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中国应用生理学杂志 ›› 2021, Vol. 37 ›› Issue (6): 688-693.doi: 10.12047/j.cjap.6160.2021.079

• 研究论文 • 上一篇    下一篇

游泳训练对小鼠心肌PKCδ/P66shc蛋白表达的影响*

谢文杰, 周刚Δ, 李鹏飞, 杨帆, 安静芳, 李航   

  1. 湖南大学体育学院, 长沙 410082
  • 收稿日期:2020-09-19 修回日期:2021-02-01 出版日期:2021-11-28 发布日期:2021-11-25
  • 通讯作者: Tel: 13548644844; E-mail: zg460@126.com
  • 基金资助:
    *湖南省自然科学基金项目(12JJ3093)

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

XIE Wen-jie, ZHOU GangΔ, LI Peng-fei, YANG Fan, AN Jing-fang, LI Hang   

  1. College of Physical Education, Hunan University, Changsha 410000, China
  • Received:2020-09-19 Revised:2021-02-01 Online:2021-11-28 Published:2021-11-25

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

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