摘要
目的:探讨一次性力竭运动对HIF-1α转基因鼠骨骼肌Nrf2抗氧化信号的影响。方法:HIF-1α转基因鼠和野生型C57BL/6J小鼠各20只。随机分为野生安静组(WC组)、野生运动组(WE组)、转基因安静组(HC组)和转基因运动组(HE组)。正式实验时运动组小鼠做递增负荷跑台运动直至力竭,运动后即刻取材,脱颈处死后取两侧小腿骨骼肌。采用酶联免疫法(ELISA)测定骨骼肌Nrf2-ARE结合活性,实时荧光定量PCR测定骨骼肌Nrf2下游抗氧化基因表达,Western Blot法测定骨骼肌中蛋白表达,高质荧光测定法测定小鼠骨骼肌ROS。结果:1)与WC组相比,HC组小鼠骨骼肌Nrf2 mRNA和蛋白表达显著增加,Nrf2-ARE结合活性显著增加,靶基因SOD1、SOD2、NQO-1 mRNA表达显著增加,NQO-1蛋白表达显著增加,骨骼肌ROS水平极显著下降;2)与HC组相比,HE组小鼠骨骼肌核蛋白Nrf2含量显著性增加,SOD2和NQO-1 mRNA表达量显著增加,且NQO-1蛋白表达显著增加,ROS无明显变化;3)与WE组相比,HE组小鼠Nrf2 mRNA和蛋白表达显著增加,Nrf2核蛋白及P-Nrf2表达增加。骨骼肌SOD2和NQO-1 mRNA表达增加,NQO-1蛋白表达显著增加,骨骼肌ROS含量极显著下降。结论:适当的HIF-1α高表达可提高安静状态下小鼠骨骼肌Nrf2-ARE通路的激活,且对一次力竭运动后骨骼肌ROS的消除也有积极的调节作用。
To examine the effect of HIF-1αon Nrf2-ARE antioxidant pathway in mice skeletal muscle after acute exhaustion exercise.Methods:HIF-1αhigh expression(H)and C57BL/6J mice(W)were used at 20 respectively and each kind of mice were randomly divided into two groups:control(C)and exercise(E).The treadmill exercise was preformed at the acute exhaustion exercise.On the day of acute exercise,mice allocated to perform treadmill running were subject to 5%incline and 5min at 10m/min,and then increased 3m/min every 3 minutes.Mice were sacrificed at the indicated time points following treadmill running.Nrf2,phosphor-Nrf2(Ser40),nuclear Nrf2 protein were measured by Western Blot and Nrf2-ARE binding activity,the mRNA and proetin levels of Nrf2 target genes,key antioxidant enzymes(SOD1,SOD2,CAT,NQO-1)and ROS level,were also measured in skeletal muscles after the interventions.Results:1)The results showed that compared with WC,RNA and protein expression level of Nrf2 were increased in HC skeletal muscles.Nrf2-ARE binding activity,Nrf2 target gene SOD1,SOD2,NQO-1 mRNA expression and NQO-1 protein expression were also increased in HC skeletal muscles.Meanwhile,ROS level in HC skeletal muscles decreased significantly.2)After the acute exhaustion exercise,high HIF-1αexpression mice(HE)had higher expression of p-Nrf2(Ser 40)and nuclear Nrf2 protein than the wide type mice(WE).The mRNA expression of SOD1 and mRNA/protein of NQO-1 in HE increased as well.In contrast,ROS level decreased significantly in HE muscles.Conclusion:The result indicated that the proper high expression of HIF-1αcould promote the antioxidant capacity of skeletal muscle in mice through Nrf2-ARE pathway.
作者
王林佳
张缨
WANG Lin-jia;ZHANG Ying(Beijing Sport University,Beijing 100084,China)
出处
《体育科学》
CSSCI
北大核心
2018年第11期60-65,共6页
China Sport Science
基金
国家自然科学基金资助项目(31471134)
