Isolation and identification of endophytic fungi from Huperzia serrata and their metabolites' inhibitory activities against acetylcholinesterase and anti-inflammatory activities / 中国中药杂志

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.

Phenolic Glycosides from the Aerial Part of Urena lobata / 中国药学杂志

OBJECTIVE: To study the phenolic glycosides of Urena lobata. METHODS: Compounds were isolated and purified using various column chromatographies such as D101 macroporous adsorption resin, silica gel, Sephadex LH-20, and preparation HPLC. Their structures were established using extensive spectroscopic techniques such as MS and NMR. The anti-inflammatory activities of all compounds were evaluated by using LPS-stimulated RAW264.7 cells. RESULTS: Twelve phenolic glycosides were obtained from the n-BuOH extract of U. lobata including benzyl-7-O-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranoside (1), phenylethyl-8-O-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranoside (2), phenylethyl-8-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside (3), eugenyl-1-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside (4), 6-methoxyeugenyl-1-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside (5), 3,4,5-trimethoxybenzene-1-O-α-L-rhamnosyl-(1→6)-β-D-glucopyranoside (6), 4-O-(glycer-2-yl)-dihydroconiferylalcohol-1′-O-β-D-mannopyranoside (7), benzyl-7-O-β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside (8), 3,5-dimethoxy-4-hydroxyphenylethyl-8-O-β-D-glucopyranoside (9), 3,4,5-trimethoxybenzene-1-O-β-D-glucopyranoside (10), phenylethyl-8-O-β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside(11),and 3,4,5-trimethoxybenzene-1-O-β-D-apiofuranosyl-(1→6)-β-D-glucopyranoside (12). CONCLUSION: Compounds 2-8 and 10-12 are obtained from the family Malvaceae for the first time, and compounds 3, 8, 9 and 11 show moderate inhibition of nitric oxide production in LPS-stimulated RAW264.7 cells, with IC50values of(40.5±4.9 ), (31.9±4.6),( 37.7±3.3 ), and (36.1±4.6 )μmol•L-1, respectively.

Cloning and expression analysis of S-adenosylmethionine synthetase gene from Aquilaria sinensis / 药学学报

S-adenosylmethionine synthetase, a key enzyme in plant metabolism, plays an essential role in the plant defence system. In present study, a full length cDNA sequence of AsSAMS1 gene was cloned by RACE and reverse transcription PCR from Aquilaria sinensis calli. Meanwhile, the bioinformatics, prokaryotic expression, tissue-specific expression analysis, and expression analysis under different abiotic stresses and hormone treatments were performed. The open reading frame (ORF) of AsSAMS1 gene was 1 183 bp, encoding a protein of 393 amino acids with a calculated molecular mass (MW) of 43.13 kDa. Bioinformatic analysis indicated that AsSAMS1 contained 3 SAMS characteristic sequences. The phylogenetic analysis indicated that AsSAMS1 protein had the highest level of homology with SAMS protein from Glycine soja. The recombinant AsSAMS1 protein was successfully expressed in Escherichia coli BL21 (DE3) cells using the prokaryotic expression vector pET28a-AsSAMS1 and the recombinant AsSAMS1 was purified by Ni2+ affinity chromatography. Expression analysis results in different tissues indicated that AsSAMS1 was primarily observed in stems, and then stem tips and leaves, following by roots. The transcript level of AsSAMS1 and the content of S-adenosylmethionine (SAM) were induced by various abiotic stresses including salt, drought, cold, and heavy metal stress. Furthermore, AsSAMS1 expression level was enhanced upon methyl jasmonate (MeJA), salicylic acid (SA), gibberellin (GA3), and abscisic acid (ABA) treatment. These results provided valuable insights for further study on the role of SAMS in the mechanism of agarwood formation and plant resistance.