Tissue Culture and Rapid Multiplication Techniques of Apocynum L.

[Objective] This study aimed to investigate rapid multiplication of Apocynum by tissue culture so as to provide plantlet sources for its industrialized cultivation. [Method] The asepsis seedlings were obtained by dealing with Apocynum seeds. Its cotyledons, hypocotyls and shoot tips were cultured on the media containing different concentrations of hormones. Finally, the influence of different hormone combinations on differentiation of cotyledons and hypocotyls, rapid multiplication of shoot tips, rapid multiplication of regenerated shoots, and rooting of test-tube plantlets was com- pared. [Result] MS＋2.0 mg/L BA＋0.03 mg/L NAA and MS＋0.07 mg/L NAA were the optimum medium for inducing regenerated buds from cotyledons and hypocotyls re- spectively; MS＋2.0 mg/L BA＋0.02 mg/L NAA was the best medium for rapid multi- plication of shoot tips; MS＋1.9 mg/L BA＋I.7 mg/L NAA was the best medium for rapid multiplication of regenerated buds： and 1/2MS＋0.6 mg/L NAA was the best medium for inducing roots. [Conclusion] The optimum hormone combination was de- termined for Apocynum rapid multiplication by tissue culture, which provides technical support on Apocynum industrialized cultivation.

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.

Mechanical and Thermal Properties of Apocynum and Ramie Fiber Mat Reinforced Polylactic Acid Composites

With the increasing awareness of environmental protection and rational utilization of resources,natural fiber reinforced composites have shown broad development prospects.Apocynum fiber,known as the“king of wild fiber”,not only has moisture absorption,air permeability,and good mechanical properties but also has many health-related advantages such as antibacterial properties.In this study,four types of needle-punched Apocynum fiber and ramie fiber mat reinforced polylactic acid(PLA)composites were fabricated.Mechanical and thermal properties of the composites were tested and analyzed.The results showed that compared with those of the ramie fiber finish needle-punched mat reinforced composites,the tensile strength and the tensile modulus of Apocynum fiber finish needle-punched mat reinforced composites had increased by 15.3%and 60.1%,respectively.In comparison,the bending strength and the bending modulus were decreased by 21.8%and 7.6%,respectively.Moreover,compared with the Apocynum fiber finish needled-punched mat reinforced composites and the ramie fiber finish needle-punched mat reinforced composites,the Apocynum 50/ramie 50 finish needle-punched mat reinforced composites had the best tensile and bending properties.The after-fracture morphology was detected by a scanning electron microscope(SEM).The thermal properties of the composites were also characterized.It was found that the thermal properties of the four types of composites showed very similar behaviors.

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）.

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.

Impact of Hydrogen Peroxide/Thiourea Redox Pretreatment on the Performances of Degummed Fibers for Apocynum Degumming

The degnmming pretreatment mechanism was investigated to improve degnmming performance and the spinnability of degummed fibers. Apocynum was treated with redox system in the degumming pretreatment process and the pretreatment parameters had been optimized, besides the effects of pretreatment on property index of degnmmed apocynum fiber were investigated. The results showed that the redox system degumming process reduced alkali boiling time and had good advantages over the traditional degumming process in extraction rate, single fiber length and strength. SEM was used to test the longitudinal morphology changes of fibers before and after pretreatment. At the same time, pretreatment solution was analyzed by UV spectra and electron paramagnetic resonance spectrometer （ESR）, and its pH value was also tested. The results showed that the pretreatment solution could release acid constantly to keep the stability of the pH which caused the swelling and dissolution of pectic substances in apocynum bast. Meanwhile, pretrcatment solution could activate the free radicals from apocynam bast and reduce the activation energy of non- cellulosic composition in apocynum bast.

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.

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.

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。

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。

Study on chemical constituents of Apocynum venetum L. leaves by LC/MS and determination of the best harvest season

The dried leaves of Apocynum venetum L. （AVL） is a traditional Chinese medicine widely used as antihypertensive medicine in China. Accurate determination of its best harvest season is important for its effective and safe use. In the present study, we developed a reliable method based on high-performance liquid chromatography-diode array detector-electro spray ionization-ion trap-time of flight （HPLC-DAD-ESI-IT-TOF） mass spectrometry and HPLC-DAD for the identification and quantification of major components in AVL leaves. Quantitative analysis of 24 samples collected weekly helped monitor the changes of compounds dynamically in AVL leaves to determine the best harvest season. A total of 30 compounds were identified, including quinic acid, five phenolic acids and 24 flavonoids. For the first time, 16 compounds were selected as marker compounds and simultaneously monitored weekly instead of monthly during the growth of the plant. The results showed that in May the leaves had the highest amount of phenolic acids, flavonoids and total compounds. Therefore, May should be the best harvest season for AVL leaves, which was distinct from previous studies. The established method was validated to be simple, accurate and precise, and thus it was of great importance for determination of the best harvest season.

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.

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.

A STUDY ON FORMATION OF BAST FIBERS IN THE STEM OF APOCYNUM LANCIFOLIUM RUS

Apocynum lancifolium Rus is a kind of shrub-like herbaceous plant belonging to Genus Apocynum L. of Family Apocynaceae. It is widely distributed and grown abundantly in China and considered as a resource plant for various usage, such as spinning and weaving, paper-making and drug-manufacturing.

罗布麻杆提取木质素的气凝胶性能

选择氯化胆碱(ChCl)与乳酸(LA)合成低共熔溶剂(DES)来提取罗布麻秸秆中的木质素,并将提取的木质素(DL)与聚乙烯醇(PVA)共混,以四硼酸钠为交联剂,通过冷冻干燥技术制备了气凝胶A-PBDL。试验结果表明:DES提取木质素最佳工艺为:n(ChCl)∶n(LA)=1∶9,120℃提取9 h;红外图谱分析得知木质素中含有对羟基苯丙烷、创愈木基和紫丁香基三种结构,平均粒径为3.601μm。研究了气凝胶制备中木质素添加量、PVA浓度和四硼酸钠浓度对气凝胶孔隙率和强力的影响,发现掺杂少量木质素可显著提高气凝胶的力学性能及孔隙率,并增强其热稳定性。

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。

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。

罗布麻苷提取和检测条件优化及其年限累积效应分析

为优化罗布麻中罗布麻苷的提取和检测条件,明确种植年限对罗布麻苷累积的影响,本试验利用超声波法提取罗布麻苷,用3,5-二硝基苯甲酸-氢氧化钠显色法测定其含量,用铁离子还原/抗氧化能力法(FRAP)和1,1-二苯基-2-三硝基苯肼(DPPH)法检测其抗氧化能力。结果表明:罗布麻苷标准品显色反应液在540 nm下的吸光度与其含量呈良好的线性关系,线性范围为0~1.0 mg/mL(R^(2)=0.992),样品回收率为99.218%,相对标准偏差(RSD)为1.909%。在考察范围内,对提取试剂乙醇浓度、超声时间和料液比进行分析发现,待测样品与95%乙醇的料液比为1∶10(g/mL)、超声提取30 min时,罗布麻苷含量最高,达到639.34μg/g。种植3、7、10年的罗布麻不同部位罗布麻苷含量差异显著,依次为7年叶>3年叶>10年叶>3年茎>7年根>10年根>3年茎>7年根>10年茎。年限间比较,种植7年的罗布麻具有较高的罗布麻苷含量,为626.25μg/g,抗氧化能力表现为7年>3年>10年,且3个种植年限间均有较强的DPPH自由基清除能力。相关性分析表明,种植7年的罗布麻中罗布麻苷含量与抗氧化能力呈正相关。本研究结论可为罗布麻苷提取及相关功能性产品开发提供依据。

罗布麻黄酮提取及纯化方法研究进展

为了促进罗布麻黄酮活性成分在纺织领域的开发与应用,文中列举了罗布麻中主要的黄酮类成分,详细阐述了罗布麻总黄酮的提取及分离纯化方法。提取方法包括:溶剂提取法、超声波辅助提取法、超临界流体萃取法、酶辅助提取法、微波辅助提取法及离子液体提取法;分离纯化方法包括:柱色谱法和分配色谱法。同时分析比较了各提取和纯化方法的作用原理及优缺点,指出罗布麻黄酮提取纯化方法未来发展方向。

罗布麻叶不同极性部位及成分对药物诱导的斑马鱼心脏损伤的保护作用研究

目的 基于斑马鱼模型评价罗布麻叶提取物及主要化学成分对药物诱导心脏损伤的保护作用。方法 用野生型AB斑马鱼测试半数致死浓度(LC50);采用盐酸维拉帕米构建斑马鱼心脏损伤模型,并评价不同极性部位及主成分对斑马鱼心包水肿面积和静脉淤血面积的影响。结果 在安全剂量下,罗布麻叶Fr.n-BuOH、Fr.n-BuOH-40%以及Fr.H2O部位对斑马鱼心脏损伤具有更好的保护作用;主成分金丝桃苷的保护作用呈现明显的剂量相关性,且略强于异槲皮苷。结论 罗布麻叶提取物及其主成分对斑马鱼心脏损伤具有保护作用,值得进一步开展机制研究。