地卷柏中肽类化学成分及其神经保护活性研究

Chemical constituents of peptides from Selaginella prostrata and their neuroprotective activity

研究地卷柏Selaginella prostrata全草中的肽类化学成分及其神经保护活性。基于茚三酮试剂显色的薄层原位化学反应导向分离方法,综合运用正、反相硅胶柱色谱和半制备液相色谱等多种分离技术进行分离纯化,通过波谱数据分析和文献比对鉴定化合物的结构。共从地卷柏全草的95%乙醇提取物中分离得到15个肽类化合物,分别鉴定为pulvpeptin A(1)、nocardiotide A(2)、dianthin C(3)、longicalycinin A(4)、橙黄胡椒酰胺(5)、橙黄胡椒酰胺乙酸酯(6)、环-(L-酪氨酸-L-脯氨酸)(7)、环-(L-羟脯氨酸-L-亮氨酸)(8)、环-(L-亮氨酸-L-异亮氨酸)(9)、环-(L-脯氨酸-L-苯丙氨酸)(10)、环-(L-缬氨酸-L-脯氨酸)(11)、环-(L-脯氨酸-L-亮氨酸)(12)、环-(L-脯氨酸-L-异亮氨酸)(13)、环-(L-缬氨酸-L-苯丙氨酸)(14)、环-(L-缬氨酸-L-缬氨酸)(15)。所有化合物均为首次从地卷柏中分离获得,化合物2~4、7~15为首次从卷柏属植物中分离获得。通过建立H 2O 2诱导SH-SY5Y细胞损伤模型,采用MTT法评价分离所得化合物的神经保护活性。结果显示在50μmol/L浓度下,化合物5、6和8显示出较好的神经保护活性。本研究为地卷柏的开发利用研究提供了理论基础。

The peptide chemical constituents and neuroprotective activity in the whole herb of Selaginella prostrata were investigated.Thin-layer in situ chemical reaction oriented separation method based on ninhydrin reagent color development was used to separate and purify the compounds by positive and reversed phase silica gel column chromatography and high performance liquid chromatography.The structures of the compounds were identified by spectral data analysis and literature comparison.Fifteen peptides compounds were isolated from 95%ethanol extract of whole herb of S.prostrata.They were identified as pulvpeptin A(1),nocardiotide A(2),dianthin C(3),longicalycinin A(4),aurentiamide(5),aurantiamide acetate(6),cyclo-(L-Tyr-L-Pro)(7),cyclo-(L-Hyp-L-Leu)(8),cyclo-(L-Leu-L-Ile)(9),cyclo-(L-Pro-L-Phe)(10),cyclo-(L-Val-L-Pro)(11),cyclo-(L-Pro-L-Leu)(12),cyclo-(L-Pro-L-Ile)(13),cyclo-(L-Val-L-Phe)(14),cyclo-(L-Val-L-Val)(15).All the compounds were isolated from S.prostrata for the first time.Moreover,compounds 2-4 and 7-15 were isolated from Selaginella genus for the first time.SH-SY5Y cell damage model induced by H 2O 2 was established by cell culture in vitro.The neuroprotective activity of the isolated compounds was evaluated by MTT assay.Compounds 5,6 and 8 were showed good neuroprotective activity at the concentration of 50μmol/L.This study provided a theoretical basis for the research on the exploitation and utilization of S.prostrata.