College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China.
Department of Pharmacy, Wuhan City Central Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430014, China.
J Ethnopharmacol. 2021 Jun 28;274:114047. doi: 10.1016/j.jep.2021.114047. Epub 2021 Mar 19.
For many centuries, Mauremys mutica is highly valued as a food homologous Chinese herbal medicine. It has been considered useful to sedate, nourish brain and promote sleep. However, the animal experimental evidence of its sleep-promoting activity is missing in literature.
In this study, PCPA-induced insomnia model was used to explore the sleep-promoting mechanism of enzymolysis peptides from PMM, and its main composition and chemical structure were analyzed.
Experiments were performed using PCPA-induced insomnia model, all animals were intraperitoneally injected with PCPA (350 mg/kg·d) for two days. The sleep-promoting effect evaluated using measuring content of 5-HT, GABA, DA, IL-1, BDNF and expression of 5-HT1A receptor and GABAA receptor α1-subunit in mice brain. Primary structure of peptides was identified by HPLC-ESI-QqTOF-MS/MS.
Compared with the model group, the content of 5-HT, GABA, IL-1, BDNF in mice brain of PMM peptide groups was increased to varying degrees, the content of DA was decreased, and the gene transcription and protein expression of 5-HT1A receptor and GABAA receptor α1-subunit were almost all returned to normal levels. In addition, the primary structures of most abundant nine typical peptides in PMM peptides were identified.
The results showed that PMM peptides could improve the disorder of neurotransmitter system, restore compensatory over-expression 5-HT1A receptor and GABAA receptor α1-subunit, and have a good sleep-promoting effect. The specific amino acid composition, sequence and glycosylation modification of PMM peptides may be the key reason for their activity, which lays a foundation for the subsequent development of sleep-promoting peptide products.
几个世纪以来,中华鳖被高度视为一种有同源作用的药食两用中草药。它被认为具有镇静、滋养大脑和促进睡眠的作用。然而,文献中缺少其促进睡眠活性的动物实验证据。
本研究采用 PCPP 诱导的失眠模型,探讨中华鳖酶解肽促眠的作用机制,并对其主要成分和化学结构进行分析。
采用 PCPP 诱导的失眠模型进行实验,所有动物连续 2 天腹腔注射 PCPP(350mg/kg·d)。通过测量小鼠大脑中 5-HT、GABA、DA、IL-1、BDNF 的含量以及 5-HT1A 受体和 GABAA 受体 α1 亚基的表达,评价促眠作用。采用 HPLC-ESI-QqTOF-MS/MS 鉴定肽的一级结构。
与模型组相比,PMM 肽组小鼠大脑中 5-HT、GABA、IL-1、BDNF 的含量不同程度增加,DA 的含量降低,5-HT1A 受体和 GABAA 受体 α1 亚基的基因转录和蛋白表达基本恢复正常水平。此外,还鉴定了 PMM 肽中 9 种含量丰富的典型肽的一级结构。
结果表明,PMM 肽可以改善神经递质系统紊乱,恢复代偿性过表达的 5-HT1A 受体和 GABAA 受体 α1 亚基,具有良好的促眠作用。PMM 肽的特定氨基酸组成、序列和糖基化修饰可能是其活性的关键原因,为后续开发促眠肽产品奠定了基础。