Objective: To observe the effects of acute exhaustive exercise on the expressions of oxidative stress related enzymes in skeletal muscle of rats.
Methods: Forty male SD rats were divided into 4 groups, 10 rats in each group, which were the control group (C group), exhausted exercise group (E group), exercise + PKC inhibitor group (EC group), exercise + NOX inhibitor group (EA group). Three groups of exercise rats were familiarized with treadmill running for 3 days (5 m/min, once/d, no incline), then rested for one day. EC group was injected with PKC inhibitor chelerythrine (5 mg / kg) one day before and one hour before exercise, EA group was injected with NADPH oxidase inhibitor apocynin (10 mg / kg) at the same time, group C and group E were injected with the same dose of normal saline. Three groups of exercise rats were subjected to a one-time treadmill exhaustion exercise, and the plantaris were taken after exhaustion. Reactive oxygen species (ROS) were detected by DCF fluorescent probe, NOX2, NOX4, 3-NT were analyzed by Western blot, and PKC, NOX2, NOX4 were analyzed by immunoprecipitation.
Results: Compared with group C, ROS level, NOX2 and NOX4 protein expressions, PKC-NOX2 and PKC-NOX4 complex levels, and 3-NT production in group E were significantly increased (P＜0.01, P＜0.05), and ROS level was no significant difference in group EC and group EA (P＞0.05), and NOX4 protein expression in group EC was significantly increased (P ＜ 0.05). Compared with group E, ROS level, NOX2 and NOX4 protein expressions, PKC-NOX2 and PKC-NOX4 complex levels, 3-NT production were decreased significantly (P＜0.01, P＜0.05).
Conclusion: Exhausted exercise induces increased expressions of NOX2 and NOX4 proteins in skeletal muscle, and PKC mediates the production of ROS by regulating NOX2.