Huperzine H, a New Lycopodium Alkaloid from Huperzia serrata

A new lycopodium alkaloid with a novel skeleton was isolated from Huperzia serrata.Its structure was determined on the basis of spectral evidences.

Lycodine-Type Lycopodium Alkaloids from the Whole Plants of Huperzia serrata

t Three new lycodine-type Lycopodium alkaloids,namely 1-methyllycodine(1),8a-hydroxy-15,16-dehydro-desN-methyl-a-obscurine(2),N-methyl-16-hydroxyhuperzine B(3),and one new natural lycodine-type Lycopodium alkaloid,N-methylhuperzine A(4),along with 11 known analogues(5–15),were isolated from the whole plants of club moss Huperzia serrata.The structures of 1–4 were elucidated on the basis of NMR spectroscopic and mass spectrometry data.Among them,compound 1 was the first lycodine-type alkaloid possessing a methyl group at C-1.In addition,the structure of 5 was confirmed by the single-crystal X-ray crystallography data and its^(13)C NMR was reported for the first time in current study.Compounds 1–5 were tested their BACE1 inhibitory activity.

Bioinformatic Analysis of ARF7 in Huperzia serrata

[Objectives]This paper aims to study the function of the ARF7 gene family of Huperzia serrata.[Methods]Based on the full-length transcriptome sequencing results of H.serrata,the unigenes of 17 H.serrata ARF7 candidate genes were analyzed using biogenetics technology.[Results]The molecular weight of the ARF7 proteins ranges from 11.28 to 608.54 kDa.HsARF7-4,HsARF7-5,HsARF7-8,HsARF7-9,HsARF7-10,HsARF7-13,HsARF7-16 and HsARF7-17 are basic proteins;all the proteins are unstable;except that HsARF7-6 and HsARF7-15 are hydrophilic,the rest are lipophilic proteins;there are 197 types of cis-acting elements,and 17 of the proteins contain as more as 20 acting elements such as GTGANTG10,CBFHV,GT1CONSENSUS,NODCON2GM,CAATBOX1,MYBCORE,GATABOX,WRKY71OS and RAV1AAT;HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-6,HsARF7-8,HsARF7-10,HsARF7-12,HsARF7-14 and HsARF7-16 all have the 6 types of motif predicted;the proteins have domains such as PABP-1234 super family,RRM_SF super famliy,PRK10263 super family,AUX_IAA super family and MFS super family;the proteins all can cross cell membrane and have no signal peptide,and they are all located in the nucleus.On the level of secondary structure,the proportion ofβ-sheet is the smallest.In HsARF7-7 and HsARF7-9,α-helix has the largest proportion,and in the remaining proteins,random coil has the largest proportion;and tertiary structure prediction found that HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-14 and HsARF7-17 have a tertiary structure of polyadenylate-binding protein,and HsARF7-7 has a tertiary structure of auxin-induced protein iaa4.[Conclusions]The above analysis results can provide a certain theoretical basis for further research on the biological function and regulatory mechanism of the ARF gene of H.serrata in the future,and provide a reference for the study of the growth and development of H.serrata.

Huperzine V,A New Lycopodium Alkaloid from Huperzia serrata

Huperzine V, a new Lycopodium alkaloid, was isolated from the whole plant of Huperzia serrata, and the absolute stereochemistry was determined by X-ray crystallographic analysis.

Huperzine W,a Novel 14 Carbons Lycopodium Alkaloid from Huperzia serrata

Huperzine W, a novel 14 carbons Lycopodium alkaloid, was isolated from the whole plant of Huperzia serrata, and its stucture was determined by spectroscopic analysis.

Cloning and analysis of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase genes HsHDR1 and HsHDR2 in Huperzia serrate

We cloned and analyzed the two genes of the 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase(HDR) gene family from Huperzia serrate.The two transcripts coding HDR,named Hs HDR1 and Hs HDR2,were discovered in the transcriptome dataset of H.serrate and were cloned by reverse transcription-polymerase chain reaction(RT-PCR).The physicochemical properties,protein domains,protein secondary structure,and 3D structure of the putative Hs HDR1 and Hs HDR2 proteins were analyzed.The full-length c DNA of the Hs HDR1 gene contained 1431 bp encoding a putative protein with 476 amino acids,whereas the Hs HDR2 gene contained 1428 bp encoding a putative protein of 475 amino acids.These two proteins contained the conserved domain of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase(PF02401),but without the transmembrane region and signal peptide.The most abundant expression of Hs HDR1 and Hs HDR2 was detected in H.serrate roots,followed by the stems and leaves.Our results provide a foundation for exploring the function of Hs HDR1 and Hs HDR2 in terpenoid and sterol biosynthesis in Huperziaceae plants.

Comprehensive relative quantitative metabolomics analysis of lycopodium alkaloids in different tissues of Huperzia serrata

Qian ceng Ta,the whole plant of Huperzia serrata,is an important landscape and medicinal herbs and contains abundant bioactive lycopodium alkaloids.Although the structures of more than 100 lycopodium alkaloids in Huperzia serrata have been isolated and identified,the content and distribution of these alkaloids in different tissues are still unclear.In current study,an ultra-performance liquid chromatography-mass spectrometry based comprehensive metabolomics strategy was developed,including the extraction,separation,identification,and statistical analysis.The results showed that different types lycopodium alkaloids could be separated at different time-windows,which was helpful for further metabolite identification.Peak4388 and peak3954 were metabolite biomarkers for the different tissues according to the principle component analysis and partial least squares-discriminant analysis model.A computational tool based in-house database was also built up and used for putative identification.Of the 2354 true peaks after four-step filtration,118 peaks were putatively identified as lycopodium alkaloids by using in-house database,and four of which was identified by authentic standards.Alternatively,another computational software was used to predict the fragmentation pattern,to dereplicate the structure of identified peaks,and identified the peak3585 to N-methylhuperzine A.The integration of both computational tools could be used for more metabolites identification.

A new alkaloid and arbutin from the whole plant of Huperzia serrata

A new Lycopodium alkaloid together with a phenolic glyco-side-arbutin, were isolated from the whole plant of Huperzia serrata. The new compound was assigned the trivial name of huperzine I. The structures of these two compounds were determined on the basis of spectral evidence.

Triterpenoid Constituents of Huperzia miyoshiana

Thirteen triterpenoids, including three new ones, miyoshianols A (1), B (2) and C (3), were isolated from Huperzia miyoshiana . The structures of these new compounds were established as 3 O dihydroferuloyltohogenol (1) , 16 oxo 3 β ,21 β dihydroxy serrat 14 en 24 ferulate (2) and 16 oxo 3 α ,21 β dihydroxy serrat 14 en 24 ferulate (3), respectively, on the basis of their spectroscopic analysis.

蛇足石杉仿野生栽培关键技术分析

为保护蛇足石杉野生资源,促进可持续利用,针对蛇足石杉仿野生栽培相关的适宜温湿度、光照强度、肥料效应和不同居群在当地的生长速度等部分关键技术环节展开初步研究,为后续蛇足石杉仿野生栽培体系的创建提供参考。

共生培养法促进石杉碱甲增量富集的研究

为了促进蛇足石杉中石杉碱甲增量富集,本文利用内生真菌与蛇足石杉的共生进化关系,以共培养体系(co-culture system,CCS)为基础,提出了增加石杉碱甲产率的新方法。以蛇足石杉植株存活率为指标,确定共培养体系最佳培养条件;以细胞存活率、愈伤组织诱导率和增殖率为指标,确定最佳培养基条件;以石杉碱甲和麦角甾醇为指标性成分,控制植物细胞和内生真菌比例关系,维持培养体系内植物细胞和内生真菌之间的生长平衡,获得最佳的继代时间。结果表明:CCS的最佳培养条件为光照强度100 Lux,湿度95%,MS液体培养基,蔗糖20.0 g/L,2,4-二氯苯氧基乙酸5.0 mg/L和激动素1.0 mg/L;4 d为最佳继代时间,其石杉碱甲的浓度最高,与1 d时相比增加了49.34%,并且石杉碱甲与麦角甾醇的浓度比值控制在1.5~2.2之间时,内生真菌生长对石杉碱甲的富集有良性促进作用。该研究为高效、低成本增加石杉碱甲产量提供新的探索路径,亦可以有效解决药源不足问题,具有很好的应用前景。

千层塔中脂肪族类化学成分的抗肿瘤活性研究

千层塔中脂肪族类化学成分及其抗肿瘤活性研究。在肿瘤抑制活性引导下,采用AB-8大孔树脂、氧化铝、硅胶、Sephadex LH 20和半制备液相等方法,对千层塔甲醇提取物进行较系统分离纯化,并结合现代波谱手段对分离到的化合物进行结构鉴定。共分离鉴定了9个单体化合物,分别为pentyl-4-oxoicosa-2(Z),6(Z)-dienoate(1)、2-(2-ethoxy-1-hydroxy-2-oxoethyl)-2-hydroxy-4-methylpentanoic acid(2)、ethyl (9Z,12Z)-octadeca-9,12-dienoate(3)、(Z)-nonadec-10-enal(4)、油酸酰胺(5)、反式-9-十八碳烯酸(6)、bacillamidin A(7)、(3Z,6Z,9Z)-3,6,9-二十二碳三烯(8)、oplopandiol(9)。其中,化合物1和化合物2为新化合物,化合物3~9系首次从千层塔中分离。肿瘤抑制活性结果显示,化合物1和3对HepG2细胞具有抑制作用,其IC50分别为41.12、80.54μmol/L。

蛇足石杉中石杉碱甲的酶法提取研究

[目的]筛选蛇足石杉中石杉碱甲的酶法提取的最佳提取条件。[方法]采用纤维素酶酶解细胞壁,从而溶出更多的蛇足石杉细胞中的物质,并设计正交试验选出酶法提取的最佳提取条件。[结果]蛇足石杉中石杉碱甲的酶法提取的最佳工艺条件为纤维素酶用量0.125 g,酶解pH4.5,酶解温度60℃,酶解时间2.5 h;在此条件下,蛇足石杉中石杉碱甲的提取率为0.589‰;各因素中温度对纤维素酶的酶解效果影响最大。与酸提法相比,酶法提取的石杉碱甲的提取率提高了40.3%。[结论]该方法简便快捷、提取率高,可用来提取蛇足石杉中石杉碱甲。

蛇足石杉内生真菌清除DPPH·与抑制乙酰胆碱酯酶活性研究

为了从蛇足石杉[Huperzia serrata（Thunb.）Trev.]中分离出能产生清除DPPH·和抑制乙酰胆碱酯酶活性物质的内生真菌,采用平板分离蛇足石杉内生真菌,摇瓶发酵并测定其发酵液和菌丝体提取物清除DPPH自由基、抑制乙酰胆碱酯酶活性的能力,形态学和ITS-rDNA序列分析法鉴定获得的部分内生真菌。结果表明,共分离得到94株内生真菌,有31株内生真菌发酵产物显示出乙酰胆碱酯酶抑制活性,其中18株具有显著的抑制活性,其抑制率范围在50.3%～82.78%,分别分离自蛇足石杉茎部（9株）、叶部（8株）和根（1株）。有19株内生真菌发酵产物有清除DPPH·活性,清除率在30%以上。有5株内生真菌的发酵物兼有乙酰胆碱酯酶抑制活性和清除DPPH·活性,以ITS-rDNA序列比对法对5株菌进行了鉴定,其分别属于Arthrinium sp.、Podospora sp.、Trichoderma sp.、Colletotrichum sp.、Colletotrichum sp。表明中药蛇足石杉植株内存在能产生抑制乙酰胆碱酯酶和抗DPPH自由基活性物质的内生真菌。

蛇足石杉体内抑制乙酰胆碱酯酶活性物质的内生真菌的分离

[目的]从蛇足石杉体内分离筛选出具有体外抑制乙酰胆碱酯酶活性的内生真菌。[方法]取野外采集的蛇足石杉的叶、茎和根进行表面消毒后接种于PDA培养基,分离出内生真菌后进行摇瓶发酵,超声提取其发酵产物,以DTNB法检测其发酵产物在体外抑制乙酰胆碱酯酶的活性。[结果]从根、茎、叶中分离得到194株内生真菌,在马铃薯葡萄糖液体培养基(PDB)中发酵后,对内生真菌发酵提取液进行体外AChE抑制活性检测;共有31株内生真菌显示出体外AChE抑制活性,抑制活性较强的有13株,抑制率在50.2%～80.5%。其中从蛇足石杉茎部位分离得到9株,叶部位分离得到4株。[结论]从蛇足石杉体内可分离出内生真菌,该内生真菌的发酵产物在体外有较强的抑制乙酰胆碱酯酶活性,这可能是寻找乙酰胆碱酯酶抑制药物的有效途径之一。

千层塔基原及药效文献研究

千层塔为民间常用中草药,具有清热解毒、燥湿敛疮、止血定痛等功效,现代用于治疗阿尔茨海默病,具有极大的药用价值。通过本草考证和梳理现代研究文献,对千层塔的基原、名称、采收加工、资源利用、毒性、功效主治、民族用药及其有效成分石杉碱甲,治疗阿尔茨海默病的应用,进行系统考证与综述,为进一步开发应用千层塔提供参考依据。

长柄石杉中石杉碱甲的含量测定

[目的]建立HPLC法测定长柄石杉中石杉碱甲的含量。[方法]色谱条件:色谱柱为ZORBAX Extend-C18(150 mm×4.6 mm,4.5μm),流动相为乙腈-浓度0.02 mol/L磷酸二氢钾(8∶92,V/V),检测波长为310 nm,柱温为25℃,流速为1 ml/min,进样量为10μl。[结果]石杉碱甲在0.046～0.460μg/ml浓度范围内与峰面积呈良好线性关系(R=0.999 9),平均回收率为97.45%,RSD为2.70%,平均含量为0.025%。[结论]该方法简单、易行,可用于长柄石杉中石杉碱甲的含量测定。

陕西石杉属植物2种新记录

本文报道了陕西石杉属植物省级新记录种2种,分别为金发石杉[Huperzia quasipolytrichoides(Hayata)Ching]和峨眉石杉[Huperzia emeiensis(Ching et H.S.Kung)Ching et H.S.Kung],凭证标本保存于陕西长青国家级自然保护区管理局标本室。

HPLC法测定蛇足石杉中石杉碱甲含量

目的 考察蛇足石杉不同产地、不同部位中石杉碱甲 (Hup A)和总碱含量 ;比较不同提取方法对石杉碱甲含量的影响。方法 采用 RP- HPL C法测定 Hup A的含量。结果 Hup A在根、茎、叶中的含量为茎、叶 >根 ;贵州、广东和安徽产地的 Hup A含量分别为 0 .0 18% ,0 .0 2 1% ,0 .0 2 0 % ,提取方法之间差异不显著。结论 Hup A含量在蛇足石杉中的分布为地上部分大于地下部分。不同产地 Hup A含量有一定差异。

草药蛇足石杉的研究进展

对蛇足石杉的药理作用及其临床应用方面的研究作较全面综述 。