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

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.