Study on the Antioxidant and Anti-fatigue Effect of Wheat Peptides
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摘要: 目的：探讨小麦肽的体外抗氧化活性和体内抗疲劳作用。方法：通过H2O2诱导小鼠成纤维细胞L929构建氧化应激损伤模型，测定损伤后L929细胞的存活率、乳酸脱氢酶（lactic dehydrogenase, LDH）、超氧化物歧化酶（superoxide dismutase, SOD）和谷胱甘肽过氧化物酶（glutathione peroxide dismutase, GSH-Px）活力以及丙二醛（MDA）含量，从细胞水平评价小麦肽的抗氧化作用。然后通过力竭游泳和自由泳实验，测定力竭游泳时间、乳酸（lactic acid, LA）、尿素氮（blood urea nitrogen, BUN）和肌糖原（muscle glycogen, MG）含量、SOD和GSH-Px活力以及MDA含量，来评价小麦肽的体内抗疲劳和抗氧化作用。结果：小麦肽浓度在0.4~0.8 mg/mL时对L929细胞的H2O2损伤有极显著的保护作用（P<0.01)；与模型组相比，小麦肽浓度为0.6和0.8 mg/mL时的LDH活力分别显著下降了20.79%和19.67%（P<0.05)，SOD活力分别显著（P<0.05)和极显著（P<0.01)提高了83.21%和95.19%，GSH-Px活力分别提高了28.69%和32.14%，MDA含量分别显著下降了25.91%和26.99%（P<0.01) 。体内抗疲劳实验表明，与对照组相比，2个剂量组的小鼠力竭游泳时间分别极显著延长了72.93%和91.73%（P<0.01)，LA含量分别极显著下降了24.65%和25.16%（P<0.01)，BUN含量分别极显著（P<0.01)和显著（P<0.05)下降了19.74%和17.78%，MG含量分别极显著提高了48.63%和56.85%（P<0.01)；体内抗氧化结果表明，2个剂量组的SOD和 GSH-Px活力分别极显著提高了20.54%、25.91%和29.79%、35.77%（P<0.01)，MDA含量分别极显著下降了23.08%和21.46%（P<0.01)。Pearson相关性分析表明小麦肽的体内抗疲劳与抗氧化作用高度相关。结论：小麦肽具有显著的抗氧化和缓解疲劳作用，且抗氧化活性与抗疲劳作用高度相关。Abstract: Objective: To investigate the in vitro antioxidant activity and in vivo anti-fatigue effect of wheat peptides. Methods: The oxidative stress model was established by treating L929 cells with H2O2. Then the cell survival rate, the activity of lactic dehydrogenase (LDH), superoxide dismutase (SOD) and glutathione peroxide dismutase (GSH-Px), and the content of malondialdehyde (MDA) were measured to evaluate the antioxidant activity of wheat peptides in vitro. In addition, the exhaustive swimming and freestyle swimming test was assessed. Then the exhaustive swimming time, the content of lactic acid (LA), blood urea nitrogen (BUN), muscle glycogen (MG) and MDA, the activity of SOD and GSH-Px were measured to study the anti-fatigue and antioxidant of wheat peptides in vivo. Results: The wheat peptides in the concentration range from 0.4 to 0.8 mg/mL could protect L929 cells against H2O2-induced oxidative damage by highly significantly increasing the survival rate（P<0.01). Compared to the model group, the activity of LDH of the wheat peptides at 0.6 and 0.8 mg/mL significantly decreased by 20.79% and 19.67%（P<0.05) , the activity of SOD was significantly （P<0.05) and highly significantly（P<0.01) increased by 83.21% and 95.19%, the activity of GSH-Px increased by 28.69% and 32.14%, and the content of MDA significantly decreased by 25.91% and 26.99%（P<0.05). Compared with the control group, the exhaustive swimming time of the wheat peptides at 2 and 8 mg/mL was significantly increased by 72.93% and 91.73%（P<0.01) , the content of LA significantly decreased by 24.65% and 25.16%（P<0.01) , the content of BUN highly significantly （P<0.01) and significantly（P<0.05) decreased by 19.74% and 17.78%, and the content of MG highly significantly increased by 48.63% and 56.85%（P<0.01) . At the same time, the activity of SOD and GSH-Px was highly significantly increased by 20.54% and 25.91%, 29.79% and 35.77%（P<0.01) , and the content of MDA was highly significantly decreased by 23.08% and 21.46%（P<0.01) . Pearson correlation analysis showed that the antifatigue effect of wheat peptides was highly correlated with its antioxidant activity. Conclusion: Wheat peptide had significant antioxidant activity and anti-fatigue effect, and the anti-fatigue effect of wheat peptide was correlated with antioxidant activity.
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Keywords:
- wheat peptides /
- mice /
- antioxidant /
- anti-fatigue /
- free radical
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图 2 小麦肽对L929细胞氧化损伤的保护作用
注：与对照组相比，*代表差异显著P<0.05，**代表差异极显著P<0.01；与模型组相比，#代表差异显著P<0.05，##代表差异极显著P<0.01，图3同。
Figure 2. Protective effect of wheat peptides against H2O2 induced oxidative damage in L929 cells
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图 3 小麦肽对L929细胞氧化损伤LDH、SOD、GSH-Px和MDA水平的影响
Figure 3. Effects of wheat peptides against H2O2 induced oxidative damage in L929 cells on the SOD, GSH-Px, MDA and LDH
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图 4 小麦肽对小鼠力竭游泳时间、乳酸、尿素氮和肌糖原水平的影响
注：与对照组相比，*代表差异显著P<0.05，**代表差异极显著P<0.01，图5同。
Figure 4. Effect of wheat peptides on the exhaustive swimming time, LA, BUN and MG
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图 5 小麦肽对小鼠SOD、GSH-Px和MDA水平的影响
Figure 5. Effect of wheat peptides on the SOD, GSH-Px and MDA
表 1 动物实验设计
Table 1 Experimental design of animals
组别 灌胃物 剂量（mg/g） 小鼠数量（只） 对照 蒸馏水 同等体积 16 阳性 乳清蛋白 2 16 低剂量 小麦肽 2 16 高剂量 小麦肽 8 16 
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表 2 小麦肽的氨基酸组成
Table 2 Amino acid composition of wheat peptides
氨基酸种类 含量（g/100 g） 氨基酸种类 含量（g/100 g） 天冬氨酸 1.76 酪氨酸 3.46 谷氨酸 4.61 缬氨酸 3.98 谷氨酰胺 22.91 甲硫氨酸 1.46 丝氨酸 2.55 半胱氨酸 0.39 甘氨酸 2.83 异亮氨酸 3.81 组氨酸 1.00 亮氨酸 5.43 精氨酸 3.36 苯丙氨酸 4.25 苏氨酸 1.59 赖氨酸 1.12 丙氨酸 2.23 总量 76.66 脯氨酸 9.92
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表 3 小麦肽的相对分子质量分布
Table 3 Relative molecular weight distribution of wheat peptides
分子量（Da） >10000 5000~10000 3000~5000 180~3000 <180 峰面积比例（%） 13.35 11.46 4.63 62.71 7.85
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表 4 小麦肽对小鼠体重的影响
Table 4 Effect of wheat peptides on body weight in mice
组别 体重（g） 开始 结束 增重 对照 25.52±1.10 32.70±2.22 7.18±1.78 阳性 25.76±1.03 34.03±2.65 8.27±1.86 低剂量 25.63±0.69 33.65±1.53 8.02±1.19 高剂量 24.92±1.26 33.46±3.06 8.54±2.28
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表 5 体内抗氧化活性与缓解疲劳作用的Pearson相关系数
Table 5 Pearson correlation analysis between antioxidant activity and anti-fatigue activities in vivo
项目 力竭游泳时间 LA含量 BUN含量 MG含量 SOD活力 0.986* −0.976* −0.961* 0.986* GSH-Px活力 0.992** −0.984* −0.969* 0.978* MDA含量 −0.966* 0.996** 0.999** −0.939 注：*代表差异显著，P<0.05，**代表差异极显著，P<0.01。
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