Effects Of Fatigue On NADPH Oxidase, Ros And Apoptosis Via20-HETE And PKC In Rat Cardiomyocytes

Fatigue refers to the organism could not be at the level of a particular function of maintaining or not maintaining the preset intensity and the function temporarily weakened.20-Hydroxyeicosatetraenoic acid (20-HETE) is a small molecule active compound of arachidonic acid catalyzed by the cytochrome P450ω-hydroxylase20-HETE plays an important role in regulating cardiovascular function.To reveals rat myocardial cell in fatigue whether produces and release20-HETE, whether activates PKC, and effects NADPH oxidase-derived ROS production, induces myocardial cell apoptosis, we established fatigue rat model through swimming endurance training, and these rats were divided into four groups:controlled group, sports fatigue group, sports fatigue+20-HETE inhibitors HET0016group and sports fatigue+PKC inhibitors chelerythrine (CHE) group.This paper studies from these aspects:1) Organization NADPH oxidase activity analysis with chemiluminescence detection, found that fatigue significantly enhances NADPH oxidase activity of rats in the control group rats up to177.77%; intraperitoneal injection of20-HETE inhibitors HET0016to exercise-induced fatigue rats, NADPH oxidase activity diminished the fatigue group24%, reveals the fatigue generated by promoting20-HETE, enhances the oxidative injury of myocardial cells; intraperitoneal injection of PKC inhibitors chelerythrine of exercise-induced fatigue in rats and found it more NADPH OXIDASE reduces the fatigue group52%, reveals a fatigue activation of PKC, enhancing myocardial oxidative injury.2) ROS fluorescent probe-Ⅱ hydrogen b piperidine (Dihydroethidium, DHE), laser total focused software determination the Group produces of red fluorescent, found that fatigue enhances ROS activity38.267%than controlled group; celiac injection20-HETE inhibitors HET0016of fatigue group, found its ROS activity has weakened17.07%than fatigue group, reveals has fatigue through promotion20-HETE generated, Enhanced myocardial oxidative injury; intraperitoneal injection of PKC inhibitors chelerythrine of exercise-induced fatigue rats, had discovered ROS activity down from fatigue Group11.047%, reveals a fatigue activation of PKC, enhancing myocardial oxidative injury.3) Caspase-3Spectrophotometric method for testing, found that compared with the control group, the activity of activated apoptosis protein Caspase-3in sports fatigue increase up76.7%, reveal fatigue contributed to the myocardial cell apoptosis; exercise-induced fatigue+20-HETE inhibitor HET0016group, found that compared with the fatigue group, its Caspase-3activity reduces24.1%, Reveal fatigue generated by promoting20-HETE, enhances myocardial cell apoptosis; fatigue+chelerythrine Group, found that, compared with the Group of exercise-induced fatigue, which significantly reduces Caspase-3activity15.6%. Reveal fatigue activate PKC, induction of apoptosis in myocardial cells. These results reveal that sports fatigue produces and release endogenous substances20-HETE, activates PKC and enhances NADPH oxidase-derived ROS production, activates apoptosis protein Caspase-3activity and inducts myocardial cells apoptosis. This provides a theoretical basis for further perfect the mechanism of the exercise-induced fatigue on heart theory.