一种羊肚菌源多肽MIP-16的分离纯化及其神经保护活性  

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

作　　者：熊川[1] 黄文丽[1] 金鑫[1] 罗强[2] 李萍[1] 张璐 李强 朱宇[1] XIONG Chuan;HUANG Wen-li;JIN Xin;LUO Qiang;LI Ping;ZHANG Lu;LI Qiang;ZHU Yu(Biotechnology and Nuclear Technology Research Institute,Sichuan Academy of Agricultural Sciences,Chengdu 610061,China;Key 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;College of Pharmacy and Biological Engineering,Chengdu University,Chengdu 610106,China)

机构地区：[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　　处：《天然产物研究与开发》2021年第9期1519-1526,共8页Natural Product Research and Development

基　　金：四川省农业科学院高新领域扩展工程(2018GXTZ-001);四川省科学技术协会创新和服务能力提升项目(2018QNRC-007)。

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1762 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。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 1762 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.

关 键 词：梯棱羊肚菌 多肽 凋亡 神经保护活性 

分 类 号：Q938.2[生物学—微生物学] Q936