Immunoregulatory polysaccharides from Apocynum venetum L.flowers stimulate phagocytosis and cytokine expression via activating the NF-κB/MAPK signaling pathways in RAW264.7 cells

Two immunomodulatory polysaccharides(Vp2a-Ⅱ and Vp3) were isolated and identified from Apocynum venetum L. flowers, and their innate immune-stimulating functions and working mechanisms were evaluated in RAW264.7 cells. Both the level of released nitric oxide(NO) and expression of inducible nitric oxide synthase(iNOS) m RNA were significantly enhanced in the RAW264.7 macrophages cells treated by Vp2a-Ⅱ and Vp3. Vp2a-Ⅱ(100–800 μg/m L) and Vp3(400 μg/mL) could significantly increase the phagocytic activity of RAW264.7 cells and the secretion and m RNA expression of TNF-α and IL-6 in a concentrationdependent manner through affecting mitogen-activated protein kinase(MAPK) activity and nuclear factor κB(NF-κB) nuclear translocation. Vp2a-Ⅱ might activate the MAPK signaling pathways and induce the nuclear translocation of NF-κB p65, whilst Vp3 likely activated the NF-κB and MAPK signaling pathways without influencing the p38 MAPK route.

Feasibility and Prospect about the Cultivation of Apocynum venetum L. in Coastal Salt-affected Soils

[Objective]The aim of this paper was to analyze the feasibility and prospect of Apocynum venetum L.cultivation in coastal salt-affected soils.[Method]The habitat and distribution of Apocynum venetum L.in coastal area of north China were studied.A method to integrate the charicteristics of Apocynum venetum L.（Luobuma） with the utilization of salt-affected soils in this region was proposed.[Result]The introduction,domestication and growth of plants on coastal salt-affected soils can realize the protection of endangered wild species,and achieve the purpose of improving soil.The characteristics of Apocynum venetum L.can produce optimum economic and ecological benefits.[Conclusion]The implementation will provide references for the protection and exploitation of wild plant,and it is of positive significance for the integration of soil and plant resources.

Apocynum venetum Feed Affects the Serum Metabolism of Tan-sheep through the Amino Acid and Glycolipid Metabolic Pathways

[Objective]The paper was to explore the effects of Apocynum venetum diet on nutritional metabolism of Tan sheep.[Method]Forty Ningxia Tan sheep were randomly divided into 4 groups(A,B,C and D),with 10 sheep in each group.The sheep were fed with different contents of A.venetum granule feedstuff(0%,5%,10%and 15%).Blood samples were collected from jugular vein on the 20th,40th and 60th day of the experimental period.Serum samples were prepared and analyzed for differential metabolites by UPLC-Q-TOF/MS technique and annotated to the KEGG pathway.[Result](1)The differential metabolites increased with the extension of feeding time.The up-regulated metabolites in Tan sheep serum were more than the down-regulated ones,and reached the maximum on the 40th day.The up-regulated differential metabolites included 2 amino acids(valine and alanine),7 organic acids(β-hydroxybutyric acid,fumarate,ethylmalonic acid,hydroxyphenylacetic acid,gentiolic acid,protocatechuic acid,and oxycholic acid),pyrocatechol and taurochenocholate.The down-regulated differential metabolite was only p-chlorophenol.(2)With the increase of A.venetum content in pelleted diet,the differential metabolites in the serum of Tan sheep also increased,and the serum metabolic level gradually stabilized after the 10%concentration level.Similarly,the up-regulated metabolites were far more than the down-regulated metabolites.The up-regulated differential metabolites included 3 organic acids(β-hydroxybutyric acid,oxycholic acid and gentiolic acid),2 amino acids(alanine and valine)and catechol,and the down-regulated differential metabolite was only taurochenocholate.(3)The metabolic pathways involving in differential metabolites were mainly tricarboxylic acid cycle,tyrosine metabolism,taurine and bile acid pathway.[Conclusion]The results will provide a scientific basis for the green breeding of Tan sheep.

Influence on the Quality of Apocynum Venetum with Different Degumming Methods

The biochemical degumming and chemical degumming were employed to process apocynum venetum, and indices values for the residual gum content, fineness, strength of single fiber, average length, moisture regain and whiteness of degummed apocynum venetum fibers are evaluated with fuzzy mathematical method. The results obtained from this investigation show that the general index value of degummed apocynum venetum fibers with biochemical degumming （K1= 0.626 09） is prior to that with chemical degumming（K2=0.421 64）.

Protective effect of Apocynum venetum (罗布麻) against lipid peroxidation damage of erythrocyte membrane

Objective: To investigate the protective effect of Apocynum venetum on lipid peroxidation damage of erythrocyte membrane. Methods: Model of lipid peroxidation of erythrocyte membrane was manufactured by three kinds of radicals generation systems. To set up a normal control group (NC),a model control group (MC),and four Apocynum venetum groups(A.V). Observation was made on the content of malondialdehyde (MDA). Results: Compared with MC group,all of the four Apocynum venetum groups dose-dependently inhibited the increases of MDA content in the membrane induced by xanthine - xanthine oxidase system,H 2O 2 or UV light. Conclusions: Apocynum venetum may protect erythrocyte membrane from the lipid peroxidative damage induced by radicals.

盐处理对罗布麻(Apocynum venetum)幼苗渗透调节的影响

罗布麻(Apocynum venetum)是一种极具耐盐性的生态、经济、药用植物,在耐盐生理,特别是渗透调节机制方面开展研究是对其进行深度开发利用的基础。为研究盐处理对罗布麻幼苗渗透调节的影响,本试验用不同盐浓度(0,50 mmol/L,100 mmol/L,200 mmol/L NaCl)对罗布麻幼苗进行胁迫处理后,测定了无机渗透调节物质Na+、K+离子及脯氨酸、可溶性糖、甜菜碱等有机渗透调节物质的含量。结果表明:在盐处理下罗布麻幼苗中虽然Na+含量明显增加,但是K+的含量却始终能够保持相对稳定;脯氨酸、可溶性糖、甜菜碱有机渗透调节物质的含量随盐浓度的升高而显著增加。由此推测,罗布麻幼苗可以通过维持K+含量的相对稳定,并积累有机渗透调节物质来适应盐渍环境。本研究为盐渍环境下罗布麻的渗透调节机制提供一定的理论基础。

荒漠生境罗布麻(Apocynum venetum)传粉生物学比较

罗布麻(Apocynum venetum)是中国西北干旱地区的重要优势植物种,为了解其在荒漠生境的传粉生物学特性,对开花物候、花粉活力、柱头可授性、访花者种类、座果率及繁育系统进行观测。结果表明:(1)在荒漠-绿洲过渡带和荒漠边缘种群中,罗布麻的开花高峰期和花粉活力有差异,荒漠-绿洲过渡带种群的持续时间更长。(2)在过渡带种群中,姬蜂(Ichneumonsp.)是最有效的访花昆虫;在荒漠边缘种群中,意大利蜂(Apis mellifera)是主要的访花者。(3)过渡带上罗布麻种群的坐果率高于荒漠边缘。(4)在2个生境下,罗布麻杂交指数均为2,表明其繁育系统以兼性自交为主,异交在繁育系统中只起辅助作用。

An optimization of ultra-sonication-assisted extraction from flowers of Apocynum venetum in targeting to amount of free amino acids determined by UPLC-MS/MS

Amino acids are naturally occurring compounds in many edible or medicinal plants,which possess a variety of nutraceutical effects in human.Here,a method of ultrasound-assisted extraction was optimized using Box-Behnken design of response surface methodology in maximizing the yield of free amino acids deriving from flowers of Apocynum venetum L.Under the optimal condition of ultrasound-assisted extraction,i.e.187.5 W of ultrasonic power,31.93 min of extraction time,and 0.47 mg/ml of solid-liquid ratio,the experimental yield of extractive was 287.17±0.205 mg/g,which was in close agreement with the value,as predicted by the established model.In addition,a hydrophilic interaction ultra-high performance liquid chromatography coupled with QqQ-MS/MS method was developed and validated for simultaneous quantification of 20 types of amino acids without derivatization contained in A.venetum flowers.The analytical method was validated by matrix effect,linearity,limit of detection,limit of quantification,precision,repeatability,stability,and recovery.The analysis results showed that A.venetum flower was rich in free amino acids of~3%of total dried weight,and which contained leucine(13.7 g/mg),isoleucine(7.9 g/mg),and lysine(2.2 g/mg)as the most abundant amino acids.Thus,A.venetum flower could provide beneficial nutrient values for human health,e.g.adult weight maintenance or glucose homeostasis.

Ultrasound-assisted extraction of bound phenolic compounds from the residue of Apocynum venetum tea and their antioxidant activities

Bound phenolic compounds from the residue of Apocynum venetum(A.venetum)tea were released by alkaline digestion with sonication,followed by extraction using ethyl acetate(EtOAc).The fragment ions of compounds in free and bound forms were identified by liquid chromatography electrospray ionization tandem mass spectrometry(LC-ESI-MS/MS)in both positive and negative modes.Further investigation of chemical constituents in the EtOAc extract led to the isolation of eleven individual compounds,and their chemical structures were identified as(−)-epicatechin(1),(−)-loliolide(2),(−)-syringaresinol-4-O-β-D-glucopyranoside(3),alloside of benzyl alcohol(4),3-[(6-O-hexopyranosylhexopyranosyl)oxy]-2-(palmitoyloxy)propyl(9Z,12Z,15Z)-9,12,15-octadecatrienoate(5),apocynoside I(6),quercetin-3-O-β-D-glucopyranoside(7),kaempferol-3-O-β-D-glucopyranoside(8),[1-acetyloxy-3-[3,4,5-trihydroxy-6-[[3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]oxan-2-yl]oxypropan-2-yl]hexadecanoate(9),1-stearoyl-2-palmitoyl-sn-glycerol(10),and 3-hexene-l-O-β-D-glucoside(11)using a series of spectroscopic methods.Compounds 2–5 and 9–11 were isolated for the first time from A.venetum,while antioxidant experiments showed that compounds 1,7,and 8 displayed strong DPPH and ABTS+radical scavenging activities.This study represents the first investigation of bound phenolic compounds with potential health benefits in the residue of A.venetum tea.

基于代谢组学的两种罗布麻叶片类黄酮和酚酸成分分析

目的:类黄酮和酚酸物质是罗布麻中主要的活性物质,鉴定不同罗布麻品种中类黄酮和酚酸代谢物成分并比较代谢物含量差异,为有效利用罗布麻属药用资源提供理论依据。方法:以罗布麻和大麻状罗布麻叶片为材料,利用液相色谱质谱联用技术(LC-MS)检测次级代谢物成分并进行分析。结果:2种罗布麻中共检测出275种类黄酮和194种酚酸物质;PCA分析表明2种罗布麻的代谢物种类相似,但含量存在较大差异,结合OPLS-DA筛选出124种显著差异代谢物,其中有80种类黄酮和44种酚酸;富集分析发现,共有10条代谢通路被显著富集,主要体现在苯丙烷生物合成、类黄酮生物合成以及黄酮和黄酮醇生物合成等途径。结论:2种罗布麻药用成分种类相似,可为大麻状罗布麻作为药用植物开发提供理论依据。

罗布麻秆中纤维素提取及其气凝胶的制备

为实现罗布麻秆的高附加值利用,以罗布麻秆为原料,选用硝酸/乙醇法提取纤维素,通过单因素试验和正交试验优化的提取工艺为:温度85℃、时间1.5 h和固液比1∶25,提取的纤维素纯度可达98.61%。采用TEMPO氧化与超声处理相结合,制备纳米纤维素;再以其为骨架,加入聚乙烯醇和钠基蒙脱土制备纤维素复合气凝胶(NMPC)。结果表明:与纤维素气凝胶(PC)相比,NMPC对亚甲基蓝(MB)的吸附量最高达265.84 mg/g。当应变为60%时,NMPC的应力是PC的2倍。钠基蒙脱土的加入改善了气凝胶的结构,不仅提高了气凝胶的吸附性能,还提升了其压缩力学性能。

罗布麻群落特征及其与气候因子的关系

罗布麻(Apocynum venetum)是我国重要的野生种质资源,研究罗布麻群落特征及其与环境因子的关系,对罗布麻种质资源保护和合理开发利用具有重要意义。该研究以分布于甘肃省金塔县、民勤县、内蒙古杭锦旗、扎鲁特旗和吉林省白城市的野生罗布麻为研究对象,以其群落组成和生境特征为研究基础,分析生物多样性及其与气候因子的关系,解析影响罗布麻群落变化的环境因子。结果表明:不同地区罗布麻样地生物多样性存在差异,5个样地物种多样性由高到低依次为扎鲁特旗、民勤县、白城市、金塔县、杭锦旗,扎鲁特旗生物多样性最高(H=2.591,D=2.511,E=0.796,R_(1)=44.833,R_(2)=6.423),杭锦旗生物多样性最低(H=1.173,D=0.915,E=0.564,R_(1)=3.641,R_(2)=1.968)。罗布麻群落物种多样性主要与降水、温度和蒸发量密切相关,尤其丰富度指数与5—7月的降水量显著正相关,可见生长季的降水量对群落物种多样性具有重要意义。

野生罗布麻不同生境土壤理化性质及与叶片生态化学计量特征的关系

罗布麻是一种具有高抗逆性的植物,研究其土壤的理化性质和物种水平上叶片生态化学计量特征,可揭示罗布麻对环境变化的适应性。本研究在甘肃省金塔县、甘肃省民勤县、内蒙古自治区杭锦旗、内蒙古自治区扎鲁特旗和吉林省白城市野生罗布麻样地进行叶样品采集,同时采用土壤剖面法采集罗布麻根际土并进行相关分析。结果表明:5个罗布麻样地土壤肥力较低,盐碱性高,保肥性差。5个样地中,电导率最高的样地为杭锦旗样地,最低的为扎鲁特旗样地;土壤pH值最高的是白城样地,最低的是扎鲁特旗样地。Pearson相关分析表明,罗布麻叶片氮、磷含量与pH值和电导率具有显著相关性(P<0.05);叶中氮含量与土壤有机质和速效钾含量呈极显著负相关(P<0.01),相关系数为-0.606和-0.484;叶片碳含量与土壤有机质、速效钾和阳离子交换量呈显著正相关(P<0.05),相关系数分别为0.389、0.373和0.422,其与速效磷呈显著负相关(P<0.05),相关系数为-0.400。叶片磷含量与土壤有机质、速效氮、速效磷、速效钾含量以及阳离子交换量都没有显著相关性,表明叶片磷含量受土壤养分和阳离子交换量的影响较小。

盐胁迫下基于TMT技术的罗布麻蛋白质组学分析

【目的】罗布麻(Apocynum venetum L.)是中国宝贵的野生植物种质资源,具有重要的药用价值、生态价值及经济价值。虽然罗布麻是耐盐性较高的植物,但随着生境遭到破坏及土壤盐渍化程度加重,罗布麻的野生种群数量逐渐缩减。因此,探究罗布麻的耐盐性机制,对罗布麻野生种质资源的保护和利用具有重要意义。【方法】研究采用TMT技术对盐胁迫下罗布麻进行定量蛋白质组学研究,通过韦恩分析和蛋白网络互作分析方法筛选出共有差异蛋白和核心蛋白。【结果】(1)研究发现11个共有差异蛋白,主要富集在蛋白质-FAD连接、转运蛋白活性的负调控、离子跨膜转运蛋白活性的负调节、阴离子跨膜转运的负调控、阴离子通道活性的负调节和噻唑代谢过程等通路;(2)不同胁迫时间的差异蛋白网络互作所得的蛋白主要以核糖体蛋白为主,其中60S核糖体蛋白L3-2(TRINITY_DN13265_c0_g1_i1_9)在胁迫12 h和24 h的差异蛋白中均为核心蛋白,表明核糖体蛋白在罗布麻响应盐胁迫过程中发挥着重要作用。【结论】研究筛选出罗布麻响应盐胁迫的关键蛋白,拓展了罗布麻盐胁迫应答相关的分子资源,并为罗布麻天然抗逆种质的发掘与利用提供理论依据。

罗布麻研究开发利用现状及未来发展建议

本文通过查阅古今书籍文献,对罗布麻的研究开发现状及存在问题进行总结,为其未来的发展提出建议,为其进一步的开发和利用提供参考。罗布麻叶为我国传统常用的中药,药用历史悠久,含有丰富药理活性成分并具有广泛的药理作用,但也存在品名与品种混乱、质量标准缺陷和开发应用有限等问题。该药除了在降压的应用较为广泛外,其他方面的研究开发应用均不够深入广泛。建议厘清混乱名称,修订药典标准,积极推动药食两用申报,重视其药品及保健品的开发,扩大种植面积,充分发挥其在造福人类事业上的重要作用。

荷叶和罗布麻总黄酮的富集纯化工艺研究

以静态吸附率和解吸率为指标,筛选出适合荷叶、罗布麻总黄酮纯化的最佳大孔树脂,再用其进行动态吸附考察;采用单因素试验和正交试验优化荷叶、罗布麻总黄酮的纯化工艺条件。结果表明:荷叶、罗布麻总黄酮纯化的最佳树脂均是AB-8型大孔树脂,荷叶总黄酮最佳纯化工艺条件为上样流速3.0 BV/h、上样质量浓度1.606 mg/mL、乙醇体积分数80%、上样量1.4 BV,在此条件下荷叶总黄酮含量为65.75%;罗布麻总黄酮最佳纯化工艺条件为上样流速1.5 BV/h、上样质量浓度2.394 mg/mL、乙醇体积分数80%、上样量1.5 BV,在此条件下罗布麻总黄酮含量为46.75%。

基于HPLC的罗布麻与白麻不同部位活性成分比较分析

目的:比较罗布麻和混淆品白麻不同部位(叶、花、茎、根)指标成分的含量,对药材进行比较分析。方法:采用高效液相色谱法(HPLC)测定罗布麻和白麻不同部位中指标成分金丝桃苷、异槲皮苷、三叶豆苷、山柰酚-3-O-芸香糖苷、紫云英苷、芦丁、白麻苷、新绿原酸、绿原酸等14种黄酮和3种酚酸类成分含量。基于各组分进行聚类、主成分分析及正交偏最小二乘法-判别分析,并比较各组样品。结果:罗布麻和白麻不同部位样品主要活性成分为绿原酸、新绿原酸、隐绿原酸、金丝桃苷、异槲皮苷、三叶豆苷等。聚类和主成分分析显示,根中主要成分类似,叶、花中黄酮与酚酸含量均较高;罗布麻和白麻的差异成分主要表现在绿原酸、山柰酚-3-O-芸香糖苷、金丝桃苷、白麻苷、山柰酚、表儿茶素、隐绿原酸、紫云英苷、三叶豆苷、异槲皮苷。结论:罗布麻与白麻主要成分差异显著,研究结果可为评价罗布麻属药材成分差异和罗布麻传统用药提供参考。

罗布麻和大麻状罗布麻UV-B光受体UVR8基因的鉴定及表达分析

在植物响应紫外线B(ultraviolet-B,UV-B)的过程中,UV-B光受体UVR8(UV Resistance Locus 8)对植物的光形态建成和生长代谢等过程具有重要调控作用。为探究罗布麻属植物UV-B光受体信息,该研究通过罗布麻(Apocynum venetum)和大麻状罗布麻(A.cannabinum)全基因组数据进行UV-B光受体UVR8的筛选与生物信息学分析,同时利用转录组数据分析UV-B胁迫处理下的UVR8基因表达模式。结果表明:(1)罗布麻有6个UVR8基因,大麻状罗布麻有5个UVR8基因,前者分布在1、7、9和11号染色体上,后者分布在1、8和9号染色体上。(2)UVR8蛋白为亲水性稳定蛋白,定位在细胞核,不存在跨膜结构和信号肽,二级结构主要由延伸链、无规则卷曲、α-螺旋和β-转角构成。AvUVR8b和AcUVR8a蛋白三级结构与拟南芥UVR8(AtUVR8)最为类似,并且与小粒咖啡(CaUVR8)和伊德斯种咖啡(CeUVR8)的亲缘关系最近。同时发现罗布麻AvUVR8b和大麻状罗布麻AcUVR8a基因和蛋白结构与AtUVR8基因及蛋白高度相似。(3)当以一定剂量UV-B(17.52 kJ·m~(-2)·d~(-1))处理两种罗布麻植株时,AvUVR8b和AcUVR8a的表达量上调。据此推测在响应UV-B时,AvUVR8b基因在罗布麻中起主要作用,AcUVR8a基因在大麻状罗布麻中起主要作用。(4)顺式作用元件分析结果表明,UVR8的表达受光照、温度、水分、氧气和激素等因素的调控。该研究将为进一步研究罗布麻属UVR8的基因功能奠定基础,同时为解析罗布麻属植物适应UV-B的分子机制提供线索。

基于网络药理学探究罗布麻治疗非酒精性脂肪肝的作用机制

目的:基于网络药理学方法探究罗布麻治疗非酒精性脂肪肝的作用机制。方法:检索中药系统药理学数据库及文献获取罗布麻活性成分及靶点,通过GeneCards、OMIM、TTD等数据库获取非酒精性脂肪肝病相关靶点。借用String数据库对罗布麻与疾病的共同靶点构建PPI网络;利用Cytoscape软件对共同靶点进行分析以得到罗布麻治疗非酒精性脂肪肝核心作用靶点,再利用DAVID数据库对共同靶点进行富集分析。结果:筛选得到罗布麻治疗非酒精性脂肪肝核心成分20个,核心靶点包括TP53、SRC、HSP90AA1等45个,涉及癌症、脂质和动脉粥样硬化、前列腺癌、乙型肝炎、AGE-RAGE等信号通路。结论:揭示了罗布麻治疗非酒精性脂肪肝的多成分、多靶点、多通路作用特点,为深入研究罗布麻缓解非酒精性脂肪肝病的作用机制奠定基础。

罗布麻叶配方颗粒质量控制研究

目的本研究主要通过对罗布麻叶配方颗粒有效成分进行鉴别及测定,建立罗布麻叶配方颗粒质量标准。方法薄层色谱法(TLC)条件:以甲醇和水饱和正丁醇为提取溶剂,三氯甲烷-甲醇-水-甲酸(7∶2.5∶0.1∶0.2)下层溶液为展开剂,3%三氯化铝乙醇溶液为显色剂,紫外灯(365 nm)下检视;高效液相色谱法(HPLC)条件:Kromasil 100-5-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-0.2%磷酸(15∶85),检测波长256 nm,进样量10μL,柱温30℃,流速1.0 mL/min。结果对3批样品进行鉴别及测定,薄层色谱斑点清晰;液相色谱方法可靠,可用于罗布麻叶配方颗粒中金丝桃苷的含量测定。结论该方法准确可靠,可为罗布麻叶配方颗粒质量控制提供科学依据。