Huang Si, Wang Ya, Li Manjia, Mu Hongyu, Tan Chunlei, Wang Mingming, Zhang Feng, Sheng Jun, Tian Yang, Zhao Cunchao
College of Food Science and Technology, Yunnan Agricultural University, Kunming 650201, China.
College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.
Nutrients. 2025 Mar 12;17(6):1002. doi: 10.3390/nu17061002.
This study aims to investigate the structural and functional characteristics of walnut protein hydrolysates (WPHs) with different molecular weights prepared using protease from Dregea sinensis Hemsl, as well as the anti-fatigue effects of low-molecular-weight walnut protein hydrolysates (LWPs) and their impact on the cecal microbiota and faecal metabolism of mice. The anti-fatigue activity of WPHs with different molecular weights was evaluated, and the LWPs were analyzed in a centralized manner. A 28-day gavage study was conducted to assess LWP's anti-fatigue benefits in mice, supplemented by metabolomic analysis to explore its impact on metabolic pathways. Our findings revealed that LWP significantly outperformed unhydrolyzed walnut protein (WP) in terms of water retention, lipid retention, emulsifying properties, and foaming capacity. Notably, differential protein expression associated with LWP highlighted pathways related to antioxidant activity. In vivo studies showed that LWP markedly enhanced glycogen storage in the muscles and liver of mice, while reducing serum levels of serum urea nitrogen, lactate dehydrogenase, blood lactic acid, and creatine kinase. Furthermore, the levels of Superoxide Dismutase and Glutathione were significantly elevated, alongside a reduction in Malondialdehyde, indicating that LWP's anti-fatigue effect is closely linked to improved oxidative stress resistance. Additionally, LWP promoted beneficial increases in microbial populations such as , , , and , which are associated with enhanced fatigue resistance. Metabolomic analysis indicated significant enrichment in glycerophospholipid metabolism and amino acid biosynthesis, identifying key metabolites including palmitoylethanolamide and 4-methyl-5-thiazoleethanol, both of which are integral to health maintenance. LWP demonstrates a robust anti-fatigue effect, supported by its accessibility, straightforward preparation, and eco-friendly characteristics. These attributes suggest that LWP has promising potential for inclusion in health products aimed at enhancing vitality and combating fatigue.
本研究旨在探究用华钩藤蛋白酶制备的不同分子量核桃蛋白水解物（WPHs）的结构和功能特性，以及低分子量核桃蛋白水解物（LWPs）的抗疲劳作用及其对小鼠盲肠微生物群和粪便代谢的影响。评估了不同分子量WPHs的抗疲劳活性，并对LWPs进行了集中分析。进行了为期28天的灌胃研究，以评估LWPs对小鼠的抗疲劳益处，并辅以代谢组学分析以探索其对代谢途径的影响。我们的研究结果表明，LWPs在保水、保脂、乳化性能和发泡能力方面明显优于未水解的核桃蛋白（WP）。值得注意的是，与LWPs相关的差异蛋白表达突出了与抗氧化活性相关的途径。体内研究表明，LWPs显著增强了小鼠肌肉和肝脏中的糖原储存，同时降低了血清尿素氮、乳酸脱氢酶、血乳酸和肌酸激酶的血清水平。此外，超氧化物歧化酶和谷胱甘肽水平显著升高，同时丙二醛减少，表明LWPs的抗疲劳作用与提高抗氧化应激能力密切相关。此外，LWPs促进了与抗疲劳能力增强相关的有益微生物种群（如 、 、 和 ）的增加。代谢组学分析表明甘油磷脂代谢和氨基酸生物合成显著富集，确定了关键代谢物，包括棕榈酰乙醇胺和4-甲基-5-噻唑乙醇，这两者都是维持健康所必需的。LWPs具有强大的抗疲劳作用，其易获取、制备简单和环保的特性为其提供了支持。这些特性表明，LWPs在旨在增强活力和抗疲劳的健康产品中具有广阔的应用前景。
