Constituents of Chinese Drug Fangfeng, the Root of Saposhnikovia divaricata

从防风根的乙醇浸出物中分离鉴定出8个化合物。通过理化数据及光谱分析,分别鉴定为香柑内酯(Ⅰ),花椒毒内酯(Ⅱ),亥茅酚(Ⅲ),异紫花前胡内酯(Ⅳ),东茛菪内酯(Ⅴ),香草酸(Ⅵ),D-甘露醇(Ⅶ)和蔗糖(Ⅷ),其中Ⅳ与Ⅵ为首次自此植物中提得。

滇苦菜(Picris divaricata Vant.)对锌的吸收和富集特性

通过野外调查和营养液培养,研究Zn对滇苦菜(Picris divaricata Vant.)生长的影响及其吸收富集Zn的特性。野外调查发现,铅锌矿区土壤Zn全量范围为1724～134973mg·kg-1,平均为61495mg·kg-1。滇苦菜地上部Zn含量范围为1214～18339mg·kg-1,平均为5911mg·kg-1,且转运系数(S/R)的平均值为2.21,大于1。在营养液培养条件下,当Zn浓度达到80mg·mL-1时,滇苦菜生长开始出现明显的中毒症状,随着Zn处理的增加,植物地上部与根部的生物量呈下降趋势,Zn含量呈上升趋势,且地上部与根部在160mg·mL-1时Zn含量达最高值,分别为12472mg·kg-1和14026mg·kg-1,体内Zn富集量也达最高值1518μg.株-1,并且整个植株富集的Zn有75%～91%分布在地上部。结果表明滇苦菜具有很强的忍耐、吸收和富集Zn的能力,是我国境内发现的又一种Zn超富集植物。

防风(Saposhnikovia divaricata)组织培养中的玻璃化现象研究

以防风茎段为外植体，建立了组织培养再生体系，对防风试管苗玻璃化现象进行了研究。结果表明，与正常苗相比，玻璃化苗形态异常，组织含水量升高，叶绿素含量显著降低，酸性过氧化物同工酶活性减弱。6-BA浓度超过2．0mg／L、光照低于2000Ix、培养瓶内湿度大都极易导致防风试管苗的玻璃化，减少愈伤继代次数，增加培养基内琼脂和蔗糖浓度，可以降低玻璃化率。轻中度的玻璃化苗通过改变培养环境可以恢复正常。优化的防风再生体系为：以嫩茎段为外植体，继代3次左右的愈伤组织诱导出芽，芽继代增殖时，6-BA浓度采用1．0mg／L和0．5mg／L交替使用，培养光照3000～4000Ix。

防风（Saposchnikovia divaricata）体细胞在继代培养过程中的染色体变异

对防风（Ｓａｐｏｓｃｈｉｋｏｖｉａｄｉｖａｒｉｃａｔａ）体细胞在组织培养过程中遗传物质的稳定性进行了研究，结果发现防风体细胞在继代培养过程中染色体变异显著，有亚倍体、超倍体、单倍体、多倍体和微核细胞出现，并发生染色体的形态变异。培养半年以上亚倍体和微核细胞出现并开始增多，超倍体和多倍体细胞极少；随着培养时间的延长，亚倍体和微核细胞消失，而多倍体和超倍体细胞显著增多。对于这种规律性的变化，认为是培养基成分对不同的变异类型加以选择的结果，推测培养后期超倍体的大量出现与微核细胞的消失可能有着某种因果联系。微核可能有其它的形成途径。

New cytotoxic cerebroside from Gynura divaricata

A new cerebroside, gynuraoside （1）, was isolated from the aerial parts of Gynura divaricata DC. It was determined to be 1-0-13- o-glucopyranosyl-（2S,3 S,4R, 10E）-2- [（2＇R）-2^-hydroxyldocosanoyl-amino]- 10-octadecene- 1,3,4-triol on the basis of chemical and spectroscopic evidence. This compound showed strong cytotoxicity against L1210 leukemia cell line in vitro. 2009 Hong Tao Song. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

不同降水量对防风(Saposhnikovia divaricata)幼苗生长状态的影响

通过对不同水份条件下防风幼苗的根冠比、根粗和根重三项形态指标及幼苗中丙二醛、可溶性糖及叶绿素含量三项生理指标的测定,证明了在以甘南县为代表的黑龙江省西部半干旱气候条件下,防风幼苗在降水量超过40mm情况下可以正常生长。人工栽培时要根据实际情况处理,如苗期降水量少于40mm,幼苗会受到干旱胁迫,要依据缺水的程度补足水份或选择在雨水充足的季节之前播种。在本实验进行的气候和土壤条件下,除降水量外,根冠比、根粗、根重、丙二醛含量、可溶性糖含量和叶绿素含量也可作为判断防风幼苗能正常生长的依据。降水量低于40mm时随着干旱胁迫的加深,丙二醛和可溶性糖含量不断提高,而叶绿素含量逐渐降低。

Antifertility effect of chronically administered Tabernaemontana divaricata leaf extract on male rats

Objective:To investigate the antifertility effect of chronically administered Tabernaemontana divarkala(T.divaricata) leaf extract on male rats.Methods:The effect of 50%ethanol extract of T.divaricata leaves on reproduction was studied on male rats.The study was divided into four groups of five animals each.The first groups(1) received vehicle alone to serve as control. The second,third and fourth groups(Ⅱ,ⅡandⅣ) of animals were administered the leaf extract daily at 50 mg/kg body weight,p.o.,100 nig/kg body weight,p.o.,and 200 mg/kg body weight, p.o.,respectively,for a period of 60 days.Results:Significant decreases in the weight of testes, epididymis,seminal vesicle and ventral prostate were observed.A dose related reduction in the testicular sperm count,epididymal sperm count and motility,number of fertile male,ratio between delivered and inseminated females and numbers of pups were observed.The testis showed a clear correlation between the dose and severity of lesions of seminiferous epithelium. In general,the seminiferous tubules appear reduced in size with a frequently filled eosinophilic material.Spermatogenesis arrested at the secondary spermatocyte stage.Pachytene spermatocytes were undergoing degeneration.Disorganisation and sloughing of immature germ cell were visible. Leydinf cells were atrophied.No morphological changes were observed in Sertoli cells.Significant reduction in serum concentration of luteinizing hormone and testosterone were observed.No distinct change in serum FSH concentration was recorded.The final body weights of all groups were elevated markedly.No alterations were recorded in any hematologiocal parameters. Conclusions:It is concluded that the 50%ethanol extract of T.divaricata leaf produced dose related effect on male reproduction without altering general bodv metabolism.

Experiment on Tissue Culture Technique of Saposhnikovia divaricata(Turcz.) Schischk

The study aimed to optimize the induction and differentiation medium by exploreing different tissue culture of Saposhnikovia divaricata （Turcz.） Schischk. In tissue culture with the root, stem segments, young leaf, cotyledonary node and axillary bud of Saposhnikovia divaricata （Turcz.） Schischk as explants, a lot of plantleles were obtained and the corresponding plant regeneration-system was established. The results showed that when use MS＋1.0 mg·L^-1 6-BA＋0.2 mg·L^-1 NAA as callus induction medium, the cotyledonary node had the highest bourgeon rate, and its callus was better than any others; MS＋2 mg·L^-1 6-BA＋0.4 mg·L^-1 NAA was the best adventitious buds induction medium, and the best adventitious buds induced condition was 3% sucrose as carbon source, illumination for 12-14 h·d^-1 and pH 5.8, The best rootage medium was 1/2 MS＋0.5 mg·L^-1 NAA.

Three New Indole Alkaloids from Tabernaemontana divaricata

Three new monoterpene indole alkaloids,3a-hydroxymethyl-ibogamine(1),3a-acetatemethoxyl-ibogamine(2),16a-hy-droxyl-ibogamine(3)together with six known alkaloids were isolated from the branches and leaves of Tabernaemontana divaricata(Apocynaceae).The structures of these alkaloids were determined by spectroscopic analyses.All isolated compounds showed no significant cytotoxicity against SGC-7901 gastric cancer,HeLa,and A-549 lung cancer cell lines(IC50>20μM).

New Iboga‑Type Indole Alkaloids from Tabernaemontana divaricata

Two hitherto unknown iboga-type indole alkaloids,namely(3R)-7,19-di-epi-3-methoxytabernoxidine(1)and(3R,19R)-19-hydroxy-3-(2-oxopropyl)voacangine(2),together with eight known alkaloids(3-10),were isolated from the twigs and leaves of Tabernaemontana divaricata.Their structures were established on the basis of spectroscopic data interpretation,single crystal X-ray diffraction analysis and circular dichroism spectrum.

A New Sesquiterpene-substituted Benzoic Acid from the Brown Alga Dictyopteris divaricata

A new sesquiterpene-substituted benzoic acid has been isolated from the brown Alga Dictyopteris divaricata Okam.. Its structure was elucidated as 3-[(2-hydroxy-2,5,5,8a-tetra- methyldecahydro-1-naphthalenyl)methyl]-4-hydroxybenzoic acid, named dictyvaric acid on the basis of spectroscopic methods including IR, HRFABMS, 1D and 2D NMR techniques.

Antioxidant and Antimicrobial Activities of Larrea Divaricata Cav. Aqueous Extract on Vitamin C from Natural Orange Juice

Larrea divaricata Cav. is a South American plant widely distributed in Argentina with reported antioxidant and antim- icrobial activities. Vitamin C (Vit C) the most important water-soluble antioxidant, is degraded upon factors such as oxygen, heat, light, storage time and temperature, during cooking and in presence of microorganisms. The aim of this work was: to study the antioxidant and antimicrobial activity of an aqueous extract of L. divaricata (E) on Vit C pre- pared in water and on Vit C from natural orange juice, submitted to different store conditions such as temperatures and oxygen. The antioxidant activity was determined by DPPH method and used to obtained the relative Vit C concentra- tion;Vit C decreased its activity and concentration under temperature and air oxygen, but E allowed Vit C to recover its antioxidant activity and prevented Vit C microorganism contamination. These studies were performed in view to es- tablish whether L. divaricata can be used as a supplement on orange juices capable to prevent Vit C decay and micro- bial growth.

A NEW DOLASTANE DITERPENE PROM DICTYOTA DIVARICATA

Deacetylamijidictyol, a new dolastane diterpene was isolated from the marine brown algae Dictyota divaricata. The chemical structure was elucidated on the basis of spectral analysis.

黄东海奇异指纹蛤Acila mirabilis和指纹蛤Acila divaricata群体遗传多样性及进化研究

指纹蛤属Acila贝类是我国黄东海重要的底栖生物,研究其遗传多样性及进化对了解这一海区环境的变化及与生物的关系具有重要作用。本研究通过线粒体COI基因标记,分析了黄东海常见的两个指纹蛤属物种—奇异指纹蛤Acila mirabilis和指纹蛤Acila divaricata的分化情况,结果表明这两个种的分化形成时间分别在3.71和4.27百万年前,处于上新世时期,我们推测冰期时海平面下降引起的物种栖息地的缩减以及黄海和东海环境条件的不同是导致物种分化的重要原因。通过群体遗传多样性分析,我们发现分布于黄海的4个奇异指纹蛤群体中,3500-10群体的遗传多样性水平最高,且群体遗传多样性自冷水团中心内侧至外侧呈递减趋势,推测这可能与这一群体对冷水团有较好的适应性有关。分布于东海的3个指纹蛤群体均检测到两个单倍型类群ZA和ZB,两类群的分化时间大约在64万年前,发生于更新世中期,我们认为冰期时海平面升降引起的群体地理隔离与二次接触是导致指纹蛤两个单倍型类群形成的主要原因。

模拟降水量变化对防风(Saposhnikovia divaricata)幼苗光合特性及矿质营养积累的影响

为研究降水量变化对防风(Saposhnikovia divaricata)幼苗光合特性及矿质营养积累的影响,采用盆栽实验,以防风幼苗为材料,模拟研究了正常降水(CK,施水量为309mm,参照防风种子采集地1979-2000年6-9月的总降水量)、增加降水30%(+W)和减少降水30%(-W)对防风幼苗光合特性及矿质营养积累的影响。结果表明:与CK和+W处理相比,?W处理防风幼苗的叶片叶绿素和类胡萝卜素含量显著增加,叶片净光合速率、气孔导度和表观叶肉导度(移栽后73d、102d和128d)显著提升;脯氨酸含量(叶片、叶柄和根)、氮含量(叶片、叶柄和根)、磷含量(叶片和根)、钾含量(叶片和叶柄)、钙含量(根)和硫含量(叶片)显著增加。在本实验条件下,减少降水处理能够使防风幼苗叶片光合作用及植株矿质营养含量维持在较高水平,有利于其生长发育,有利于防风幼苗初生代谢产物的积累。

HPLC Fingerprint Combined with Chemometrics and Network Pharmacology for Q-Markers Prediction Analysis of Saposhnikovia divaricata

Saposhnikovia divaricata(SD)has high medicinal and edible value,but relatively little research has been done on its qual-ity markers(Q-markers).To further clarify the Q-markers of SD with their corresponding pharmacodynamic targets.In this experiment,14 batches of SD were identified and screened for Q-marker candidate components using a combination of HPLC fingerprint with similarity analysis,principal component analysis,hierarchical cluster analysis,and partial least squares discriminant analysis.Then,network pharmacology was used to predict Q-markers and core targets.The results showed that 5-O-methylvisammioside,cimifugin,and prim-O-glucosylcimifugin could be used as Q-markers of SD;while,MAPK1,MAPK3,PIK3CA,JUN,and MAPK8 were the core targets of SD for drug efficacy.To further evaluate the bind-ing efficiency of Q-markers,molecular docking of the main active ingredients of SD to the core targets was performed.The results showed that the compounds bind well to their targets,and binding energies were all less than-5 kcal/mol.The Q-markers obtained from the screening were closely related to the core target genes,which could achieve therapeutic effects by modulating the relevant signaling pathways.This study offers a reference for the establishment of a set of quality control evaluation system for SD potential Q-markers prediction analysis,and lays the foundation for elucidating the mechanism of actionunderlying itspharmacodynamic substance.

Transcriptome analysis of Saposhnikovia divaricata and mining of bolting and fowering genes

Objective: Early bolting of Saposhnikovia divaricata has seriously hindered its medicinal value and sustainable development of resources. The molecular mechanism of bolting and fowering of S. divaricata is still unclear and worth of research. In our study, we explored the transcriptome of the genes related to the bolting and fowering of S. divaricata.Methods: The transcriptome library was constructed, sequenced, assembled and annotated from the bolting and unbolting leaves of S. divaricata by high-throughput sequencing at the bud and fowering stage.Focus on the pathways related to bolting and fowering in plants, and exploring genes. The expression of seven candidate genes was verified by real-time fuorescence quantitative PCR(qRT-PCR).Results: Transcriptome results showed that 249 889 422 high-quality clean reads were obtained. A total of 67 866 unigenes were assembled with an average length of 948.1 bp. Trinity de Novo assembly produced 67 866 unigenes with an average length of 948.1 bp. Among 993 differentially expressed genes,484 genes were significantly up-regulated and 509 genes were down-regulated in the SdM group. A total of 79 GO terms were significantly enriched for differentially expressed genes. KEGG results showed that 11 154 unigenes were enriched in 89 pathways. And 21 candidate genes related to bolting and fowering of S. divaricata were excavated. The qRT-PCR results showed that expression trends of HDA9, PHYB, AP2,TIR1, Hsp90, CaM, and IAA7 were consistent with transcriptomic sequencing results. In addition, RNA-seq had identified 10 740 transcription factors and classified them into 58 families by their conserved domains. Further studies showed that the transcription factors regulating the fowering of S. divaricata were mainly distributed in the NAC, MYB_related, HB-other, ARF, and AP2 families.Conclusion: Based on the results of this study, it was found that the plant hormone signal transduction pathway was one of the decisive factors to control bolting and fowering. Among them, auxin related genes IAA and TIR1 are the key genes in the bolting and fowering process of S. divaricata.

防风多糖体内外免疫调节活性研究

目的采用体内外多指标免疫调节活性评价方法,研究防风多糖体内外免疫功能的调节作用。方法体外实验选用RAW 264.7小鼠巨噬细胞系,采用CCK-8法检测巨噬细胞的增殖能力,中性红染色法检测巨噬细胞的吞噬能力,Griess法测定巨噬细胞NO释放量,酶联免疫吸附法(ELISA)测定巨噬细胞肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)释放量。体内实验选用环磷酰胺诱导的免疫低下BALB/c小鼠,通过碳粒廓清实验探讨防风多糖对小鼠网状内皮系统吞噬功能,以小鼠免疫器官指数评价防风多糖对小鼠免疫器官的影响,并采用ELISA法测定小鼠体内免疫球蛋白(IgM、IgG、IgA)和补体(C3、C4)含量。结果体外实验结果表明,防风多糖能够显著提高巨噬细胞的增殖和吞噬能力,并促进巨噬细胞释放NO、TNF-α和IL-6,但对IL-1β的释放量影响不显著。体内实验结果表明,防风多糖可使免疫低下小鼠模型的吞噬能力和免疫器官指数显著提高,并使免疫低下小鼠体内的免疫球蛋白(IgM、IgG、IgA)和补体(C3、C4)的含量升高。结论防风多糖在体内外水平均具有较好的免疫调节活性,参与调节机体的特异性免疫和非特异性免疫。

防风提取物对IgE致敏肥大细胞的改善作用及机制研究

[目的]探究防风(Saposhnikovia divaricata,SD)提取物对大鼠嗜碱性白血病细胞RBL-2H3脱颗粒的影响及作用机制。[方法]采用噻唑蓝(methylthialazole tetrazolium,MTT)比色法检测,根据5、25、50、100、200、400μg·mL^(-1)SD提取物对RBL-2H3细胞活性的影响,确定后续实验浓度。以免疫球蛋白E(immunoglobulinE,IgE)诱导建立RBL-2H3细胞脱颗粒模型。设立空白对照组、模型组、低剂量SD提取物组(5μg·mL^(-1))、中剂量SD提取物组(25μg·mL^(-1))、高剂量SD提取物组(50μg·mL^(-1))和地塞米松(dexamethasone,DXMS)组(100μg·mL^(-1)),干预30 min。MTT法检测低、中、高剂量SD提取物对RBL-2H3细胞脱颗粒模型活性的影响。甲苯胺蓝染色观察脱颗粒细胞形态,计算细胞脱颗粒率。免疫荧光染色测定细胞F-肌动蛋白(F-actin)表达。酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测细胞β-氨基己糖苷酶、组胺、白细胞介素-4(interleukin-4,IL-4)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、干扰素-γ(interferon-γ,IFN-γ)水平。免疫印迹法检测细胞磷脂酰肌醇-3-羟基激酶(phosphoinositide-3 kinase,PI3K)、磷酸化-PI3K(phosphorylation-PI3K,p-PI3K)、蛋白激酶B(protein kinase B,AKT)、磷酸化-AKT(phosphorylation-AKT,p-AKT)、p38丝裂原活化蛋白激酶(p38 mitogen activited protein kinaseelisa,p38MAPK)、磷酸化-p38MAPK(phosphorylation-p38MAPK,p-p38MAPK)、核因子-κB(nuclear factor-κB,NF-κB)、磷酸化-NF-κB(phosphorylation-NF-κB,p-NF-κB)、细胞外调节激酶(extracellular regulated kinases,ERK)、磷酸化-ERK(phosphorylation-ERK,p-ERK)蛋白表达。[结果]低、中、高剂量防风提取物(5、25、50μg·mL^(-1))对RBL-2H3细胞活性无显著影响(P>0.05)。与空白对照组比较,模型组甲苯胺蓝染色细胞数量减少、形态变圆,细胞脱颗粒率显著上升,F-actin表达下降,β-氨基己糖苷酶、组胺、IL-4、IL-6、TNF-α水平升高,IFN-γ水平降低,p-PI3K/PI3K、p-AKT/AKT、p-p38MAPK/p38MAPK、p-NF-κB/NF-κB、p-ERK/ERK表达升高(P<0.01)。与模型组比较,低、中、高剂量SD提取物组和DXMS组细胞F-actin表达增加,β-氨基己糖苷酶、组胺、IL-4、IL-6、TNF-α释放显著下降(P<0.05,P<0.01),IFN-γ释放显著增加(P<0.01),p-PI3K/PI3K、p-AKT/AKT、p-p38MAPK/p38MAPK、p-NF-κB/NF-κB、p-ERK/ERK表达降低(P<0.05,P<0.01);中、高剂量SD提取物组和DXMS组细胞数量增加,形态多呈梭形,细胞脱颗粒率显著下降(P<0.01)。与低剂量SD提取物组比较,高剂量SD提取物组和DXMS组细胞脱颗粒率下降(P<0.01)、F-actin表达增加(P<0.05)、p-p38MAPK/p38MAPK表达降低(P<0.01)。[结论]SD提取物可抑制IgE致敏的RBL-2H3细胞脱颗粒,降低炎性介质水平,其作用机制可能与抑制PI3K/AKT、p38MAPK/NF-κB、ERK蛋白磷酸化有关。

防风石油醚提取物调节中性粒细胞胞外诱捕网对类风湿性关节炎大鼠的改善作用及机制

目的 探究防风石油醚提取物通过调节中性粒细胞胞外诱捕网(NETs)对类风湿性关节炎(RA)大鼠的改善作用及机制。方法 采用牛Ⅱ型胶原与弗氏完全佐剂建立大鼠RA模型。将造模成功的大鼠随机分为模型组和防风石油醚提取物低、中、高剂量组(55、110、220 mg/kg),另设不造模的正常组,每组10只。各组大鼠灌胃相应药液或等体积2%吐温-80溶液,每天1次,连续28 d。观察大鼠足趾肿胀度并进行关节炎指数(AI)评分,检测大鼠血清中白细胞介素1β(IL-1β)、IL-10、IL-17、肿瘤坏死因子α(TNF-α)、25-羟基维生素D_3[25(OH)D_3]、髓过氧化物酶(MPO)、中性粒细胞弹性蛋白酶(NE)和NETs的水平,观察大鼠踝关节病理学形态变化,检测大鼠踝关节中瓜氨酸化组蛋白H3(CitH3)以及滑膜组织中细胞色素P450 24A1(CYP24A1)、细胞色素P45027B1(CYP27B1)、维生素D受体(VDR)、肽基精氨酸脱亚胺酶4(PAD4)的表达水平。结果 与模型组比较,防风石油醚提取物中、高剂量组大鼠给药后第14天至第28天的足趾肿胀度和AI评分均显著降低(P<0.05或P<0.01);血清中IL-1β、IL-17、TNF-α、MPO、NE和NETs水平均显著降低,IL-10、25(OH)D_3水平均显著升高(P<0.05或P<0.01);踝关节间隙变宽,结构明显改善;踝关节中CitH3以及滑膜组织中CYP24A1、PAD4表达水平均显著降低,滑膜组织中CYP27B1、VDR表达水平均显著升高(P<0.05或P<0.01)。结论 防风石油醚提取物可能通过调控维生素D系统,抑制NETs生成,进而改善RA症状。