal saline). The training protocol was performed with moderate intensity training in the range of VO2 max 65-50%, which includes 5 sessions of 30 minutes per week (treadmill) with 5 minutes of warm-up, 20 minutes of activity, and 5 minutes of cooling down. On the first day of training, the speed starts from 16 m / s and increases according to the protocol every week, and reaches 26 m / s on the last day after 4 weeks. 48 hours after the last intervention with at least 8 hours of fasting with chloroform solution anesthetized after splitting the chest, a blood sample was taken from the left ventricle of the heart with a 3 cc syringe. The collected blood was placed in a simple 12 ml tubes and the EDTA tube was placed in a refrigerated centrifuge to collect serum and plasma. After blood sampling from the heart, the tissues were quickly isolated and the tissue was washed with saline phosphate buffer solution (PBS) and placed in the microtube. Alterations of FOXO3 protein expression and ATP Synthase gene expression by Western blotting by Primary and secondary antibodies and Real-time-PCR were performed by specific primers. To determine the effect of deep-frying oil intake, healthy control group, and control group - deep-fried oil were compared using an independent t-test. A two-way analysis of variance was used to determine the main effect of exercise, the main effect of octopamine, and the interaction between exercise and octopamine. If there was a significant difference, the Bonferroni post hoc test was used to determine the location of the difference. The expression of the desired gene was calculated by the formula 2∆∆ct and the values of multiplication changes.

Compared to the control group who did not receive supplements and exercise significantly reduced the expression of FOXO3 protein in the exercise group (P = 0.005). Octopamine supplementation also significantly reduced FOXO3 protein expression (P = 0.001). The combination of exercise and supplements was also significant compared to the control group (P = 0.001). Although the lowest expression of FOXO3 protein was observed in the exercise group and octopamine supplementation, the interaction of these two interventions was not statistically significant (P = 0.159). (Figure 2 Section A and Table 1). Compared to the control group who did not receive supplements and exercise, exercise significantly increased the expression of the ATP Synthase gene (P = 0.012). Octopamine supplementation also significantly increased ATP Synthase gene expression (P = 0.001). The combination of exercise and supplements was also significant compared to the control group (P = 0.0001). Although the highest expression of ATP Synthase gene belonged to the exercise group and octopamine supplementation, the interaction between exercise and octopamine expression of this gene was not statistically significant. (P = 0.071).

It has been reported that moderate-intensity exercise training can reduce myocardial infarction during cell myocardial infarction in rats by modifying FOX family proteins, enhance cardiovascular function, autophagy and Promote the degradation of damaged proteins. Octopamine as a lipolytic agent probably acts on rat fat cells and it is conceivable that octopamine to increase Stem cell proliferation rate is effective and mimics some of the effects of noradrenaline and adrenaline. This response probably indicates a positive and antioxidant effect, anti-inflammatory, weight loss, fat burning, and anti-cancer effects of octopamine, which has been able to act on cells damaged by heated oil poisoning and the damage caused by DFO. According to the results of the present study, it seems that aerobic exercise training and octopamine can reduce the destructive effects of deep heated oils. Octopamine probably plays a very important role in regulating autophagy and mitochondrial function as an antioxidant and aerobic exercise, so exercise can be a key process in cellular and molecular mechanisms.