一次性力竭运动致大鼠骨骼肌氧化应激的机制

doi:10.12047/j.cjap.5841.2020.004

摘要/Abstract

摘要： 目的:观察一次性力竭运动对大鼠骨骼肌氧化应激相关酶表达的影响。方法:雄性SD大鼠40只,分为4组(n=10),分别为对照组(C组)、力竭运动组(E组)、运动+PKC抑制剂组(EC组)、运动+NOX抑制剂组(EA组)。三组运动大鼠进行3 d的跑台适应性运动(5 m/min,1次/日,无坡度),然后休息1 d;EC组于运动前1 d和运动前1 h注射PKC抑制剂白屈菜红碱(5 mg/kg),EA组同期注射NADPH氧化酶抑制剂Apocynin(10 mg/kg),C组和E组注射同等剂量生理盐水;三组运动大鼠进行一次性跑台力竭运动,力竭后取大鼠的跖肌,DCF荧光探针检测活性氧(ROS),Western blot分析NOX2、NOX4、3-NT,免疫沉淀分析PKC、NOX2、NOX4。结果:与C组相比,E组的ROS水平、NOX2和NOX4蛋白表达、PKC-NOX2和PKC-NOX4复合物水平、3-NT生成均显著增加(P＜0.01,P＜ 0.05),EC组、EA组ROS无显著差异(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组的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)。结论:力竭运动诱导骨骼肌NOX2、NOX4蛋白表达增加,PKC通过调控NOX2介导ROS的生成。

关键词: 力竭运动, 蛋白激酶C, NADPH氧化酶, 氧化应激, 大鼠

Abstract: 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.

Key words: exhaustive exercise, protein kinase C, NADPH oxidase, oxidative stress, rat

刘姣, 周刚, 梅雨, 谢文杰, 李鹏飞, 杨帆. 一次性力竭运动致大鼠骨骼肌氧化应激的机制[J]. 中国应用生理学杂志, 2020, 36(1): 17-22.

LIU Jiao, ZHOU Gang, MEI Yu, XIE Wen-jie, LI Peng-fei, YANG Fan. Mechanism of oxidative stress in skeletal muscle of rats induced by acute exhaustive exercise[J]. CJAP, 2020, 36(1): 17-22.

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