中国中药杂志

2019, v.44(15) 3213-3220

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蛇足石杉内生真菌的分离鉴定及其代谢产物乙酰胆碱酯酶抑制活性及抗炎活性研究
Isolation and identification of endophytic fungi from Huperzia serrata and their metabolites' inhibitory activities against acetylcholinesterase and anti-inflammatory activities

齐博文;莫婷;张鑫;闫雅如;徐溪平;杨洪芸;王晓晖;李军;史社坡;刘晓;
QI Bo-wen;MO Ting;ZHANG Xin;YAN Ya-ru;XU Xi-ping;YANG Hong-yun;WANG Xiao-hui;LI Jun;SHI She-po;LIU Xiao;Modern Research Center for Traditional Chinese Medicine,School of Chinese Materia Medica,Beijing University of Chinese Medicine;Ningxia Hui Medicine Research Institute;

摘要(Abstract):

从蛇足石杉中分离鉴定出27株内生真菌,其中茎和叶中分布较多,根中分布较少;用r DNA-ITS序列分析技术对菌株进行了种属鉴定,其中1株属于担子菌门,其余26株菌分属于子囊菌门3纲5目6科9属26种,其中优势菌株为子囊菌门子囊菌纲小丛壳目小丛壳科刺盘孢属,占总菌株的48. 15%;分别采用Ellman's法和Griess法对分离得到的内生真菌发酵粗提物的乙酰胆碱酯酶抑制活性和抗炎活性进行筛选发现,4株内生真菌粗提物对乙酰胆碱酯酶具有一定的抑制作用,半数抑制浓度(IC50)为42. 5~62. 4 mg·L~(-1);部分内生真菌粗提物对LPS诱导的小鼠巨噬细胞RAW264. 7 NO的产生具有抑制作用,IC50为2. 2~51. 3 mg·L~(-1),显示其具有潜在的抗炎活性;进一步通过LCMS-IT-TOF分析对其中一株内生真菌HS21的乙酸乙酯提取物进行了化学成分分析,共鉴定17个单体化合物,包括6个聚酮类、4个二苯醚类、7个杂萜类。
A total of 27 endophytic fungal strains were isolated from Huperzia serrata,which were richly distributed in the stems and leaves while less distributed in roots. The 27 strains were identified by Internal Transcribed Spacer( ITS) r DNA molecular method and one of the strains belongs to Basidiomycota phylum,and other 26 stains belong to 26 species,9 general,6 families,5 orders,3 classes of Ascomycota Phylum. The dominant strains were Colletotrichum genus,belonging to Glomerellaceae family,Glomerellales order,Sordariomycetes class,Ascomycota Phylum,with the percentage of 48. 15%. The inhibitory activities of the crude extracts of 27 endophytic fungal strains against acetylcholinesterase( ACh E) and nitric oxide( NO) production were evaluated by Ellman's method and Griess method,respectively. Crude extracts of four fungi exhibited inhibitory activities against ACh E with an IC50 value of 42. 5-62. 4 mg·L~(-1),and some fungi's crude extracts were found to inhibit nitric oxide( NO) production in lipopolysaccharide( LPS)-activated RAW264. 7 macrophage cells with an IC50 value of 2. 2-51. 3 mg·L~(-1),which indicated that these fungi had potential anti-inflammatory activities.The chemical composition of the Et OAc extract of endophytic fungus HS21 was also analyzed by LCMS-IT-TOF. Seventeen compounds including six polyketides,four diphenyl ether derivatives and seven meroterpenoids were putatively identified.

关键词(KeyWords): 蛇足石杉;内生真菌;乙酰胆碱酯酶;抗炎活性;ITS序列分析
Huperzia serrate;endophytic fungi;acetylcholinesterase;anti-inflammatory activities;ITS sequence

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81402809);; 北京中医药大学杰出青年人才项目(2018-JYB-XJQ006);; 中国科协青年人才托举项目(CACM-2018-QNRC1-02);; 北京市自然科学基金项目(7192112)

作者(Author): 齐博文;莫婷;张鑫;闫雅如;徐溪平;杨洪芸;王晓晖;李军;史社坡;刘晓;
QI Bo-wen;MO Ting;ZHANG Xin;YAN Ya-ru;XU Xi-ping;YANG Hong-yun;WANG Xiao-hui;LI Jun;SHI She-po;LIU Xiao;Modern Research Center for Traditional Chinese Medicine,School of Chinese Materia Medica,Beijing University of Chinese Medicine;Ningxia Hui Medicine Research Institute;

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