Effect of  Fructus on Skeletal Muscle Mitogen Activated Protein Kinase Signaling Pathways Protein Expression and Antioxidantive Stress Damage Capacity in Over-trained Rats

CUI Xiao-mei; CAO Jian-min; ZHOU Hai-tao; WANG Can; WAN Zi-yi

doi:10.13422/j.cnki.syfjx.2017030122

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Effect of Fructus on Skeletal Muscle Mitogen Activated Protein Kinase Signaling Pathways Protein Expression and Antioxidantive Stress Damage Capacity in Over-trained Rats

更新时间：2024-01-04

- Effect of Fructus on Skeletal Muscle Mitogen Activated Protein Kinase Signaling Pathways Protein Expression and Antioxidantive Stress Damage Capacity in Over-trained Rats
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 3, Pages: 122-127(2017)
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- DOI：10.13422/j.cnki.syfjx.2017030122
  CLC： R285.5
- Published：2017
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CUI Xiao-mei, CAO Jian-min, ZHOU Hai-tao, et al. Effect of Fructus on Skeletal Muscle Mitogen Activated Protein Kinase Signaling Pathways Protein Expression and Antioxidantive Stress Damage Capacity in Over-trained Rats[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(3): 122-127.
DOI：

CUI Xiao-mei, CAO Jian-min, ZHOU Hai-tao, et al. Effect of Fructus on Skeletal Muscle Mitogen Activated Protein Kinase Signaling Pathways Protein Expression and Antioxidantive Stress Damage Capacity in Over-trained Rats[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(3): 122-127. DOI： 10.13422/j.cnki.syfjx.2017030122.

摘要

目的：研究黑果枸杞子对过度训练大鼠骨骼肌MAPK信号通道蛋白表达及抗氧化应激损伤能力的影响。方法：以等量负荷有氧运动训练和大强度耐力训练大鼠为模型，50只SPF级Wistar 49 d龄雄性大鼠为对象，随机分为4组，分别为正常组、有氧训练组、过度训练组、过度训练+黑果枸杞子组，每组12只（剔除不符合实验要求的大鼠）。每天灌胃（ig）给药1次，黑果枸杞子干预组剂量为4.48 mg·kg-1，ig体积为5 mL·kg-1，其他组ig等体积生理盐水。42 d负荷游泳训练后，测试各组大鼠骨骼肌丝裂素活化蛋白激酶（MAPK）信号系统蛋白表达及氧化应激损伤相关指标，分光光度法检测骨骼肌超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量；蛋白质免疫印迹（Western blot）法检测MAPK信号通道蛋白表达；酶联免疫吸附测试法（ELISA）检测3-硝基酪氨酸（3-NT），8-羟基脱氧鸟苷（8-OHdG）含量；乙酰半胱氨酸法检测血清肌酸激酶（CK）活性；苯肼显色法检测乳酸脱氢酶（LDH）活性。结果：力竭游泳时间，有氧训练组较正常组有所延长，黑果枸杞子干预组较过度训练组显著增加（P<0.01）。骨骼肌SOD活性，有氧训练组略高于正常组；过度训练组较正常组和有氧训练组明显下降（P<0.05），黑果枸杞子干预组较过度训练组明显升高（P<0.05）。骨骼肌MDA含量，有氧训练组略低于正常组；过度训练组较正常组和有氧训练组显著升高（P<0.01），黑果枸杞子干预组较过度训练组显著降低（P<0.01）。骨骼肌3-NT，8-OHdG，有氧训练组略高于正常组；过度训练组较正常组和有氧训练组显著升高（P<0.01）；黑果枸杞子干预组较过度训练组明显降低（P<0.05）。血清LDH活性，有氧训练组略高于正常组；过度训练组较正常组和有氧训练组均显著升高（P<0.01）；黑果枸杞子干预组较过度训练组明显升高（P<0.05）。血清CK活性，有氧训练组略高于正常组；过度训练组较正常组和有氧训练组显著升高（P<0.01）；黑果枸杞子干预组较过度训练组显著降低（P<0.01）。p38 MAPK，细胞外信号调节蛋白激酶（ERK）信号通路蛋白表达，有氧训练组高于正常组（P<0.05），低于过度训练组（P<0.05）；过度训练组高于安静组（P<0.01）；黑果枸杞子干预组高于正常组（P<0.05），低于过度训练组（P<0.05）。结论：补充黑果枸杞子可以有效缓解过度训练导致的机体内环境失衡，抑制MAPK信号通路蛋白的过度表达，激活过度训练大鼠骨骼肌MAPK信号转导系统诱导抗氧化酶基因表达，提高机体尤其是骨骼肌的氧自由基清除能力，调节机体的抗氧化应激损伤能力，预防和延缓氧化应激损伤及运动疲劳的发生与发展。

Abstract

Objective: To study the effect of Lycium ruthenicum fructus on the expression of MAPK signal channel protein and antioxidantive stress damage capacity in skeletal muscle of rats with excessive training. Method: Equivalent load aerobic training and high intensity endurance training rat model was established. A total of 50 49-day old male SPF Wistar rats were randomly divided into 4 groups

with 12 in each group (the rats that did not meet the requirements were removed):control group (C Group)

aerobic training group (M group)

overtraining group (OM group)

overtraining and Lycium ruthenicum fructus group (LM group). The dose of LM group was 4.48 mg·kg-1

with ig volume of 5 mL·kg-1. The other groups were given equal volume of saline. After 42 days' exhaustive swimming training

mitogen activated protein kinase (MAPK) signal channel protein expression and antioxidantive stress damage indicators in skeletal muscle of rats were examined. Spectrophotometry was adopted to detect activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA). Western blot was used to detect MAPK signal channel protein expression. ELISA was applied in detecting content of 3-nitrotyrosine (3-NT) and 8-hydroxy-2-deoxyguanosine (8-OHdG). Acetylcysteine method was adopted to detect activity of creatine kinase (CK). Phenylhydrazine chromogenic method was used to detect activity of lactate dehydrogenase (LDH). Result: The exhaustive swimming time of M group was longer than C group

and that of LM group was significantly longer than OM group (P<0.01). The SOD activity in M group was higher than that in C group

that in OM group was significantly lower than C group and M group (P<0.05)

and that in LM group was significantly higher than OM group (P<0.05). 3-NT and 8-OHdG content in M group was higher than that in C group

that in OM group was significantly higher than C group and M group (P<0.01)

and that in LM group was significantly lower than OM group (P<0.01). The LDH and CK activities of serum in M group was slightly higher than C group

that in OM group was significantly higher than C group and M group (P<0.01)

and that in LM group was significantly higher than OM group (P<0.05

P<0.01). The expression of p38 MAPK and ERK signal channel protein in M group was higher than that in C group (P<0.05)

and that in M group was lower than the OM group (P<0.05); that in OM group was higher than C group (P<0.01); that in LM group was higher than that in C group (P<0.05)

and that in LM group was lower than OM group (P<0.05). Conclusion: The supplementation of Lycium ruthenicum fructus can effectively alleviate the imbalance of the inner environment caused by overtraining

inhibit the excessive expression of MAPK signal channel protein

activate antioxidant enzyme gene expression induced by skeletal muscle MAPK signal transduction system of overtraining rats

improve the capacity of scavenging oxygen free radicals in body

especially skeletal muscle

regulate the antioxidantive stress damage capacity of body

and prevent and delay the oxidative stress injury and the occurrence and development of the sports fatigue.

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