Effect of Allopurinol Pretreatment on Superoxide Dismutase Activity in Ischemia Reperfusion Injury to Skeletal Muscles of the Hindlimbs of the Rats

Shin, Kye Cheol; Kim, Dong Hun; Paek, Doo Jin; Chung, Ho Sam

Abstract

It has been well known that ischemia reperfusion injury to skeletal muscle following an acute arterial occulusion causes significant morbidity and mortality. The skeletal muscle, which contains high energy phosphate compounds, has ischemic tolerance. During the ischemia, the ATP is catalyzed to hypoxanthine anaerobically and hypoxanthine dehydrogenase is converted to xanthine oxidase. During reperfusion, the hypoxanthine is catalyzed to xanthine by xanthine oxidase under O2 presence and that results in production of cytotoxic oxygen free radicals. The authors perform the present study to investigate the effects of allopurinol, the inhibitor of xanthine oxidase, on reperfused ischemic skeletal muscles by measuring of the immunoreactivities and exzyme activities of superoxide dismutase(SOD) and the formation of malondialdehyde(MDA). A total of 104 healthy Sprague-Dawley rats weighting from 200 gm to 250 gm were used as experimental animals. Under urethane(3.0mg/kg., IP) anesthesia with 3.0mg/kg of urethane, lower abdominal incision was made and the right and left common iliac artery were ligated by using vascular clamp for 2 hours. Both the quandriceps femoris muscles were obtained at 0 hour, half hour, 1 hour, 3 hours, 6 hours and 12 hours after the removal of vascular clamp. In the allopurinol pretreated group, 50mg/kg of allopurinol was administered once a day for 2 days and before 2 hours of ischemia. The specimens were sectioned in 14µm thickness with cryostat and homogenated ischemia. The specimens were sectioned in 14 µm thickness with cryostat and homegenated in the phosphate buffer. The immunoreactivities and enzyme activities of SOD were observed. The results were as follows: 1. The immunoreactivitiy and enzyme activity of SOD are decreased and the MDA level is increased in the 2 hours inchemic quadriceps femoris muscle of rats. 2. During the reperfusion of ischemic quadriceps femoris muscle of rats, the immunoreactivities of SOD in the half hour reperfused ischemic group and the enzyme activities of SOD in the 1 hour reperfused group are the highest and the immunoreactivities and enzyme activities of 6 hours reperfused ischemic group are the lowest. 3. Pertreatment of allopurionl decreased the immunoreactivities and enzyme activities of SOD during the ischemia and reperfusion of the quadriceps femoris muscles of rat. This results suggest that the allopurinol decreases the damages of skeletal muscles of rate during ischemia and reperfusion.