天然产物研究与开发 ›› 2021, Vol. 33 ›› Issue (9): 1519-1526.doi: 10.16333/j.1001-6880.2021.9.010

• 研究简报 • 上一篇    下一篇

一种羊肚菌源多肽MIP-16的分离纯化及其神经保护活性

熊川1，黄文丽1，金鑫1，罗强2，李萍1，张璐1，李强3，朱宇1*   

  1. 1四川省农业科学院生物技术核技术研究所，成都 610061；2重庆医科大学附属第二医院 感染病分子重点实验室肝炎研究所，重庆 400010；3成都大学药学与生物工程学院，成都 610106

  • 出版日期:2021-09-28 发布日期:2021-09-28
  • 基金资助:
    四川省农业科学院高新领域扩展工程（2018GXTZ-0 01）；四川省科学技术协会创新和服务能力提升项目（2018QNRC-007）

Isolation and purification of a novel peptide from the fruiting bodies of Morchella importuna and its neuroprotective effect

XIONG Chuan1,HUANG Wen-li1,JIN Xin1,LUO Qiang2,LI Ping1,ZHANG Lu1,LI Qiang3,ZHU Yu1*   

  1. 1Biotechnology and Nuclear Technology Research Institute,Sichuan Academy of Agricultural Sciences,Chengdu 610061,China; 2Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education),Institute for Viral Hepatitis,Department of Infectious Diseases,the Second Affiliated Hospital,Chongqing Medical University,Chongqing 400010,China; 3College of Pharmacy and Biological Engineering,Chengdu University,Chengdu 610106,China

  • Online:2021-09-28 Published:2021-09-28

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摘要：

分离纯化人工栽培的梯棱羊肚菌子实体多肽（MIP-16），对其结构和神经保护活性进行研究。采用磷酸盐缓冲液提取，分子排阻色谱分离，反相高效液相色谱纯化获得梯棱羊肚菌子实体多肽，通过液相色谱-质谱连用技术完成氨基酸序列鉴定。构建6-羟基多巴胺处理后引起凋亡的PC12细胞模型，验证MIP-16对PC12细胞的保护作用。结果显示：MIP-16由16个氨基酸组成，相对分子质量为1 762 Da，氨基酸组成序列为Thr-Ile-Thr-Leu-Glu-Val-Glu-Ser-Ser-Asn-Ile-Thr-Asn-Asp-Val-Lys。细胞实验发现，MIP-16能够抑制PC12细胞丙二醛（MDA）产生，降低活性氧（ROS）水平，调节Bcl-2和Bax的比例，并降低Caspase蛋白的表达。MIP-16通过细胞线粒体途径抑制PC12细胞凋亡，具有神经保护活性，可作为一种辅助治疗帕金森综合症的药物开发。

关键词: 梯棱羊肚菌, 多肽, 凋亡, 神经保护活性

Abstract:

In this research,we isolated and purified a novel peptide (MIP-16) from the fruiting bodies of Morchella importuna M.Kuo.MIP-16 was extracted with phosphate buffer,and purified by molecular exclusion chromatography followed by reversed-phase high performance liquid chromatography (RP-HPLC).The amino acid sequence of MIP-16 was determined by LC-MS-MS as Thr-Ile-Thr-Leu-Glu-Val-Glu-Ser-Ser-Asn-Ile-Thr-Asn-Asp-Val-Lys with molecular mass of 1 762 Da.6-hydroxydopamine-induced apoptotic PC12 cell model was used to verify the protective effect of MIP-16 on PC12 cells from apoptosis.The results showed that treatment with MIP-16 significantly inhibits the production of malondialdehyde (MDA),reduces the level of reactive oxygen species (ROS),regulates the ratio of Bcl-2 and Bax,and reduces the activity of Caspase in PC12 cells;indicating its neuroprotective activity through a mitochondrial pathway.These results suggest a potential application of MIP-16 as an adjuvant therapy for the treatment of Parkinson's syndrome.

Key words: Morchella importuna, peptide, apoptosis, neuroprotective activity

中图分类号:  Q938.2 Q936

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