研究简报 SHORT COMMUNICATION

电刺激频率对肉鸡成肌细胞抗氧化能力及核因子E2相关因子2/抗氧化反应序列元件信号通路中抗氧化相关基因表达的影响

  • 胥蕾 ,
  • 张欣 ,
  • 黄华云 ,
  • 李宁 ,
  • 万晓莉 ,
  • 杨海明 ,
  • 王志跃
展开
  • 1. 扬州大学动物科学与技术学院, 扬州 225009;
    2. 江苏省家禽科学研究所, 扬州 225125
胥蕾(1984-),女,四川雅安人,讲师,博士,从事家禽营养与生产研究。E-mail:xlei@yzu.edu.cn

收稿日期: 2020-04-18

  网络出版日期: 2020-11-16

基金资助

国家自然科学基金项目(31601978);2018年江苏省"双创计划"科技副总项目(FZ20180271);江苏高校优势学科建设工程资助项目(PAPD)

Effects of Electrical Stimulation Frequencies on Antioxidant Capacity and Expression of Antioxidant-Related Genes in Factor-Erythroid 2-Related Factor 2/Antioxidant Reaction Sequence Element Signaling Pathway in Broiler Myoblasts

  • XU Lei ,
  • ZHANG Xin ,
  • HUANG Huayun ,
  • LI Ning ,
  • WAN Xiaoli ,
  • YANG Haiming ,
  • WANG Zhiyue
Expand
  • 1. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;
    2. Jiangsu Institute of Poultry Science, Yangzhou 225125, China

Received date: 2020-04-18

  Online published: 2020-11-16

摘要

本试验旨在研究不同的电刺激频率对肉鸡成肌细胞抗氧化能力及核因子E2相关因子2(Nrf2)/抗氧化反应序列元件(ARE)信号通路中抗氧化相关基因表达的影响。试验将分化3 d的肉鸡成肌细胞分为对照组和5个电刺激组,每组3个重复。对照组不进行电刺激处理,处理组的电刺激频率分别为50、160、320、640和1 280 Hz,刺激电压和时间统一为30 V和10 s。电刺激处理后继续培养30 min,测定肉鸡成肌细胞抗氧化能力和Nrf2/ARE信号通路中抗氧化相关基因表达的变化。结果表明:不同的电刺激频率对细胞损伤程度无显著影响(P>0.05);320 Hz组细胞过氧化氢(H2O2)含量显著低于50、640和1 280 Hz组(P<0.05),还原型谷胱甘肽(GSH)含量和超氧化物歧化酶(SOD)活性显著高于对照组和640 Hz组(P<0.05);320 Hz组成肌细胞中Nrf2、Kelch样环氧氯丙烷相关蛋白-1(Keap1)和NAD (P)H醌氧还原酶1(NQO1)的mRNA相对表达量显著高于其他电刺激组(P<0.05),谷胱甘肽-S-转移酶T1(GSTT1)的mRNA相对表达量最高,与对照组和其他电刺激组均存在显著差异(P<0.05)。综上所述,在电刺激30 V和10 s的情况下,320 Hz的电刺激频率可以提高肉鸡成肌细胞的抗氧化能力和Nrf2/ARE信号通路中部分抗氧化相关基因的mRNA相对表达量,但并没有改变细胞形态或引起明显的细胞膜破损。

本文引用格式

胥蕾 , 张欣 , 黄华云 , 李宁 , 万晓莉 , 杨海明 , 王志跃 . 电刺激频率对肉鸡成肌细胞抗氧化能力及核因子E2相关因子2/抗氧化反应序列元件信号通路中抗氧化相关基因表达的影响[J]. 动物营养学报, 2020 , 32(11) : 5415 -5423 . DOI: 10.3969/j.issn.1006-267x.2020.11.048

Abstract

This experiment was to investigate the effects of different electrical stimulation frequencies on the antioxidant capacity and expression of antioxidant-related genes in factor-erythroid 2-related factor 2 (Nrf2)/antioxidant reaction sequence element (ARE) signaling pathway in broiler myoblasts. The myoblasts were divided into the control group and five electrical stimulation groups, with three replicates in each group. The myoblasts in control group were not treated with electrical stimulation. The myoblasts in the other groups were respectively stimulated with 50, 160, 320, 640 and 1 280 Hz electricity combined with a stable voltage of 30 V and a stimulation period of 10 s. Cells were cultured for 30 min after the treatments. Then the changes of antioxidant capacity and expression of antioxidant-related genes in Nrf2/ARE signaling pathway were measured in broiler myoblasts. The results showed that different electrical frequencies had no significant effect on the degree of cell injury (P>0.05). The hydrogen peroxide (H2O2) content in 320 Hz group was significantly lower than that in 50, 640 and 1 280 Hz groups (P<0.05). The reduced glutathione (GSH)content and superoxide dismutase (SOD) activity in 320 Hz group were significantly higher than those in the control group and 640 Hz group (P<0.05). The mRNA relative expression levels of Nrf2, Kelch-like ECH-associated protein 1 (Keap1) and NAD(P)H quinone oxidoreductase l (NQO1) were significantly higher in 320 Hz group than those in other electrical stimulation groups (P<0.05). The mRNA relative expression of glutathione-S-transferase T1 (GSTT1) was the highest in 320 Hz group as compared with other groups (P<0.05). In conclusion, a stimulating frequency at 320 Hz can improve the antioxidant capacity and the expression of antioxidant-related genes in Nrf2/ARE signaling pathway without changing cell morphology or causing obvious damage to cell membrane in broiler myoblasts.

参考文献

[1] 黄继超,王鹏,徐幸莲,等.肉鸡宰前击晕研究进展[J].食品科学,2013,34(11):344-347.
[2] XU L,ZHANG H J,YUE H Y,et al.Low-current & high-frequency electrical stunning increased oxidative stress,lipid peroxidation,and gene transcription of the mitogen-activated protein kinase/nuclear factor-erythroid 2-related factor 2/antioxidant responsive element (MAPK/Nrf2/ARE) signaling pathway in breast muscle of broilers[J].Food Chemistry,2018,242:491-496.
[3] 胥蕾.致晕方法影响肉仔鸡肉品质的机理及脂质过氧化调控[D].博士学位论文.北京:中国农业科学院,2011.
[4] XU L,ZHANG H J,WAN X L,et al.The evaluation of pre-slaughter low-current/high-frequency ES on lipid oxidative stability and antioxidant status in thigh muscle of broilers[J].International Journal of Food Science and Technology,2019,doi:10.1111/ijfs.14402.
[5] KREGEL K C,ZHANG H J.An integrated view of oxidative stress in aging:basic mechanisms,functional effects,and pathological considerations[J].American Journal of Physiology.Regulatory,Integrative and Comparative Physiology,2007,292(1):R18-R36.
[6] FINKEL T.Signal transduction by reactive oxygen species[J].Journal of Cell Biology,2011,194(1):7-15.  
[7] 李婷婷.京海黄鸡成肌分化关键mRNAs,IncRNAs和miRNAs筛选[D].博士学位论文.扬州:扬州大学,2018.
[8] FROST R A,NYSTROM G J,LANG C H.Tumor necrosis factor-α decreases insulin-like growth factor-Ⅰ messenger ribonucleic acid expression in C2C12 myoblasts via a Jun N-terminal kinase pathway[J].Endocrinology,2003,144(5):1770-1779.  
[9] 董云峰,赵秀峰,徐晓阳.电刺激对C2C12肌管自由基代谢及Nrf2/ARE信号通路的影响[J].中国运动医学杂志,2017,36(12):1066-1071.
[10] 刘永,潘其乐,罗冬梅.电刺激对体外培养C2C12细胞抗氧化能力的影响[J].北京体育大学学报,2014,37(5):67-71.
[11] LUO W,WU H,YE Y,et al.The transient expression of miR-203 and its inhibiting effects on skeletal muscle cell proliferation and differentiation[J].Cell Death and Disease,2014,5(7):e1347.
[12] 刘小利,李慧娟.电刺激对软骨细胞增殖及细胞内Ca2+浓度的影响[J].山西医科大学学报,2018,49(8):912-916.
[13] 魏严.体外培养心肌细胞对力、电刺激的生物学响应的研究[D].硕士学位论文.北京:中国人民解放军军事医学科学院,2008.
[14] LOBNER D.Comparison of the LDH and MTT assays for quantifying cell death:validity for neuronal apoptosis?[J].Journal of Neuroscience Methods,2000,96(2):147-152.  
[15] 潘红英.电刺激C2C12收缩运动对自由基代谢影响的实验研究[D].硕士学位论文.广州:华南师范大学,2005.
[16] IRRCHER I,HOOD D A.Regulation of Egr-1,SRF,and Sp1 mRNA expression in contracting skeletal muscle cells[J].Journal of Applied Physiology,2004,97(6):2207-2213.  
[17] ZHANG J K,YANG L,MENG G L,et al.Protective effect of tetrahydroxystilbene glucoside against hydrogen peroxide-induced dysfunction and oxidative stress in osteoblastic MC3T3-E1 cells[J].European Journal of Pharmacology,2012,689(1/2/3):31-37.
[18] 吴志红.Nrf2-ARE通路在奶牛乳腺上皮细胞氧化损伤中的作用研究[D].硕士学位论文.呼和浩特:内蒙古农业大学,2017.
[19] RAMOS-GOMEZ M,KWAK M K,DOLAN P M,et al.Sensitivity to carcinogenesis is increased and chemoprotective efficacy of enzyme inducers is lost in Nrf2 transcription factor-deficient mice[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98(6):3410-3415.  
[20] HASAN W N W,KWAK M K,MAKPOL S,et al.Piper betle induces phase Ⅰ & Ⅱ genes through Nrf2/ARE signaling pathway in mouse embryonic fibroblasts derived from wild type and Nrf2 knockout cells[J].BMC Complementary and Alternative Medicine,2014,14:72.
[21] 张倩.金丝桃苷对Nrf2-ARE途径的影响及其对肝细胞氧化应激损伤的保护作用[D].硕士学位论文.重庆:西南大学,2014.
[22] BREWER A C,MURRAY T V A,ARNO M,et al.Nox4 regulates Nrf2 and glutathione redox in cardiomyocytes in vivo[J].Free Radical Biology and Medicine,2011,51(1):205-215.  
[23] 杨丽梦,熊旭东.Keap1-Nrf2/ARE氧化应激信号通路与脓毒症急性肺损伤的相关性研究[J].临床急诊杂志,2016(8):590-592.
[24] 王芳,刘健,叶英法,等.基于Keap1-Nrf2/ARE信号传导通路探讨黄芪多糖改善干燥综合征模型大鼠心功能的机制[J].中国中西医结合杂志,2014,34(5):566-574.
[25] 董合玲,吴洪渊,唐钰,等.AMPK调控不同强度电刺激引起的C2C12肌管线粒体氧化应激[J].南方医科大学学报,2018,38(6):742-747.
[26] LO VERSO F,CARNIO S,VAINSHTEIN A,et al.Autophagy is not required to sustain exercise and PRKAA1/AMPK activity but is important to prevent mitochondrial damage during physical activity[J].Autophagy,2014,10(11):1883-1894.  
[27] 齐晓龙.共轭亚油酸对产蛋鸡抗氧化机能的影响[D].博士学位论文.北京:中国农业科学院,2013.
文章导航

/