A Novel Phenylpropanoid-substituted Catechin Glycoside and a New Dihydrochalcone from Sarcandra glabra

A novel phenylpropanoid-substituted catechin glycoside glabraoside A 1 and a new dihydrochalcone 3＇-（7＂-allylphenyl）-2＇,4＇,4＂-trihydroxy-6＇-methoxydihydrochalcone 2 were isolated from the herbs of Sarcandra glabra. Their structures were elucidated on the basis of spectroscopic analyses and chiroptical methods.

Study on chromatographic fingerprint of sarcandra glabra (Thunb.) by microwave-assisted extraction coupled to HPLC/DAD

Microwave-assisted extraction(MAE)was used for extraction of effective components of sarcandra glabra(Thunb.),and then chromatographic fingerprint of sarcandra glabra(Thunb.)was studied by high performance liquid chromatography/diode array detector(HPLC/DAD).The conditions of MAE were optimized by an orthogonal experiment,and then the authentication and validation of the chromatographic fingerprint were conducted.Nine peaks were identified as common peaks in the fingerprint chromatograms,and isofraxidin was considered as a reference compound and quantified.Relative standard deviations of retention time and peak area of each component were less than 3% and 8%,respectively.Similarity and difference analysis were conducted by use of PCA and relation coefficient.Twenty batches of sarcandra glabra(Thunb.)samples from two different producing areas could be classified into two different groups in the PCA model.The results showed that MAE-HPLC/DAD method was simple,efficient and stable for the study of complex chromatographic fingerprint of sarcandra glabra(Thunb.),which could provide more reliable and precise information for quality evaluation.

缙云山草珊瑚(Sarcandra glabra)天然居群结实特征的变异性研究

对缙云山草珊瑚8个天然居群结实特征的20项表型性状的变异性进行了研究,结果表明:各表型性状在居群内与居群间都存在很大的变异性,其中种实产量、果序数和果序分枝的变异程度较高,而果实和种子的特征指数变异程度较低,同类性状果实的变异程度要大于种子;平均变异系数最大的性状是四分枝果序百分比(1.477),最小的是种子长宽比(0.041);变异程度最大的居群是居群6(0.564),最小的是居群4(0.292);各居群间除果序分枝总数差异不显著、果序数和二分枝果序百分比差异显著外,其它17项表型指标均达到极显著水平,表明缙云山草珊瑚各居群间已经发生了明显的形态分化;居群内和居群间变异分别占总变异的60.7%和39.3%,说明居群内变异是表型变异主要来源,但居群间变异也不容忽视;欧氏距离聚类将所有居群划归为两大类群,主要与居群的群落类型与干扰程度有关;表型性状的Pearson相关性聚类显示草珊瑚同类性状在表型分化过程中具有很强的相关性。

Sowing and Seedling Experiment on Different Substrate Nutrient Bags for Sarcandra glabra

A comparative experiment was carried out on sowing and seedling for seeds of Sarcandra glabra( Thunb.) Nakai treated by different methods( A-harvesting and sowing simultaneously with episperm removed; B-harvesting and sowing simultaneously with episperm not removed) and different substrate nutrient bags( A-forest land topsoil; B-yellow soil∶ coarse sand = 1∶ 1; C-farmland soil). Results indicate that the seed germination rate of S. glabra seeds harvested and sowed simultaneously with episperm removed and not removed was as high as 97. 3%and 97. 1%,respectively; the seedling growth of sowing substrate A-forest land topsoil was the highest,the average seedling height,average ground diameter,average main root length,and average lateral root length was 12. 1,0. 21,8. 5 and 7. 0 cm,respectively.

Sarcaglarols A-D, Lindenane-Monoterpene Heterodimers from Sarcandra glabra Based on Molecular Networks

Sarcaglarols A-D(1-4),two pairs of lindenane-monoterpene heterodimers fused by a 1,2-dioxane moiety,were discovered and isolated from the leaves of Sarcandra glabra guided by MS/MS molecular networking-based strategy.Their planar structures,absolute configurations of basic skeleton and flexible polyhydric side chain were established by analysis of HRESIMS,NMR spectroscopic data,ECD spectrum,and the X-ray diffraction study of isopropylidene derivatives.An intermolecular[2+2+2]cycloaddition may play a key role in the biosynthesis pathway of the 1,2-dioxane moiety fused lindenane-monoterpene heterodimer skeleton,which can be recognized as the biogenetic precursors of our previous reported lindenane-normonoterpene conjugates.In addition,compounds 1,3 and 4 exhibited moderate inhibitory effects of lipid accumulation in free fatty acid-exposed L02 cells.

Sesquiterpenoids from the leaves of Sarcandra glabra

Sarglanoids A-F,six new sesquiterpenoids belonging to eudesmane(1-5)and eremophilane(6)types,were isolated from the leaves of Sarcandra glabra,a famous traditional Chinese medicine(TCM).Their structures including absolute configurations were elucidated through extensive spectroscopic analysis and electronic circular dichroism(ECD)calculations.Compounds 1-2 were rare N-containing eudesmane-type sesquiterpenoids.Compound 3 exhibited inhibitory activity against nitric oxide(NO)production in lipopolysaccharides(LPS)-induced RAW 264.7 cells with IC_(50) values at 20.00±1.30μmol·L^(-1).These findings provide scientific evidence for sesquiterpenoids as the material foundation of S.glabra.

Ultra-High-Performance Liquid Chromatograph with Triple-Quadrupole Mass Spectrometer Quantitation of Twelve Phenolic Components in Different Parts of Sarcandra glabra

Objective:The study objective was to determine phenolic components for the quality control(QC)of cultivated Sarcandra glabra(Thunb.)Makino(S.glabra).Materials and Methods:A sensitive,ultra-high-performance liquid chromatography-tandem mass spectrometric method for the simultaneous determination of 12 phenolic components has been developed.Six caffeoylquinic acids,two caffeoylshikimic acids,and four flavanonol glucosides were selected for the comprehensive analysis of distribution in different parts(root,stem,and leaf).Results:Twelve phenolic components were linear in the concentration range of 0.005–5.0μg/mL(R^(2)>0.995).The relative standard deviation of intra-day and inter-day precision across three validation runs over the entire concentration range was<5%.The recovery determined was within 5%in terms of relative error.Our results showed that the 12 phenolic compounds were mainly distributed in leaves and stems far more than those in roots.Conclusions:This study provided an ultra-high-performance liquid chromatograph with triple-quadrupole mass spectrometer quantitative method of 12 phenolic components for the QC of cultivated S.glabra.It was found that the phenolic components were significantly accumulated in the aerial parts(stems and leaves)of cultivated S.glabra.

Clinical observation on prevention and treatment of radiation-induced parotid-gland injury in patients with nasopharyngeal carcinoma by Compound Sarcandra Glabra

Objective:To investigate the protective effect of the Compound Sarcandra Glabra on radiation injury of parotid glands in patients with nasopharyngeal carcinoma.Methods:Sixty patients with primary nasopharyngeal carcinoma in the First Affiliated Hospital of Guangxi Medical University from June 2018 to January 2019 were collected,which were divided into treatment group and control group using a random number table.All patients were treated by a combination of 9-field intense-modulated radiotherapy and chemotherapy.The treatment group was given the Compound Sarcandra Glabra orally every day from 3 days before radiotherapy to the end.The degree of xerostomia(dry mouth)of patients was compared between the two groups at the end of radiotherapy and 3 months after radiotherapy by the chi-square test.The maximal adsorption and secretion rates of the parotid glands were compared between groups before radiotherapy,at the end of radiotherapy and 3 months after radiotherapy by the independent sample t-test.Results:The symptoms of dry mouth were alleviated in the treatment group,and the degree of dry mouth significantly differed between groups at the end of radiotherapy and 3 months after radiotherapy(χ2=6.696 and 10.977,P<0.05).There were no significant differences in the maximal adsorption and secretion rates of the parotid glands between groups before radiotherapy(t=0.974 and4.646,P>0.05);however,both variables were significantly higher in the treatment group at the end of radiotherapy and 3 months after radiotherapy(t=2.920 and 5.037;t=2.820 and 2.213,P<0.05).Patients presented no liver and kidney injury after radiotherapy.Conclusions:Oral administration of the Compound Sarcandra Glabra can effectively prevent radiation injury of the parotid glands in patients with nasopharyngeal carcinoma,thereby alleviating the degree of dry mouth after radiotherapy.

Three new compounds from Sarcandra glabra

一新 sesquiterpene 内酯， 1,8,9-trihydroxyeudesman-3 (4 ) ， 7 (11 )-dien-8,12-olide (1 ) 和二新代替 phenylpropanoid 的 catechin glycosides， glabraoside C (2 ) 和 glabraoside D (3 ) 从 Sarcandra glabra 的整个植物被孤立。他们的结构被分析建立光谱、化学证据。

草珊瑚内生真菌SgG4抗植物病原真菌的活性物质研究

对草珊瑚内生真菌卷曲木霉Trichoderma spirale SgG4抗植物病原真菌的活性物质进行研究。采用菌丝生长速率法对SgG4发酵产物进行抑菌活性测定,采用硅胶柱层析法并结合活性追踪对发酵产物的活性萃取物进行活性物质分离纯化,运用核磁共振谱和质谱技术对活性化合物进行结构鉴定。结果表明,抗菌活性物质主要存在于发酵产物的乙酸乙酯萃取物中,该萃取物对13种植物病原真菌均有较好的抑制活性,EC 50值为0.0002~2.4757 g/L,其中对甘蔗凤梨病菌的抑制活性最好。从发酵产物的乙酸乙酯萃取物中分离出1个抑菌活性化合物,鉴定为木霉酸(trichoderma acid)。木霉酸在0.1 g/L时,对13种植物病原真菌均有抑制作用,其中对茶轮斑病菌、烟草黑胫病菌、甘蔗凤梨病菌和白绢病菌的抑菌率为100%,对其余9种植物病原真菌的抑菌率为21.43%~85.71%。可见,木霉酸的抑菌活性较好且抗菌谱广,在微生物源杀菌剂方面具有较好的开发应用前景。

杉木土壤酚酸物质对草珊瑚生长和品质的影响

【目的】探究杉木人工林土壤中含量较高的4种酚酸物质对草珊瑚生长和品质的化感效应,为杉木林下套种草珊瑚提供理论依据,同时为利用其化感效应指导生产提供参考。【方法】以3个月生草珊瑚为试验材料,分别施加20,40,60,80 mg/L的对羟基苯甲酸、阿魏酸、香草酸和没食子酸处理液,以清水处理为对照,探究其对草珊瑚幼苗生长(苗高增量、地径增量、地上生物量、地下生物量)、光合生理(光合色素含量、净光合速率、蒸腾速率、气孔导度、胞间CO_(2)浓度)和药用成分(异嗪皮啶、绿原酸、迷迭香酸、总黄酮)含量的影响,利用平均敏感指数、相关性分析和主成分分析进行综合评价。【结果】①与对照相比,20~60 mg/L对羟基苯甲酸、阿魏酸和没食子酸总体上对草珊瑚的苗高增量、地径增量、地上生物量、地下生物量以及异嗪皮啶、绿原酸、迷迭香酸和总黄酮单株含量表现出促进作用,而香草酸则仅在质量浓度为20~40 mg/L时表现出促进作用。②与对照相比,4种酚酸在质量浓度为20~40 mg/L时对草珊瑚叶片的净光合速率、气孔导度和蒸腾速率表现为促进作用;在20~60 mg/L时对叶绿素a、叶绿素b、总叶绿素及类胡萝卜素含量提升表现为促进作用;在质量浓度为80 mg/L时对胞间CO_(2)浓度表现为促进作用。③综合分析可知,酚酸处理下草珊瑚的光合生理指标与生长指标、药用成分之间有较强的相关性,说明在一定阈值内酚酸物质可通过增强草珊瑚植株的光合作用,进而促进草珊瑚生长及其体内药用成分的累积。【结论】4种杉木土壤酚酸物质对草珊瑚生长和品质整体呈现出低中质量浓度(20~60 mg/L)促进、高质量浓度(80 mg/L)抑制的化感效应,且20 mg/L阿魏酸、60 mg/L对羟基苯甲酸、40 mg/L的香草酸和没食子酸在促进草珊瑚生长和品质提升方面表现较优。

九节茶药材质量标准研究

目的 对九节茶药材开展全面的质量标准研究,建立完善的九节茶质量标准。方法 收集13批九节茶药材,以异嗪皮啶对照品和九节茶对照药材建立薄层色谱鉴别方法;参考《中国药典》(2020年版)四部通则,对九节茶水分、总灰分、酸不溶性灰分及浸出物进行检查;采用高效液相色谱法测定九节茶中异嗪皮啶和迷迭香酸的含量。结果 九节茶薄层鉴别特征斑点清晰,分离度良好;13批九节茶药材水分平均为9.4%,总灰分平均为5.2%,酸不溶性灰分平均为0.5%,浸出物平均为12.6%;异嗪皮啶平均含量为0.026%,迷迭香酸平均含量为0.030%。结论 本研究建立的薄层鉴别、检查及含量测定方法专属性强、准确度高、操作简便,可用于九节茶药材质量标准研究。

草珊瑚中类胡萝卜素合成的内源激素调控机制分析

【目的】为了探究草珊瑚[Sarcandra glabra(thunb)nakai]不同器官中内源激素对类胡萝卜素差异积累的可能调控机制。【方法】通过代谢组学方法分析草珊瑚中内源激素和类胡萝卜素代谢物在不同器官间的差异分布情况,结合转录组学技术挖掘草珊瑚类胡萝卜素相关差异酶基因,进一步预测参与调控差异酶基因的转录因子及其激素相关顺式响应元件,从而分析内源激素对类胡萝卜素差异积累的可能调控作用。【结果】吲哚-3-羧酸(indole-3-carboxylic acid)、反式茉莉酸[(-)-jasmonic acid]、二氢茉莉酮酸甲酯(ethyl dihydrojasmonate)、油菜素甾酮(castasterone)、油菜素内酯(brassinolide)等7种内源激素在叶中的含量更高,与角黄素(canthaxanthin)、海胆酮(echinenone)、新黄质(neoxanthin)和念珠藻黄素(nostoxanthin)等8种差异代谢物等富集部位一致;而脱落酸(abscisic acid,ABA)、赤霉素4,5-甲氧基水杨酸(gibberellin 4,5-methoxysalicylic acid)、二氢茉莉酸(dihydrojasmonic acid)和5,6-二羟基吲哚(5,6-dihydroxyindole)在根中的含量更高,与脱落酸醛(abscisic aldehyde)、4,4'-二聚戊烯(4,4'-aiapolycopenedial)、花药黄质(antheraxanthin)这3种差异代谢物富集部位一致;关键转录因子MYB106、SPL1、NAC015、ERF064、WRKY44、BHLH116可通过响应AUX、SA、GA、ABA、Me_JA等植物激素的刺激,参与调控类胡萝卜素合成途径的DXS、VDE、ABA2、AOG、ZEP、CYP97C1、DWARF27、CRTISO、LCYB、PSY这10种代谢酶的表达。实时荧光定量PCR(quantitative real-time polymerase chain reaction,RT-qPCR)结果表明,随机选取的8个差异基因表达趋势与转录组测序结果一致。【结论】筛选出草珊瑚叶和根与类胡萝卜素相关的15 种差异内源激素与 11 种差异代谢物,预测了6种转录因子参与调控10 种代谢酶。

不同脱水条件对草珊瑚种子萌发及生理指标的影响

[目的]对不同时间梯度脱水条件下的草珊瑚种子进行萌发试验及生理生化指标测定。[方法]用硅胶脱水(快速脱水)和30℃恒温培养箱脱水(慢速脱水)2种方法获取不同含水量的草珊瑚种子。对不同含水量种子进行萌发测试,并采用紫外分光光度法对种子中各项理化指标进行测定,综合判断种子的脱水耐性及其生理基础。[结果]随着脱水时间的延长,草珊瑚种子含水量呈逐渐下降趋势。在慢速脱水24 h条件下,草珊瑚种子平均含水量为5.28%,导致种子丧失萌发能力。而快速脱水8 h的草珊瑚种子含水量已低于10%,其萌发率仅为16.67%,亦较大程度地丧失萌发能力,因此慢速脱水24 h和快速脱水8 h可作为草珊瑚种子脱水的最终梯度。快速脱水的草珊瑚种子比慢速脱水具有更高的萌发率。当种子含水量接近且低于10%时,快速脱水种子中的过氧化氢酶(CAT)活性比慢速脱水高,而过氧化氢(H 2O 2)含量和超氧阴离子含量比慢速脱水低。脯氨酸含量在慢速脱水条件下并未发现增加趋势,但快速脱水后期(24 h)显著增加。慢速脱水后期(4~24 h)种子可溶性糖含量增加,而快速脱水期间可溶性糖含量基本保持不变。在快速脱水16 h、慢速脱水24 h的淀粉含量均增加,但当种子含水量接近且低于10%时,快速脱水种子的淀粉含量比慢速脱水低。草珊瑚种子内β淀粉酶活性高于α淀粉酶。[结论]该研究为草珊瑚种子萌发条件和萌发机理提供一定的参考依据。

杉木林下套种草珊瑚对土壤养分的影响

为分析套种草珊瑚对杉木人工林土壤养分的影响,本研究以杉木林和杉木-草珊瑚套种林两个林分类型为研究对象,在2019年1、4、7、9、11月采集两种林分不同土层土壤样品,检测分析两种林分不同土层土壤样品的pH值,以及有机质、全氮、碱解氮、全磷、有效磷、全钾和速效钾等养分含量的差异。结果表明,杉木林下套种草珊瑚可以提高土壤pH值,增加土壤有机质、全氮、碱解氮、全磷、有效磷、全钾和速效钾的含量;在养分流失严重的春夏季,套种草珊瑚有利于杉木林土壤中养分的积累。杉木林下套种草珊瑚对土壤养分有一定影响,能提高土壤养分含量,有利于提高林地生态和经济效益。

氯化胆碱基低共熔溶剂优化草珊瑚总黄酮提取工艺

为优化草珊瑚总黄酮提取工艺,采用单因素试验探究了5种氯化胆碱基低共熔溶剂、氢键受体与氢键供体的摩尔比、提取时间、液料比、提取温度对草珊瑚总黄酮得率的影响,通过L9(34)正交试验优化提取工艺。结果表明:最优工艺为氯化胆碱/乙二醇摩尔比1:25,时间60 min,液料比30:1(mL/g),温度80℃,平均得率为17.20%,RSD为0.64%。低共熔溶剂较传统溶剂(水、醇)对草珊瑚总黄酮的提取率高,氯化胆碱/乙二醇体系低共熔溶剂是一种绿色高效的提取草珊瑚总黄酮的溶剂。

熵权法结合Box-Behnken响应面法优化肿节风药材的提取工艺

目的优化肿节风药材的提取工艺。方法以高效液相色谱(HPLC)法测定的肿节风中迷迭香酸与异嗪皮啶含量为考察指标,对超声时间、超声温度、液料比(mL/g)、甲醇体积分数进行单因素实验。基于单因素实验结果,采用Box-Behnken响应面法设计实验,运用熵权法赋予各指标权重并计算综合评分,再以综合评分为评价指标,优化肿节风药材的提取工艺,并对优化后的提取工艺进行验证。结果肿节风药材最佳提取工艺为超声时间40 min,超声温度45℃,液料比50∶1,甲醇体积分数70%。3次验证实验结果显示,平均综合评分为0.9886,RSD为0.50%,各综合评分实际值与预测值(0.9851)之间的偏差均在±1%以内。结论本研究优化后的工艺稳定可行、重复性好,可为肿节风药材的提取提供参考。

肿节风及其复方制剂临床应用研究进展

肿节风又名草珊瑚,含有挥发油、酯类、酚类、鞣质、黄酮、氰甙、香豆素、内酯等化学成分,具有抗炎、抗菌、抗氧化、抗肿瘤和神经保护等多种生物活性,以肿节风为原料的中药近年来在临床上得到广泛应用。综述近十年来,肿节风及其复方制剂在治疗咽喉炎、风湿性关节炎、胃溃疡、急性化脓性中耳炎、放射性肺损伤、小儿肠系淋巴结、骨外科疾病、癌症等方面的临床研究进展及现代作用机制,以期为肿节风及其复方制剂的临床合理用药及再开发利用提供参考。

杉木林下草珊瑚根系丛枝真菌群落的季节动态研究

以杉木林下草珊瑚为研究对象,通过测定其根系中丛枝菌根真菌(AMF)侵染率、根际土壤菌丝密度、根内AMF群落多样性以及土壤理化性质,探讨草珊瑚根内AMF群落结构的季节动态,以及与土壤理化性质的关系。结果表明,在草珊瑚根系中共检测到440个AMF的分类单元,分属9科14属,其中球囊霉属为绝对优势属,且1月和9月相对丰度高达99%。AMF侵染率和菌丝密度在不同月份存在波动,侵染率也是在1月和9月有两个峰值,且9月根际土壤的菌丝密度最大。土壤全磷、全钾、有效磷、速效钾和含水量是影响草珊瑚根系AMF群落结构的主要因子,其中土壤全磷和含水量影响显著(P<0.05)。

草珊瑚不同提取物抑菌活性研究

目的研究草珊瑚不同提取物的抑菌活性。方法以水作为溶剂,采用回流提取方式得到草珊瑚的水提液后,依次采用石油醚(30℃~60℃)、乙酸乙酯、正丁醇等溶剂对草珊瑚水提液进行萃取得到不同草珊瑚提取液,旋转浓缩得到相应浸膏,并用水复溶成不同浓度梯度溶液,利用琼脂平板打孔法评价不同草珊瑚提取物对小麦赤霉病菌、须癣毛癣菌、炭疽病菌、红色毛癣菌、烟草赤霉病菌、白术根腐病菌、大肠杆菌以及金黄色葡萄球菌的抑菌活性。结果在供试菌种中,草珊瑚水提液对大肠杆菌、金黄色葡萄球菌、须癣毛癣菌、烟草赤霉病菌表现出不同程度的抑制作用,进一步对草珊瑚水提液进行萃取发现,不同草珊瑚提取物抑菌活性均有一定的抑制效果,且随着不同提取物的浓度增大其抑菌效果也逐渐增强。其中,正丁醇和乙酸乙酯萃取物的抑菌活性较好,正丁醇萃取物对须癣毛癣菌最小抑菌浓度为15 mg/mL,对金黄色葡萄球菌最小抑菌浓度为15 mg/mL,对大肠杆菌最小抑菌浓度为10 mg/mL;乙酸乙酯萃取物对烟草赤霉病菌最小抑菌浓度为15 mg/mL。结论:草珊瑚提取物具有一定的抑菌活性,对大肠杆菌、金黄色葡萄球菌、须癣毛癣菌、烟草赤霉病菌具有抑菌作用,采用正丁醇和乙酸乙酯对草珊瑚水提液进行萃取的得到的提取物抑菌效果更好,草珊瑚可作为一种天然抑菌植物进一步进行开发。