Department of Nutrition and Food Hygiene, School of Public Health, Peking University, Beijing 100191, China.
Nutrients. 2021 Jun 19;13(6):2100. doi: 10.3390/nu13062100.
This study investigated the antioxidant effects of whey protein peptide on learning and memory in aging C57BL/6N mice. A total of 72 SPF male C57BL/6N mice were used. Twelve mice were randomly selected as the control group, and the other mice were intraperitoneally injected with D-galactose (100 mg/kg body weight for 6 weeks), during which, the mice in the control group were intraperitoneally injected with the same amount of normal saline. After 6 weeks, the blood was taken from the epicanthus and the serum MDA level was measured, according to which, the mice were randomly divided into the model control group, the whey protein group (1.5 g/kg body weight), and three Whey protein peptide (WHP) intervention groups (0.3 g/kg body weight, 1.5 g/kg body weight, 3.0 g/kg body weight). The water solution of the test sample was administered by oral gavage every day. The intervention period was 30 days, during which, the model control group, the whey protein group, and the whey protein peptide group continued receiving intraperitoneal injections of D-galactose, while the control group continued receiving intraperitoneal injections of normal saline. After the intervention, behavioral experiments were conducted in the following order: open field test, water maze test, and new object recognition test. After the behavioral experiment, the morphology of hippocampal formation was observed by HE staining and TUNEL labeling. Oxidative stress-related indexes in the serum, liver, and brain were detected. Expression levels of the cholinergic system-related enzymes and proinflammatory cytokines in brain tissue were detected. Western blot was used to detect the expression of synaptic plasticity-related proteins in the mouse brain. The results showed that WHP could significantly improve the accumulation of MDA and PC, increase the activities of SOD and GSH-Px, resist oxidative stress injury, and enhance the potential of endogenous antioxidant defense mechanisms. WHP can significantly improve the decline of aging-related spatial exploration, body movement, and spatial and non-spatial learning/memory ability. Its specific mechanism may be related to reducing the degeneration of hippocampal nerve cells, reducing the apoptosis of nerve cells, improving the activity of AChE, reducing the expression of inflammatory factors (TNF-α and IL-1β) in brain tissue, reducing oxidative stress injury, and improving the expression of p-CaMKⅡ and BDNF synaptic plasticity protein. These results indicate that WHP can improve aging-related oxidative stress, as well as learning and memory impairment.
本研究探讨了乳清蛋白肽对衰老 C57BL/6N 小鼠学习记忆的抗氧化作用。使用了 72 只 SPF 雄性 C57BL/6N 小鼠。随机选择 12 只作为对照组,其余小鼠腹腔注射 D-半乳糖(100mg/kg 体重,连续 6 周),对照组小鼠同时腹腔注射等量生理盐水。6 周后,从内眦采血,测定血清 MDA 水平,根据 MDA 水平将小鼠随机分为模型对照组、乳清蛋白组(1.5g/kg 体重)和三个乳清蛋白肽(WHP)干预组(0.3g/kg 体重、1.5g/kg 体重、3.0g/kg 体重)。每天通过灌胃给予测试样品的水溶液。干预期为 30 天,在此期间,模型对照组、乳清蛋白组和乳清蛋白肽组继续腹腔注射 D-半乳糖,对照组继续腹腔注射生理盐水。干预后,按以下顺序进行行为学实验:旷场实验、水迷宫实验和新物体识别实验。行为学实验后,采用 HE 染色和 TUNEL 标记观察海马结构形态。检测血清、肝、脑组织中氧化应激相关指标。检测脑组织中胆碱能系统相关酶和促炎细胞因子的表达水平。采用 Western blot 检测小鼠脑组织中突触可塑性相关蛋白的表达。结果表明,WHP 可显著改善 MDA 和 PC 的堆积,提高 SOD 和 GSH-Px 的活性,抵抗氧化应激损伤,增强内源性抗氧化防御机制的潜能。WHP 能显著改善与衰老相关的空间探索、身体运动以及空间和非空间学习/记忆能力的下降。其具体机制可能与减轻海马神经细胞的变性、减少神经细胞的凋亡、提高 AChE 的活性、降低脑组织中炎症因子(TNF-α和 IL-1β)的表达、减轻氧化应激损伤以及提高 p-CaMKⅡ和 BDNF 突触可塑性蛋白的表达有关。这些结果表明,WHP 可改善与衰老相关的氧化应激以及学习记忆障碍。