The effect of ozone treatment on extremity in rats with acute limb ischemia master's thesis in medicine

Objective: Lower extremity ischemia is one of the major causes of mortality and morbidity presenting as a multifactorial presentation which is frequently seen in our country as acute or chronic. The reperfusion injury caused by hypoxia and revascularization during ischemia is reversible and irreversible. It has been shown that controlled ozone administration will provide protection against the oxidative stress that will occur during ischemia and reperfusion, or stimulate adaptation, prevent damage caused by reactive oxygen species and protect against ischemia-reperfusion (I / R) damage. In this study, we are planning to investigate the effect of ozone therapy on the extremities in rats with lower extremity ischemia. Materials and Methods: In this study, 21 Wistar Albino rats with an average weight of 200-400 g were randomly divided into 3 groups. Only subcutaneous incision was made to the ratsin the control group (n = 5). Ischemia-reperfusion (IR) group (n = 8) was applied to theischemia by clamping the infrarenal abdominal aorta for 60 min in rats. After the clampswere removed, reperfusion was performed for 120 min. Ischemiareperfusion (IR) + treatment (n = 8) rats were started 4 days before the surgical procedure and 20 mcg / ml medical ozone was administered intraperitoneally daily. After the ozone treatment was completed, ischemia was induced by clamping the infrarenal abdominal aorta for 60 min in rats. After the clamps were removed, reperfusion was performed for 120 min. After the reperfusion period was over, all rats, including the control group, were sacrificed and appropriate blood samples were taken. Gastrocnemius muscle tissue was removed for biochemical evaluation and stored under appropriate conditions. The investigations were performed with serum in muscle tissue and especially the changes related to prooxidant-antioxidant system and inflammation were investigated. In this context, serum ROS, MDA, AOPP, GSH, FRAP, TNFα, NRF2, SOD and GSH-Px levels were determined. In addition, lipid peroxide and protein oxidation products, antioxidant system elements, tumor necrosis factor-alpha (TNF-α) and nuclear factor erythroid 2 related factor 2 (NRF2) were determined in the tissues. Result: In biochemical investigations; no change was observed in serum ROS, MDA, AOPP, GSH and FRAP, TNFα, NRF2 levels, SOD and GSH-Px activities. In the studies on muscle tissue oxidant system; ROS, MDA and AOPP levels were increased with ischemia-reperfusion. A significant decrease was observed in these parameters with medical ozone application. GSH and SOD levels were significantly decreased in the muscle tissue antioxidant system. Ozone (O3) treatment showed a significant increase in these values. As a result of I / R injury, no significant changes were observed in TNF-a levels in muscle tissue and a significant increase in NRF2 was observed. No changes were observed in the TNF-α and NRF2 results with the application of ozone. Conclusion: In conclusion, we can say that in IR group there is ischemiareperfusion injury in lower extremity and ozone treatment is protective against this injury. Key Words: Antioxidant, Ischemia-reperfusion, ozone