银耳多糖对力竭运动小鼠氧化性损伤所致疲劳的缓解作用

黄小兰; 黄少波; 邵丽

doi:10.13386/j.issn1002-0306.2023040074

银耳多糖对力竭运动小鼠氧化性损伤所致疲劳的缓解作用

Effect of Tremella fuciformis Polysaccharides on Fatigue Induced by Oxidative Damage in Exhaustive Exercise Mice

摘要

摘要: 目的：探讨银耳多糖（Tremella fuciformis polysaccharides，TFP）对小鼠力竭运动性氧化损伤的影响，并分析其机制。方法：采用TFP处理L6细胞48 h，CCK-8法检测L6的活力；L6细胞设置3组，包括对照组、H₂O₂组、H₂O₂+TFP组，孵育48 h，生化分析仪检测乳酸（Lactic Acid，LA）水平，Western blot实验检测Nrf2、NQO1和HO-1蛋白水平；将C57BL/6小鼠随机分为4组（每组n=10），包括模型组、模型+TFP组（50、100、200 mg/kg），模型+TFP组连续灌胃TFP，模型组灌胃相同剂量的蒸馏水，每天1次，连续2周。末次给药30 min后进行力竭运动，记录力竭游泳运动时间，采用酶联免疫吸附试验（ELISA）检测肝糖原（LG）、骨骼肌糖原（MG）；ELISA法检测血尿素氮（BUN）、LA、ROS、丙二醛（MDA）、超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）、TNF-α、IL-6、NF-κBp65水平。Western blot实验检测Nrf2、NQO1、HO-1蛋白水平。结果：H₂O₂组L6细胞的LA水平极显著（P<0.01）高于对照组，Nrf2、NQO1、HO-1蛋白水平极显著（P<0.01）低于对照组，TFP极显著（P<0.01）降低培养基中LA水平，极显著（P<0.01）上调L6细胞中Nrf2、NQO1、HO-1表达。TFP显著延长力竭游泳时间（P<0.05，P<0.01），极显著降低LA（P<0.01）、BUN（P<0.05，P<0.01）水平，显著增加肝糖原（P<0.05，P<0.01）和肌糖原（P<0.05，P<0.01）水平，极显著上调SOD（P<0.01）、GSH-Px（P<0.01），显著下调MDA（P<0.05，P<0.01）、ROS（P<0.01）水平，极显著（P<0.01）降低血清中TNF-α、IL-6、NF-κBp65的水平；与模型组比较，TFP显著增加Nrf2（P<0.05，P<0.01）、HO-1（P<0.05，P<0.01）、NQO-1（P<0.05，P<0.01）蛋白表达水平。结论：TFP通过抑制氧化应激、炎症改善力竭游泳运动引起的氧化损伤，其机制与调节Nrf2/HO-1信号通路相关。

Abstract: Objective: To investigate the effect of Tremella fuciformis polysaccharides (TFP) on oxidative damage induced by exhaustive exercise in mice and analyze its mechanism. Methods: L6 cells were treated with TFP for 48 hours, and the activity of L6 was detected by CCK-8 method. L6 cells were divided into three groups, including the control, H₂O₂, and H₂O₂+TFP groups, and incubated for 48 hours. LA levels were measured by a biochemical analyzer, and Nrf2, NQO1, and HO-1 protein levels were detected by Western blot assay. C57BL/6 mice were randomly divided into four groups (n=10 for each group), including model group and model+TFP group (50, 100, 200 mg/kg). The model+TFP groups were continuously gavaged with TFP, while the model group was gavaged with the same dose of distilled water once a day for two consecutive weeks. Exhaustive exercise was performed 30 minutes after the last administration, the duration of exhaustive swimming exercise was recorded, and enzyme-linked immunosorbent assay (ELISA) was used to detect liver glycogen (LG) and skeletal muscle glycogen (MG). ELISA was used to detect blood urea nitrogen (BUN), lactic acid (LA), ROS, malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), TNF-α, IL-6 and NF-κBp65 levels. Western blot assay was used to detect the levels of Nrf2, NQO1, and HO-1 proteins. Results: The LA levels of L6 cells in the H₂O₂ group were significantly (P<0.01) higher than those in the control group, while the protein levels of Nrf2, NQO1, and HO-1 were significantly (P<0.01) lower than those in the control group. TFP could significantly (P<0.01) reduce the LA levels in the culture medium and upregulate the expression of Nrf2, NQO1, HO-1 in L6 cells. TFP could significantly prolong exhaustive swimming time (P<0.01), reduce LA (P<0.01) and BUN (P<0.05, P<0.01) levels, increase liver and muscle glycogen levels (P<0.05, P<0.01), upregulate SOD (P<0.01) and GSH-Px (P<0.01), downregulate MDA (P<0.05, P<0.01) and ROS (P<0.01) levels, and reduce TNF-α (P<0.01), IL-6 (P<0.01) and NF-κBp65 (P<0.01). Compared with the model group, TFP could significantly increase Nrf2 (P<0.05, P<0.01), HO-1 (P<0.05, P<0.01) and NQO-1 (P<0.05, P<0.01) protein expression levels. Conclusion: TFP improved oxidative damage caused by exhaustive swimming by inhibiting oxidative stress and inflammation, and its mechanism was related to regulating the Nrf2/HO-1 signaling pathway.