Liu Jiao, Zhou Gang, Mei Yu, Xie Wen-Jie, Li Peng-Fei, Yang Fan
College of Physical Education, Hunan University, Changsha 410082, China.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Jan 28;36(1):17-22. doi: 10.12047/j.cjap.5841.2020.004.
To observe the effects of acute exhaustive exercise on the expressions of oxidative stress related enzymes in skeletal muscle of rats.
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.
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).
Exhausted exercise induces increased expressions of NOX2 and NOX4 proteins in skeletal muscle, and PKC mediates the production of ROS by regulating NOX2.
观察急性力竭运动对大鼠骨骼肌氧化应激相关酶表达的影响。
将40只雄性SD大鼠分为4组,每组10只,即对照组(C组)、力竭运动组(E组)、运动+蛋白激酶C(PKC)抑制剂组(EC组)、运动+烟酰胺腺嘌呤二核苷酸磷酸氧化酶(NOX)抑制剂组(EA组)。3组运动大鼠进行3天的跑步机跑步适应性训练(5米/分钟,每天1次,无坡度),然后休息1天。EC组在运动前1天和运动前1小时注射PKC抑制剂白屈菜红碱(5毫克/千克),EA组同时注射NADPH氧化酶抑制剂白杨素(10毫克/千克),C组和E组注射相同剂量的生理盐水。3组运动大鼠进行一次性跑步机力竭运动,运动后取腓肠肌。采用2',7'-二氯二氢荧光素(DCF)荧光探针检测活性氧(ROS),蛋白质免疫印迹法分析NOX2、NOX4、3-硝基酪氨酸(3-NT),免疫沉淀法分析PKC、NOX2、NOX4。
与C组相比,E组ROS水平、NOX2和NOX4蛋白表达、PKC-NOX2和PKC-NOX4复合物水平以及3-NT生成均显著增加(P<0.01,P<0.05),EC组和EA组ROS水平差异无统计学意义(P>0.05),EC组NOX4蛋白表达显著增加(P<0.05)。与E组相比,EC组和EA组ROS水平、NOX2和NOX4蛋白表达、PKC-NOX2和PKC-NOX4复合物水平、3-NT生成均显著降低(P<0.01,P<0.05)。
力竭运动可诱导大鼠骨骼肌中NOX2和NOX4蛋白表达增加,PKC通过调节NOX2介导ROS的产生。