游泳训练对小鼠心肌PKCδ/P66shc蛋白表达的影响*

doi:10.12047/j.cjap.6160.2021.079

摘要/Abstract

摘要： 目的:探究不同强度的游泳训练对小鼠心肌P66shc蛋白的影响。方法:将50只昆明小鼠随机分为对照组(C组)、负重游泳组(E组)、负重游泳+药物组(ER组)、非负重游泳组(P组)、非负重游泳+药物组(PR组),10只/组。C组不运动,E组、ER组、P组、PR组进行4周游泳训练,其中E组、ER组以体重3%负荷进行负重游泳,P组、PR组无负重游泳,60 min/d,每周6次。ER组、PR组小鼠在最后2次运动前腹腔注射PKCδ抑制剂Rottlerin(0.3 mg/kg),C组、E组、P组注射同等剂量生理盐水。在训练结束24 h后取样,Western blot测定小鼠心肌PKCδ、P-PKCδ、P66shc、P-P66shc、NOX2蛋白表达;免疫共沉淀测PKCδ和P66shc;生化分析心肌及血清丙二醛(MDA)、心肌活性氧(ROS)、超氧化物歧化酶(SOD)。结果:与C组比较,E组的PKCδ、P-PKCδ、P66shc、P-P66shc、NOX2蛋白表达均明显增加(P＜0.01),血清和心肌MDA水平、心肌ROS明显增加(P＜0.05或P＜0.01),心肌SOD活性降低(P＜0.01),P组的PKCδ、P-PKCδ、P-P66shc和NOX2明显增加(P＜0.05或P＜0.01),心肌SOD活性增强(P＜0.05);与E组比较,ER组PKCδ(P＜0.01)、P-PKCδ(P＜0.01)、P66shc(P＜0.05)、P-P66shc(P＜0.01)、NOX2(P＜0.05)蛋白表达明显减少,P组P66shc蛋白表达显著减少(P＜0.05),心肌MDA(P＜0.01)和ROS(P＜ 0.05)减少,SOD活性增强(P＜0.01);与P组比较,PR组的PKCδ、P-PKCδ、P-P66shc蛋白表达明显减少(P＜ 0.01),NOX2增加(P＜0.05)。结论:两种强度的游泳训练均促使小鼠心肌细胞内PKCδ蛋白及其磷酸化增加;高强度游泳训练可显著增强P66shc蛋白表达及磷酸化水平,导致ROS大量生成,抗氧化酶活性下降;低强度游泳训练增强P66shc磷酸化但不促进其蛋白表达,心肌抗氧化能力增强,产生运动适应。

关键词: 蛋白激酶Cδ, 游泳训练, P66shc, 心肌, 活性氧, 小鼠

Abstract: Objective: To investigate the effects of different intensity of swimming training on p66Shc protein in mouse myocardium. Methods: Fifty Kunming mice were randomly divided into control group (Group C), weight-bearing swimming group (Group E), weight-bearing swimming + drug group (Group ER), non weight-bearing swimming group (Group P), non weight-bearing swimming + drug group (Group PR), with 10 mice in each group. Group C did not exercise. Groups E, ER, P, and PR received swimming training for 4 weeks. Groups E and ER performed weight-bearing swimming with a 3% body weight, and Group P and Group PR were swimming without weight-bearing, 60 min/d, 6 times/w. Mice in ER and PR groups were injected intraperitoneally with Rottlerin (0.3 mg/kg), a PKCδ inhibitor, before the last two exercises. Groups C, E, and P were injected with the same dose of normal saline. Samples were collected after training finished for 24 hours. The protein expressions of PKCδ, P-PKCδ, P66Shc, P-P66shc and NOX2 were detected by Western blot; PKCδ and P66Shc were detected by immunoprecipitation; malondialdehyde (MDA), reactive oxygen species (ROS) and superoxide dismutase (SOD) in myocardium and serum were analyzed by biochemistry. Results: Compared with Group C, the protein expressions of PKCδ, P-PKCδ, P66Shc, P-P66shc and NOX2 in Group E were increased significantly (P＜ 0.01), the serum and myocardial MDA levels, myocardial ROS were increased significantly (P＜0.05 or P＜0.01), and the myocardial SOD activity was decreased (P＜0.01), the PKCδ, P-PKCδ, P-P66shc and NOX2 in Group P were increased significantly (P＜0.05 or P＜0.01), and the myocardial SOD activity was enhanced (P＜0.05). Compared with Group E, the protein expressionS of PKCδ (P＜0.01), P-PKCδ (P＜0.01), P66Shc (P＜0.05), P-P66shc (P＜0.01), NOX2 (P＜0.05) in Group ER was decreased significantly, the protein expression of P66Shc in Group P was decreased significantly (P＜0.05), the myocardial MDA (P＜0.01) and ROS (P＜0.05) were decreased, and the activity of SOD was enhanced (P＜0.01). Compared with Group P, the protein expressions of PKCδ, P-PKCδ and P-P66shc in Group PR were decreased significantly (P＜0.01), while the expression of NOX2 was increased (P＜0.05). Conclusion: Both swimming training of two intensities promoted the increase of PKCδ protein and its phosphorylation in mouse cardiomyocytes. High-intensity swimming training could significantly enhance the expression and phosphorylation level of p66Shc protein, resulting in the production of ROS and the decrease of antioxidant enzyme activity. Low-intensity swimming training enhanced the phosphorylation of p66Shc, but did not promote its protein expression, resulting in the enhancement of myocardial antioxidant capacity and exercise adaptation.

Key words: protein kinase Cδ, swimming training, P66shc, myocardium, reactive oxygen species, mouse

中图分类号:

G804.7

谢文杰, 周刚, 李鹏飞, 杨帆, 安静芳, 李航. 游泳训练对小鼠心肌PKCδ/P66shc蛋白表达的影响^*[J]. 中国应用生理学杂志, 2021, 37(6): 688-693.

XIE Wen-jie, ZHOU Gang, LI Peng-fei, YANG Fan, AN Jing-fang, LI Hang. Effect of swimming training on the expression of PKC δ/p66Shc protein in mouse myocardium[J]. CJAP, 2021, 37(6): 688-693.

参考文献

[1] Pinho CA, Tromm CB, Tavares AMV, et al. Effects of different physical training protocols on ventricular oxidative stress parameters in infarction-induced rats[J]. Life Sci, 2012, 90(13-14): 553-559.
[2] Powers SK, Quindry JC, Kavazis AN. Exercise-induced cardioprotection against myocardial ischemia-reperfusion injury[J]. Free Radic Biol Med, 2008, 44(2): 193-201.
[3] Quintanilha AT, Packer L, Vitamin E. Physical exercise and tissue oxidative damage[J]. Ciba Found Symp, 1983, 101: 56-69.
[4] Bejma J, Ramires P, Ji LL. Free radical generation and oxidative stress with ageing and exercise: differential effects in the myocardium and liver[J]. Acta Physiol Scand, 2010, 169(4): 343-351.
[5] Jackson MJ, Vasilaki A, McArdle A. Cellular mechanisms underlying oxidative stress in human exercise[J]. Free Radic Biol Med, 2016, 98: 13-17.
[6] Krylatov AV, Maslov LN, Voronkov NS, et al. Reactive oxygen species as intracellular signaling molecules in the cardiovascular system[J]. Curr Cardiol Rev, 2018, 14(4): 290-300.
[7] Ziolkowski W, Flis DJ, Halon M, et al. Prolonged swimming promotes cellular oxidative stress and p66Shc phosphorylation, but does not induce oxidative stress in mitochondria in the rat heart[J]. Free Radic Res, 2015, 49(1): 7-16.
[8] Morita M, Matsuzaki H, Yamamoto T, et al. Epidermal growth factor receptor phosphorylates protein kinase Cδ at Tyr332 to form a trimeric complex with p66Shc in the H2O2-stimulated Cells[J]. J Biochem, 2008, 143(1): 31-8.
[9] Wang P, Li CG, Qi ZT, et al. Acute exercise induced mitochondrial H2O2 production in mouse skeletal muscle: association with p(66Shc) and FOXO3a signaling and antioxidant enzymes[J]. Oxid Med Cell Longev, 2015, ID536456: 1-10.
[10] Steinberg, SF. Structural basis of protein kinase C isoform function[J]. Physiol Rev, 2008, 88(4): 1341-1378.
[11] Giorgio M, Migliaccio E, Orsini F, et al. Electron transfer between cytochrome C and p66Shc generates reactive oxygen species that trigger mitochondrial apoptosis[J]. Cell, 2005, 122(2): 221-233.
[12] Migliaccio E, Giorgio M, Mele S, et al. The p66shc adaptor protein controls oxidative stress response and life span in mammals[J]. Nature, 1999, 402: 309-313.
[13] 马治云. 急性力竭运动对大鼠心肌PKCα、δ和ε表达的影响[J]. 中国应用生理学杂志, 2016, 32(4): 333-335.
[14] 刘姣, 周刚, 梅雨, 等. 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.
[15] Lv P, Miao SB, Shu YN, et al. Phosphorylation of smooth muscle 22α facilitates angiotensin II-Induced ROS production via activation of the PKCδ-P47phox axis through release of PKCδ and actin dynamics and is associated with hypertrophy and hyperplasia of vascular smooth muscle cell[J]. Circ Res, 2012, 111(6): 697-707.
[16] Nemoto S. Redox regulation of forkhead proteins through a p66shc-dependent signaling pathway[J]. Science, 2002, 295(5564): 2450-2452.
[17] Francia P, Cosentino F, Schiavoni M, et al. p66Shc protein, oxidative stress, and cardiovascular complications of diabetes: the missing link[J]. J Mol Med (Berl), 2009, 87(9): 885-891.
[18] Powers SK, Jackson MJ. Exercise-induced oxidative stress: cellular mechanisms and impact on muscle force production[J]. Physiol Rev, 2008, 88(4): 1243-1276.
[19] Jackson MJ, McArdle A. Role of reactive oxygen species in age-related neuromuscular deficits[J]. J Physiol, 2016, 594(8): 1979-1988.

游泳训练对小鼠心肌PKCδ/P66shc蛋白表达的影响^*

Effect of swimming training on the expression of PKC δ/p66Shc protein in mouse myocardium

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	白林林, 王志华, 宁学聪, 巩翠珂, 李爱敏, 段玉玲, 焦雨佼. IL-17A对慢性阻塞性肺疾病的干预作用及其机制^*[J]. 中国应用生理学杂志, 2021, 37(6): 584-588.
[2]	薛峰, 王辉, 许思多, 刘首挺, 龚永生, 范小芳. 左旋卡尼汀对脂多糖诱导小鼠肺微血管内皮细胞自噬和凋亡的影响^*[J]. 中国应用生理学杂志, 2021, 37(6): 589-593.
[3]	李莉, 黄涵柽. 莱菔子乙醇提取物对ApoE^-/-小鼠血脂血糖及肝脂肪变性的影响^*[J]. 中国应用生理学杂志, 2021, 37(6): 622-627.
[4]	胡佳楠, 姜辉, 李美琦, 朱俐, 骆倩倩. 模拟高原低氧环境对小鼠脾脏铁代谢的影响^*[J]. 中国应用生理学杂志, 2021, 37(6): 628-631.
[5]	李萍, 梁琳琅, 王宇, 侯达, 杨昕, 杨博. 二甲双胍和西格列汀对胰岛素抵抗糖尿病前期KKAy小鼠胰岛β细胞功能的影响^*[J]. 中国应用生理学杂志, 2021, 37(6): 644-649.
[6]	袁前发, 何珏, 徐志忠, 林多朵, 朱敬敬, 温春燕, 段海珍, 王文强. 重复经颅磁刺激联合低剂量氟西汀对CUMS抑郁小鼠的影响^*[J]. 中国应用生理学杂志, 2021, 37(6): 650-653.
[7]	吴江立, 王晓影, 代成, 李睿嘉, 张越, 孙佳欢, 李爱英. 归脾汤对大鼠心肌缺血的干预作用^*[J]. 中国应用生理学杂志, 2021, 37(6): 694-698.
[8]	孟香红, 曾斌, 陈慧玲, 李伟东, 李娜, 徐小勇. 改良法分离培养SD新生乳鼠原代心肌细胞^*[J]. 中国应用生理学杂志, 2021, 37(6): 699-704.
[9]	王小琴, 石卉, 方小霞, 彭聿平. α1-肾上腺素受体对胶原诱导性关节炎小鼠Treg细胞功能的影响^*[J]. 中国应用生理学杂志, 2021, 37(5): 449-453.
[10]	陈旭娥, 马婷, 盛良, 王文雯, 李季. IL-21单克隆抗体对MRL/lpr狼疮小鼠的治疗作用^*[J]. 中国应用生理学杂志, 2021, 37(5): 467-470.
[11]	贺英昊, 方情, 杨玲玲, 陈思敏, 骆燕萍, 钱虹, 齐敏友. 酮康唑对小鼠肝脏和睾丸生理功能的影响^*[J]. 中国应用生理学杂志, 2021, 37(5): 475-479.
[12]	臧茂林, 余梦迪, 陈重华, 黄梦琪, 罗鹏, 樊红琨, 杨春. 腹主动脉缩窄小鼠心脏结构和功能的动态变化^*[J]. 中国应用生理学杂志, 2021, 37(5): 479-482.
[13]	赵士博, 黄锁义, 潘智, 彭晓敏, 彭晓靖, 伍世娟, 韦德智, 陈新鹏. 薏苡叶和淫羊藿配伍对小鼠抗疲劳及耐缺氧的作用^*[J]. 中国应用生理学杂志, 2021, 37(5): 510-513.
[14]	朱明了, 郑珍, 楼恩哲, 赵凯婷, 何施燕, 陈佳玉. 大蒜阿霍烯抗胃癌的作用及分子机制^*[J]. 中国应用生理学杂志, 2021, 37(5): 514-519.
[15]	陈奕帆, 张李君, 刘清华, 李学文. 西格列汀对糖尿病小鼠心肌重构和自噬的影响及其机制[J]. 中国应用生理学杂志, 2021, 37(5): 534-537.