Four new lanostane triterpenoids featuring extendedπ-conjugated systems from the stems of Kadsura coccinea

Four new 14(13→12)-abeolanostane triterpenoids featuring extendedπ-conjugated systems,kadcoccitanes E-H(1-4),were obtained from the stems of Kadsura coccinea through using a HPLC−UV-guided approach.Their structural and configurational determination was accomplished through extensive spectroscopic analysis coupled with quantum chemical calculations.Kadcoccitanes E-H were tested for their cytotoxic activities against five human tumor cell lines(HL-60,A-549,SMMC-7721,MDA-MB-231,SW-480)but none of them exhibited activities at the concentration 40μM.

Study on Effect of Drought Stress with PEG-6000 on Physiological and Biochemical Indicators in Kadsura coccinea

[Objective] The aim was to research drought resistance and adaptation mechanisms of Kaclsura coccinea under different drought stresses. [Method] The seedlings of Kadsura coccinea growing one year were taken as materials in order to investigate and analyze leaf SPAD value, relative water content （RWC）, malonaldehyde （MAD）, cell membrane permeability （RP）, proline （PRO） and superoxide dismutase activity （SOD）. [Result] As drought stress improved, leaf SPAD value and RWC were both diminishing and RWC fell remarkably; MAD, Pro, RP and SOD activity were increasing; MAD and RP grew significantly. [Conclusion] Kadsura coccinea is much sensitive to drought stress and will be affected if water loss is too much high.

A new triterpenoid from Kadsura coccinea

A new triterpenoid,possessing a lanost-9（11）-3-one skeleton,was isolated from the roots of Kadsura coccinea,based on the spectral analysis,its structure was elucidated as 3-hydroxy-12-acetoxycoccinic acid.

Four 14(13→12)‑Abeolanostane Triterpenoids with 6/6/5/6‑Fused Ring System from the Roots of Kadsura coccinea

Four new rearranged 6/6/5/6-fused lanostane-type triterpenoids,kadcoccitanes A-D(1-4),were isolated from the roots of Kadsura coccinea,and their structures were mainly elucidated by comprehensive analysis of their spectroscopic data.Additionally,the structure of 1 was ambiguously verifed by single-crystal X-ray difraction,while the structure of 2,which features a novel 8,16-epoxy motif,was validated by quantum chemical calculation of NMR parameters and ECD spectrum.Moreover,1 and 4 were found to exhibited anticoagulant activity,while 3 and 4 were found to possess anti-platelet aggregation activity.

Analysis of Growth and Quality of Sun-cured Tobacco Kadsura Coccinea In Guangchang County

[Objective] The aim was to study the growth and quality characteristics of sun-cured tobacco Kadsura Coccinea in Guangchang County and give full play to its advantages. [Method] The research took tobacco varieties including Tiechi in Yiqian, Mushao in Yangxi Tiechi in Chishui, and Liuye in Toupo as test materials to explore development process, quality characteristics and economic traits of different varieties. [Result] The variety of Tiechi in Yiqian had medium growht, moderate growth periods, and the best economic benefits. Total content of aroma matter was the highest in the varieties of Tiechi in Yiqian. Also it had more content of products of browning reaction and products degraded by cembranoids. Besides, volume and quality of aroma were higher, as well as the score of sensory quality. [Conclusion]Comprehensive comparisons suggested that the variety of Tiechi in Yiqian was superior to others at tobacco-growing, economic benefits and quality.

Experimental Study on Anti-adjuvant Arthritis in Rats of Kadsura coccinea

[Objectives] To investigate the therapeutic effect of Kadsura coccinea on adjuvant arthritis( AA) rats and its mechanism from cytokine levels. [Methods] The complete Freund's adjuvant( CFA)-induced AA model of rats was used. The therapeutic effect of K. coccinea on AA rats was evaluated by the articular swelling,arthritis score,immune organ index and hematoxylin-eosin( HE) staining. The content of inflammatory factors TNF-α,IL-6,and IL-1β in the serum of AA rats was detected by enzyme-linked immunosorbent assay( ELISA) respectively. [Results]Compared with the model group,the total extract of K. coccinea( KCTE) could significantly inhibit the primary and secondary articular swelling of AA rats,lower the arthritis score and immune organ index,and improve the pathological state of ankles of AA rats. In the groups with a high and middle dosage of KCTE( KCTE-H and KCTE-M),KCTE could significantly reduce the content of TNF-α and IL-6 in the serum of rats,exhibiting a certain dose-effect relationship. [Conclusions]KCTE had significant therapeutic effect on the AA of rats,and the mechanism might be associated with the modulation of immune function and the decrease in the secretion of inflammatory cytokines TNF-α and IL-6.

Differential metabolome landscape of Kadsura coccinea fruit tissues and potential valorization of the peel and seed tissues

Kadsura coccinea(Lem.)is a woody wine plant with a peculiar fruit enriched in important health-promoting compounds.The non-editable part of the fruit,i.e.,the seed and peel,represents more than 60%of the fruit and is considered a biowaste.This significantly restricts the development of the K.coccinea fruit industry.Clarifying the metabolic components of the different fruit parts can help to improve the utilization rate and valorization of K.coccinea.Herein,we evaluated K.coccinea fruit peel,pulp,and seed using widely-targeted metabolomics and quantified a set of 736 bioactive compounds from 11 major metabolite classes.The most prominent metabolite classes included lipids,amino acids,flavonoids,and lignans.Furthermore,our results emphasized a significant accumulation of flavonoids in pulp tissues,while alkaloids and lignans were abundant in peel and seed tissues,respectively.A total of 183 metabolites were differentially accumulated among the three tissues.Procyanidin C2,rutinoside,2-hydroxyoleanolic acid,5-hydroxymethyluracil,nootkatol,isoquercitrin,isohyperoside,quercetin-7-O-glucoside,hyperin,and rutin showed elevated accumulation in the peel.In the seed,kadsuralignan G,kadcoccilactone A,kadsuralignan H,lysoPE 20:5,iso-schisandrin ethyl alcohol,and kadangustin were significantly enriched.Our results highlight the diverse metabolome composition of K.coccinea fruit parts,which can be further exploited for its valorization in various industries.

Diurnal Variation of Photosynthetic Physiological Characteristics of Kadsura coccinea(Lem.)A.C.Smith

[Objectives]This study was conducted to investigate the diurnal variations of photosynthetic and physiological characteristics in 3-year-old Kadsura coccinea(Lem.)A.C.Smith plants.[Methods]A Li-6400 portable photosynthetic system was used to study the diurnal variations of the photosynthetic physiological characteristics of K.coccinea.[Results]The diurnal variation of the net photosynthetic rate(Pn)of K.coccinea leaves showed a"double peak"curve,and there was a lunch break at noon.The maximum value of Pn[4.51 mmol/(m^(2)·s)]appeared at 10:00,and the daily average Pn was 3.98 mol/(m^(2)·s);and the stepwise regression analysis showed that photosynthetically active radiation(PAR)and stomatal conductance(Gs)were the main factors affecting Pn.The partial correlation analysis and path analysis showed that the order of effects on Pn was Gs>PAR.[Conclusions]This study provides a reference for further research on the development and utilization of K.coccinea.

Technical Regulations for Standardized Planting of Rare and Multi-purpose Wild Fruit Kadsura coccinea

The cultivation technology were standardized for improving the yield and quality,and the commodity value and the competitiveness of fruit market,and solving the bottleneck problem—fruit quality and yield that affect the industrial development of rare and multi-purpose wild fruit Kadsura coccinea(Lem.)A.C.Smith.Through the investigation of K.coccinea production area and planting area,collection and analysis of fruit samples,it was found that in most planting area,the varieties of K.coccinea were mixed,with great differences in fruit quality and yield,which makes it impossible to realize large-scale planting.In view of these problems,a series of studies have been carried out and the technical regulations for standardized planting of rare and multi-purpose wild fruit K.coccinea were formulated.

A lignin glycoside and a nortriterpenoid from Kadsura coccinea

AIM: To study the chemical constituents of the roots and stem bark of Kadsura coccinea. METHOD: Compounds were isolated by column chromatography on silica gel and Sephadex LH-20, and finally purified by prep-HPLC. Their structures were elucidated by extensive spectroscopic methods, including 1D- and 2D-NMR, and HR-ESI-MS. RESULTS: Two compounds were determined as(7′S,8′S,8R)-(8β,8′α)- dimethyl-4,4′-dihydroxy-5,3′-dimethoxy-5′-cyclolignan glucoside(1) and micrandiactone H(2), respectively. CONCLUSION: Compunds 1 and 2 are new and neither showed inhibitory effects on nitric oxide(NO) production in lipopolysaccharide-induced RAW264.7 macrophages.

Kadsura coccinea:A rich source of structurally diverse and biologically important compounds

Kadsura coccinea belongs to medicinally important genus Kadsura from the Schisandraceae family.It has been used in traditional Chinese medicine(TCM)for the treatment of rheumatoid arthritis and gastroenteric disorders.The initial phytochemical work focused on the identification of some structurally novel and diverse natural products,which turned the attention of many researchers towards this plant.Thus far,202 compounds have been reported in this plant.Lignans and terpenoids were found as the main chemical constituents of this plant.Some of the triterpenoids and sesquiterpenoids with novel structures are of particular interest for natural product researchers.The isolated compounds of this plant have shown different bioactivities including anti-tumor,anti-HIV,anti-inflammatory,nitric oxide(NO)production inhibitory and other pharmacological effects.This review systematically summarizes all the phytochemical and pharmacological work done so far on K.coccinea,and can be used as a reference for future research on this plant.

A high-performance liquid chromatography with circular dichroism detector for determination of stereochemistry of 6, 9-oxygen bridge dibenzocyclooctadiene lignans from kadsura coccinea

The stereochemistry of two 6, 9-oxygen bridge dibenzocyclooctadiene lignans from Kadsura coccinea, are difficult to separate and very unstable. The present study was designed to develop a high-performance liquid chromatography using circular dichroism detection for the analysis of the stereochemistry. A new 6, 9-oxygen bridge dibenzocyclooctadiene lignans named Kadsulignan Q was firstly found with an S-biphenyl configuration. The other compound was identified as Kadsulignan L with an R- biphenyl configuration. In order to obtain kinetic data on their reversible interconversion, the stability was measured at different deuterated solvents such as deuterated methanol, deuterated chloroform and deuterated dimethylsulfoxide. The lignans were more unstable and converted more easily in deuterated methanol than in deuterated chloroform and deuterated dimethylsulfoxide.

黑老虎叶提取物纯化及其在化妆品原料中的应用研究

目的:探讨黑老虎叶提取物在化妆品原料开发中的潜在应用。方法:以黄酮含量为评价标准,构建了大孔吸附树脂对黑老虎叶提取物的纯化流程,并结合动物实验、细胞实验及化学实验,开展提取物的急性毒性及其功能特性研究。结果:AB-8大孔吸附树脂对黑老虎叶黄酮具有良好的吸附和解吸性能;上样条件为:总黄酮浓度1.0 mg/mL的料液调节pH 3,以3 BV/h流速上样5 BV;洗脱条件为:用80%乙醇以3 BV/h流速洗脱5 BV;动物实验表明,提取物无急性毒性;细胞实验显示,在2.5~50.0μg/mL浓度范围内,提取物能显著减少H_(2)O_(2)引起的人皮肤成纤维细胞死亡,有效降低细胞中ROS、MDA水平,增强SOD、CAT活性;在0.1~2.0 mg/mL浓度范围内,提取物能显著降低小鼠B16F10细胞中TYR活性;提取物抑制HAase的IC_(50)为4.748μg/mL。结论:利用AB-8大孔吸附树脂纯化的黑老虎叶提取物没有表现出急性毒性,同时展现了抗衰老、美白、抗过敏等功效,在作为化妆品原料应用上具有一定开发潜力。

基于网络药理学研究瑶药黑老虎三萜类成分对类风湿关节炎的作用机制及试验验证

目的基于网络药理学和实验研究探讨瑶药黑老虎Kadsura coccinea(Lem.)A.C.Smith治疗类风湿关节炎(rheumatoid arthritis,RA)的作用机制。方法采用硅胶、C18反相柱色谱及半制备高效液相分离技术对黑老虎的有效成分进行分离纯化;采用波谱技术对分离得到的化合物进行结构鉴定;通过Swiss Target Prediction数据库筛选黑老虎成分靶点,在GeneCards数据库和MalaCards数据库挖掘RA相关靶点,进而筛选黑老虎作用于RA的靶点;制作蛋白质-蛋白质相互作用(protein-protein interaction,PPI)网络取其交集,并通过GO分析和KEGG富集分析;采用MTT试剂盒检测化合物对肿瘤坏死因子(tumor necrosis factor,TNF)-α和白细胞介素(interleukin,IL)-6的释放情况,以及蛋白免疫印迹实验对预测结果进行验证。结果从黑老虎中提取分离出6个三萜类化合物,分别鉴定为heilaohuacid A(1),heilaohuacid G(2),seco-coccinic acid F(3),seco-coccinic acid G(4),schisandronic acid(5)和schisanlactone B(6)。MTT实验结果是化合物2,3,4,6能抑制RA-FLS细胞增殖,其IC50值为7.52~8.85μmol。6个化合物在浓度为10μmol和20μmol时均能抑制炎症因子TNF-α和IL-6的释放(P<0.05,P<0.01)。网络药理学预测得到109个共同作用靶标,通过与691个RA疾病相关靶标进行交集映射,获得其干预RA靶标38个,主要为丝氨酸/苏氨酸蛋白激酶、TNF、丝裂原活化蛋白激酶1、前列腺素内过氧化物合酶2、丝裂原活化蛋白激酶14等。GO分析和KEGG富集分析得关键通路为白细胞介素(interleukin,IL)-17信号通路和TNF信号通路等。化合物3在浓度为20μmol能上调核因子κB抑制因子α(recombinant inhibitory subunit of NF kappa B-α,IκB-α)的表达水平(P<0.05)。结论本研究从网络药理学角度预测瑶药黑老虎三萜成分治疗RA的作用机制,初步证实其可上调IκB-α蛋白的表达,为瑶药黑老虎治疗RA提供了科学依据。

黑老虎种质资源与分子生物学研究进展

黑老虎是集药用、食用和观赏于一体的功能性林下经济植物。深入研究和评价黑老虎的不同种质资源,是保护其生物多样性、拓展其资源利用与加快其品种创新的关键。本文重点综述了黑老虎不同资源(含野生和栽培种)特性研究现状,叶绿体全基因组分析、分子鉴定与遗传多样性分析、功能基因研究和实验体系建立方面的研究进展,探讨了黑老虎遗传资源研究中存在的问题和分子生物学研究面临的挑战,并指出了今后的重点研究方向。

不同品系黑老虎田间节肢动物的群落结构与多样性

为探究不同品系黑老虎田间节肢动物群落的结构和多样性,本研究以黑老虎虎绿1号、粉红1号和紫黑1号3个品系为研究对象,采用网捕、糖醋液诱集和黄、蓝板诱集等方法,于2021年对林下药用植物应用技术湖南省工程研究中心黑老虎科研示范基地田间节肢动物进行了系统调查,调查期间共收集到节肢动物6441头,隶属于4纲15目54科67种。研究结果表明,尽管不同品系黑老虎田间节肢动物各类群多样性表现不一,但差异不显著;不同品系黑老虎田间节肢动物群落的多样性指数与节肢动物总类群的变化基本一致;黑老虎田间节肢动物群落以害虫为主导,其变化趋势决定了黑老虎田间整个节肢动物群落的变化趋势,而天敌昆虫有紧随其后伴随害虫的变化而变化的现象。黑老虎田间节肢动物群落结构总体较为稳定,不同品系黑老虎田间节肢动物群落在同一地域、同一时期、相同生长环境下差异不明显。

药食两用植物黑老虎组培快繁体系研究

为改变药食同源植物黑老虎野生资源日趋减少、难以满足市场需求的现状,对组培快繁体系中的培养基配制及组培瓶苗培育条件进行了研究,以期建立黑老虎组培快繁体系,为规模化种苗繁殖奠定基础。结果表明,以MS+2.0 mg/L 6-BA+0.2 mg/L NAA为初代培养基,以B5+2.0 mg/L 6-BA+0.2 mg/L NAA为增殖培养基,以1/2 MS+0.1 mg/L 6-BA+1.5 mg/L NAA+0.3%活性炭为生根培养基,先暗培养,再光照培养,生根率可达100%,且小苗主根粗,长势强。组培苗最佳炼苗时间为15 d,最佳移栽基质为塘泥∶蔗渣∶鸡粪∶过磷酸钙=3.9∶4∶2∶0.1,最佳遮阳模式为半遮阳,最佳培育期为60 d。

广东黑老虎炭疽病病原鉴定、生物学特性及其室内药剂毒力检测

[目的]明确黑老虎炭疽病原菌种类,筛选出适宜的防治药剂,为黑老虎炭疽病的识别和大田防治提供理论依据。[方法]采用组织块分离法对广东省云浮市黑老虎炭疽病原菌进行分离,基于科赫法则确定其致病菌,采用形态学和分子生物学相结合的方法对菌株进行种类鉴定,并采用菌丝生长速率法对其进行生物学测定及不同药剂的室内毒力检测。[结果]通过对病叶分离纯化共获得6种菌株,选取具有代表性的典型菌株A1进行形态鉴定以及ITS序列分析,确定黑老虎炭疽病原菌属炭疽菌属(Colletotrichum sp.)。通过对菌株A1进行生物学测定,明确其最适生长培养基为PDA培养基,最适生长温度28℃,菌株致死条件为53℃水浴处理10 min,菌丝最适宜生长pH为6.0,最适光照条件为连续黑暗(暗24 h),最佳碳源、氮源分别为麦芽糖和酵母粉。同时,在福美双、溴菌腈、多菌灵和嘧菌酯4种供试药剂中,多菌灵的抑菌效果最佳,半数效应浓度(EC50)值为0.14μg/mL;其次是福美双和溴菌腈,EC_(50)值分别为55.26和55.32μg/mL,抑菌效果较好;嘧菌酯的EC_(50)值为791.95μg/mL,EC_(50)值在100~1000μg/mL,抑菌效果不明显。[结论]广东省云浮市黑老虎炭疽病原菌为炭疽菌属。多菌灵、福美双及溴菌腈对黑老虎炭疽病原菌菌丝生长具有较好的抑制作用,可作为黑老虎炭疽病的候选田间防治药剂。

基于CiteSpace的黑老虎研究现状与热点可视化分析

为了解黑老虎的研究现状与热点,基于中国知网(CNKI)学术期刊数据库和Web of Science(WOS)核心合集数据库,筛选出177篇中文文献和96篇英文文献,运用CiteSpace对黑老虎研究的中、英文文献的发文量、作者、机构、国家(地区)、关键词等进行可视化分析。结果表明:黑老虎研究的发文量呈上升趋势,且中文文献多于英文文献;黑老虎研究的中文文献作者形成了以梁忠厚、李有清、周玮等为核心的研究团体,英文文献作者形成了以Sun Han Dong、Pu Jian Xin、Wang Wei和Li Bin等为核心的研究团体;湖南环境生物职业技术学院园林学院和中国科学院(Chinese Academy of Sciences)分别是发表中、英文文献数量最多的机构;中国是黑老虎研究发文量最多的国家;黑老虎中文文献倾向于研究黑老虎的整体开发利用,更注重实践应用;英文文献侧重于研究化学成分及其药理作用与机制,更倾向于理论研究。

基于SSR标记的黑老虎种质资源的遗传多样性和遗传结构

【目的】揭示黑老虎5个群体70份种质资源的遗传多样性水平和遗传结构状况,为种质资源评价、保护及利用提供理论依据。【方法】用筛选出的17对引物进行所有样本SSR-PCR扩增,扩增产物经毛细管电泳后采用GenAlex v6.502、Ntsys V2.2及Structure 2.3.4等软件进行遗传多样性和遗传结构分析。【结果】17对引物的70份样本中共检测到147个等位基因,平均每对引物扩增出8.647个等位基因。17个位点的Shannon信息指数(I)平均值为1.421,期望杂合度(He)平均值为0.648,多态信息指数(PIC)平均值为0.612。5个群体中马关群体(MGH)的遗传多样性最高,I和He分别为1.070和0.544。群体遗传分化系数(Fst)平均值为0.323,有32.3%的变异来源于群体间。分子方差分析(AMOVA)显示群体间方差分量比为28.8%,群体间的遗传变异占总变异量的28.8%。17个位点群体内近交系数(Fis)的平均值为0.161(Fis>0),黑老虎存在较强的近亲交配,5个群体中只有广西群体(GXH)Fis<0。基因流(Nm)为0.876,属中等水平。UPGMA聚类和STRUCTURE分析将70份黑老虎种质资源划分成了3个类群:类群Ⅰ由来自MGH的样本资源组成,类群Ⅱ主要由屏边群体(PBH)的样本资源组成,类群Ⅲ主要由GXH、贵州群体(GZH)及湖南群体(HNH)的样本资源组成。【结论】黑老虎种质资源具有丰富的遗传多样性,群体间存在极高的遗传分化,遗传变异主要来源于群体内的个体。大部分种质的遗传背景比较单一,同一群体的不同种质遗传组分基本相同。在黑老虎育种及资源保护和开发工作中,应在尽量涵盖全部群体类别的基础上,特别重视遗传多样性丰富的群体以及各群体内不同类型个体的选择、保存和利用。