山核桃水溶性成分中神经营养活性因子的分离与鉴定.PDFVIP

中国农学通报 2018,34(12): 159-164 Chinese Agricultural Science Bulletin 山核桃水溶性成分中神经营养活性因子的分离与鉴定 吴建峰，高 飞，谢 倩，黄坚钦，钱永常 （浙江农林大学林业与生物技术学院，浙江临安311300） 摘 要：本研究旨在揭示山核桃水溶性成分的神经营养活性及活性因子。利用脱脂山核桃水溶性提取 物处理SH-SY5Y 细胞，用倒置相差显微镜观察细胞突触生长，用qPCR 检测相关基因表达，使用HPLC 及LC-MS 对山核桃水溶性成分神经营养活性因子进行分离鉴定。结果显示，山核桃的水溶性成分能够 显著促进SH-SY5Y 细胞的突触生长，并可以显著上调SH-SY5Y 细胞NF 160基因的表达。HPLC 及LC- MS 检测结果发现其水溶性成分主要组成为胞苷、尿苷、鸟苷、腺嘌呤、腺苷等。由此可知，山核桃水溶 性成分通过上调NF 160基因表达具有神经营养活性。 关键词：山核桃；水溶性成分；SH-SY5Y 细胞；HPLC ；LC-MS ；神经营养活性 中图分类号：S789.7 文献标志码：A 论文编号：cas IsolationandIdentificationofNeurotrophicFactorsfrom Water-solubleConstituentsofCaryacathayensisSarg. WuJianfeng,GaoFei,XieQian,HuangJianqin,QianYongchang (CollegeofForestryandBiotechnology,ZhejiangUniversityofAgricultureandForestry,Lin’anZhejiang311300) Abstract: The aims are to clarify the neurotrophic activity and neurotrophic factors of water-soluble constituentsfromhickory.TheSH-SY5Ycellwastreatedwithanon-fathickorywater-solubleextract.The growthofneuriteswasdetectedbyinvertedphasecontrastmicroscope.Theexpressionofrelatedgeneswas measuredbyqPCR.Theisolationandidentification ofneurotrophicfactorswasconductedbyHPLCandLC- MS.Theresultsshowedthatthewater-solubleextractofhickorycouldpromotethegrowthofSH-SY5Y neuritessignificantly,aswellasup-regulatethegeneexpressionofNF160.TheresultsofHPLCandLC-MS showedthatthemainconstituentsofwater-solublewerecytoside,urea,guanosine,adenineandadenosine.Itis concludedthatthewater-solubleconstituentsofhickoryhasneurotrophicactivitybyup-regulatingthegene expressionofNF160. Keywords:hickory,water-solubleconstituents,SH-SY5Ycells,HPLC;LC-MS,neurotrophicactivity