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[急性力竭运动诱导大鼠骨骼肌氧化应激的机制]

[Mechanism of oxidative stress in skeletal muscle of rats induced by acute exhaustive exercise].

作者信息

Liu Jiao, Zhou Gang, Mei Yu, Xie Wen-Jie, Li Peng-Fei, Yang Fan

机构信息

College of Physical Education, Hunan University, Changsha 410082, China.

出版信息

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Jan 28;36(1):17-22. doi: 10.12047/j.cjap.5841.2020.004.

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.

摘要

目的

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。

方法

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。

结果

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)。

结论

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。

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