Isolation and Purification of Plantamajoside and Acteoside from Plant Extract of Plantago asiatica L. by High Performance Centrifugal Partition Chromatography

Two phenylethanoid glycosides（PhGs）, plantamajoside and acteoside, were isolated and purified from the aerial parts ofPlantago asiatica for the first time by high performance centrifugal partition chromatography（HPCPC） with ethyl acetate, n-butanol, ethanol and water（0.5：0.5：0.1：1, volume ratio） as solvent system. A total of 45.6 mg of plantamajoside and 293.8 mg of acteoside were purified from 1341 mg of the n-butanol extract of P. asiatica, with a purity of 〉93.3% as determined by HPLC. The HPCPC fractions were analyzed by HPLC-DAD and the structures were identified by their retention time, UV, electrospray ionization multi stage tandem mass spectrometry（ESI-MSn） in the negative ion mode, and confirmed by NMR experiments. The characteristic fragment ions of ESI-MS of the two PhGs isolated from Plantago asiatica were discussed, which are specific and useful for the identification of the structures of PhGs.

车前(lantago asiatica L.)毛状根诱导技术研究

为建立发根农杆菌Ri质粒介导的车前(lantago asiatica L.)转化体系,通过发根农杆菌A4和R1601侵染车前无菌苗的外植体诱导毛状根.实验结果表明,车前外植体最佳侵染时间为8 min;两种菌液均可以诱导车前叶片、叶柄和根产生毛状根,车前叶片和叶柄毛状根的诱导率低于40%,车前根部毛状根的诱导率高于80%;经过分子鉴定,Ri质粒T-DNA的rolB基因已成功整合到毛状根中,表明车前的根是适宜毛状根诱导的最佳外植体,可为今后车前毛状根的扩大培养奠定基础.

浙江产车前(Plantago asiatica)种群遗传分化的主坐标分析

运用 18个有效的 10mer 寡核苷酸引物对浙江产车前 (Plantagoasiatica)的 8个种群基因组DNA进行了PCR扩增 ,共检测到 186个位点 ,其中多态位点 12 5个 ,占 6 8.70 % .应用Jaccard公式计算了 8个种群在 12 5个位点上的相似性 .以此为基础 ,应用主坐标排序 ,作出了 8个种群遗传分化的三维排序图 .结果表明 ,8个种群的遗传分化与地理位置、海拔高度有联系 .

Isolation and purification of phenylethanoid glycosides from plant extract of Plantago asiatica by high performance centrifugal partition chromatography

Two phenylethanoid glycosides （PhGs） were isolated and purified from the aerial parts of Plantago asiatica for the first time by high performance centrifugal partition chromatography （HPCPC） using ethyl acetate-n-butanol-ethanol-water （0.5：0.5：0.1：1, v/v/v/v）. A total of 45.6 mg of compound 1 and 293.8 mg of compound 2 were purified from 1341 mg of the n-butanol extract of P. asiatica. The structures of the two PhGs were tentatively identified as plantamajoside and aeteoside or isoacteoside by eleetrospray ionization multi stage tandem mass spectrometry （ESI-MS^n） in the negative ion mode.

北美车前(Plantago virginica)和车前(P.asiatica)分布格局的分形分析

Box dimension and information dimension based on individual and leaf number were applied to analyze the distribution patterns of Plantato asiatica and P. viriginica in suburbs of Jinhua. The results showed that both the box dimension and the information dimention of P. virginica were obviously higher than those of P. asiatica in the high human-distributed environment. P. asiatica was mostly distributed in the environment with compacted soil. The information dimension on the basis of leaf probability in the boxes showed that the populations of P. virginica and P. asiatica in low density could exploit environmental resources effectively.

Four new indole alkaloids from Plantago asiatica

Four new indole alkaloids,plasiaticines A-D(1-4),together with two known ones,were isolated from the seeds of Plantago asiatica.The structures of the new compounds were elucidated on the basis of comprehensive analysis of spectroscopic data.All compounds were tested for their cytotoxic activity,and all compounds except 4 were tested for their acetylcholinesterase(AChE)inhibitory activities.

Anti-fatigue Effect of Water Extract from Centella asiatica(L.)Urban on Mice

[Objectives]To study the anti-fatigue effect of water extract from Centella asiatica(L.)Urban on mice.[Methods]After intragastric administration of low,medium and high concentration of water extract from Centella asiatica(L.)Urban for 14 d,the rotarod time and the content of serum lactic acid in mice were determined,respectively.[Results]Compared with the control group,the rotarod time of mice in the low and medium concentration groups was significantly prolonged(P<0.05),but there was no significant difference between the control group and the high concentration group;the content of serum lactic acid in the medium concentration group was significantly lower than that in the control group(P<0.01),but there was no significant difference between low concentration group and high concentration group and the control group.[Conclusions]A certain concentration of water extract from Centella asiatica(L.)Urban had a good anti-fatigue effect.

车前属(Plantago L.)3种车前种子的生物学比较研究

本研究对黑龙江省车前科(Plantaignaceae)车前属(Plantago L.)的3种野生车前种子的生物学特性进行比较,为黑龙江地产药材车前属植物的引种驯化和开发利用提供依据。采用显微观察、生物学对比等方法,对种子形态和种子萌发的影响因素进行比较研究。结果表明:3种车前的种子形态存在差异,平车前的种子不存在休眠现象,通过低温和赤霉素协同处理有利于大车前和车前种子的萌发。因此,大车前和车前的种子在引种驯化前需要经过低温处理。

Investigation on the Material Basis of Sijicao Granules in Treating Eczema Based on Network Pharmacology

[Objectives]To explore the pharmacodynamic material basis of Sijicao granules for the treatment of eczema through chemical composition-network pharmacology.[Methods]First of all,the chemical constituents of Polygonum capitatum and Plantago asiatica from Sijicao granules were collected,and the relevant target information of the constituents was collected by TCMSP,PubChem,DisGeNET,GeneCards and STRING databases.Furthermore,Cytoscape 3.8.2 software was used to construct the chemical compounds-target network map of Sijicao granules.Finally,STRING database was used for PPI protein network analysis,GO functional enrichment analysis and KEGG pathway enrichment analysis of core targets,and molecular docking between core constituents and protein targets was also performed.[Results]30 constituents,including quercetin,kaempferol,luteolin,ellagic acid and gallic acid,were discovered to be the key effective compounds of Sijicao granules in the treatment of eczema.And its core action protein targets were PTGS2,NOS2,AKT1,TP53,IL6,HMOX1.What s more,through GO functional enrichment analysis of biological process(BP),cell component(CC),molecular function(MF)analysis and KEGG pathway enrichment analysis,the main pathways of action of Sijicao granules for the treatment of eczema including IL-17 signaling pathway,T cell receptor signaling pathway,cancer signaling pathway,TNF signaling pathway and Relaxin signaling pathway.In addition,molecular docking results displayed that the primary active constituents quercetin,kaempferol and luteolin were well combined with the core protein targets AKT1 and IL6.[Conclusions]Sijicao granules could play an important role for the treatment of eczema through multi-component,multi-target,multi-pathway and their interaction.

The Stimulatory Effects of the Antimicrobial Agents Bavistin, Cefotaxime and Kanamycin on <i>In Vitro</i>Plant Regeneration of <i>Centella asiatica</i>(L.)—An Important Antijaundice Medicinal Plant

Antimicrobial agents such as bavistin, cefotaxime and kanamycin were evaluated for their effects on the rapid shoot regeneration from nodal explants of Centella asiatica (L.). Filter sterilized bavistin (250 mg/L) was augmented alone and in combination with cytokinins such as BAP and TDZ into the media to trace the effect on regeneration. On this basis, the potential use of bavistin (150 mg/L) along with BAP (2.0 mg/L) was evaluated which showed the maximum shoot number (6.6) and shoot length (4.4 cm) respectively. Cefotaxime at the concentration of 100 μM/L was found to be effective to obtain the maximum shoot number formation (5.8) with the regeneration frequency (90%). Kanamycin at the concentration of 80 μM/L induced maximum shoot regeneration (5.12). Kanamycin at 100 μM/L or at higher concentrations reduced the shoot regeneration. The best rooting response was noticed when in vitro regenerated microshoots were transferred to the rooting medim which was supplemented with IBA (2.0 mg/L). This combination generates 90% of regeneration frequency and maximum number of roots per shoot (14.2) and high root length (4.2 cm). The rooted plants were acclimatized and transferred to field for survivalance. The addition of antibiotics was found to be more effective and safer for using since their effects on regeneration were found to be negligible.

HPLC-ELSD法测定积雪草中积雪草苷及羟基积雪草苷的含量

积雪草提取物通常被用作化妆品原料,有美白、抗氧化等功效,发挥此功效的主要物质有积雪草苷、羟基积雪草苷。采用高效液相色谱-蒸发光散射检测(HPLC-ELSD)技术,建立了测定积雪草中积雪草苷和羟基积雪草苷含量的方法。选用Unitary C18色谱柱(4.6×250 mm,5μm),以乙腈-水为流动相,梯度洗脱,流速1 mL/min,柱温为30℃,采用Agilent 1260 InfinityⅡ蒸发光散射检测器(漂移管温度70℃,载气流速1.6 L/min)。研究表明,积雪草苷和羟基积雪草苷质量浓度分别在100~702 mg/L,105~738 mg/L范围内,峰面积与质量浓度线性关系良好,相关系数分别为0.9996和0.9997。积雪草苷和羟基积雪草苷平均加样回收率分别为101.20%,98.30%。相对标准偏差(RSD)均在3%以内。方法简单,准确度高,分离效果好,干扰少,可用于积雪草中积雪草苷和羟基积雪草苷的含量测定。

Antifertility properties ofCentella asiatica ethanolic extract as a contraceptive agent: Preliminary study of sperm proteomic

Objective:To investigate the antifertility properties ofCentella asiatica L. ethanolic extract and identify the sperm proteomic changes in treated male rats.Methods: A total of 32 male Sprague-Dawley rats were divided into two groups: control group (n=16) received distilled water and treatment group (n=16) which received 300 mg/kg ofCentella asiatica L. extract for 42 d. After the treatment period, the number of implantation sites was recorded and the sperm proteomic changes were analysed by 2D gel electrophoresis. In addition, the expression of protein spots was quantified by MALDI-TOF analysis.Results:Centella asiatica L. extract resulted in low number of implantation sites in the treatment group (100.00±2.82) compared to the control group (183.00±2.14). The percentage of infertile male rats in the treatment group was higher (43.75%) compared to the control group (18.75%). Proteomic analysis showed the expression of protein spots identified in the treatment group decreased with 234 spots compared to the control group with 282 spots.Conclusions:The results revealed that there was antifertility activity in the male rats with the administration ofCentella asiatica L. ethanol extract and the identified proteins could provide understanding on the adverse effect ofCentella asiaticain male reproductive system.

Vegetative Growth and Molecular Identification of Fusarium equiseti Isolated from Wilt Disease of Centella asiatica L. in Bangladesh

Centella asiatica (L.) is commonly known as Thankuni plant and has ethnobotanical importance in Bangladesh. Present experiment was conceded to investigate the wilt disease of C. asiatica, vegetative growth and molecular characterization of pathogenic fungi. Pathogenic fungus, Fusarium equiseti was identified as a causal agent of wilt disease in C. asiatica. The effect of culture media on the mycelial growth of F. equiseti showed the highest (89.25 mm) on potato dextrose agar (PDA) medium followed by carrot agar (CA) medium and the lowest growth (40.25 mm) was measured in HA medium. The optimal temperature and pH for mycelial growth of F. equiseti were 30&deg;C and 7, respectively. The genetic variation of the selected species of fungi, the internal transcribed spacer (ITS) region was amplified using ITS4 and ITS5 primers and sequenced. The PCR product of the ITS region of F. equiseti was 535 bp. Phylogenetic tree of thirty-seven strains of Fusarium sp. based on the nucleotide sequences of the ITS region using the neighbor-joining method with 1000 bootstrapping indicated that 98% - 100% identity with MN886590.1 JUF0046 (F. equiseti). ITS sequences are generally constant, or show little variation within species, but vary between species in a genus. The ITS region is relatively short and can be easily amplified by PCR using universal single primer pairs. Genetic distance exhibited high level of similarity with identical ITS sequences. To date, no published research articles are found on the molecular identification of F. equiseti, the causal agent of fusarium wilt disease of C. asiatica in Bangladesh.

Induced Mutation for Developing Mutant Rice Lines Tolerant to the Parasitic Weed Striga asiatica (L.) Kuntze

This work aims to screen mutant rice lines tolerant to Striga asiatica.Two rainfed sensitive rice varieties B22 and F154 were used.Plants survival rates of the two parents were significantly lower respectively(9.74a and 11.83a)than those of mutant lines(55.36c to 74.36b);Striga plants emergence/pot were significantly higher for the parents(13.96c and14.89c)compared to the mutants(0.12a to 1.5b);the infection rate of parents(7.37b;7.86b)was higher compared to the mutants(2.27a to 2.74a);fertility rate/plant of parents was lower(20.98%b;22.29%b)but much higher than mutants(72.19%b to 78.35%b);the average panicle number/plant of parents was significantly lower(0.5a;1a)than those of mutants(1.5b to 2.4bc)and the 100 g grain weight of parents are lower(2.35a;2.56a)than those of mutants(3.19b to 3.23b).The culture of those mutant lines may increase rice production and contribute to enhancing food security in Madagascar.

A new triterpene and a saponin from Centella asiatica

A new triterpene and a saponin, named 2α,3β23-trihydroxyurs-20-en-28-oic acid （1） and 2α,3β,23-trihydroxyurs-20-en-28-oic acid O-α-L-rhanmopyranosyl-（1→4）-O-β-D-glucopyranosyl-（1→6）-O-β-D-glucopyranosyl ester （2）, have been isolated from the aerial part of CenteUa asiatica. Their structures were elucidated by spectral methods, including 2D-NMR spectra.

Determining the effect of Plantago major L. and Rosa damascene herbal ointment for bedsore healing

Background:Bedsores are damage or loss of skin integrity due to prolonged pressure which there are several ways to treat and prevent them.The aim of the study was to determine the effect of Plantago major L.and Rosa damascene herbal ointment for bedsore healing in hospitalized patients.Methods:This clinical trial study was accomplished with 100 patients divided into control(50 patients)and intervention(50 patients)groups.The control group had received routine treatment(dressing with Comfeel),but the intervention group only had received herbal ointment and then both groups were compared with each other through the pressure ulcer scale for healing(PUSH)tool.Results:The mean and standard deviation of recovery time in the Comfeel group was 27.40±12.467 and in the herbal ointment group was 21.57±12.695.In the pre-intervention evaluation,the mean and standard deviation of the PUSH score in the herbal ointment group was 10.44±0.323 while in the last evaluation in the same group it was 0.68±0.288 but in the Comfeel group,in the pre-intervention evaluation,the mean and standard deviation of the PUSH score was 10.68±0.317,while in the last evaluation it was 1.98±0.472.The highest frequency of bedsores in the study units was in the sacral region(49%)and the lowest frequency was in the sole of the foot with 1%.Conclusion:The results showed that Plantago major L.and Rosa damascene herbal ointment was effective in accelerating recovery time and wound healing.

大粒车前子多糖组分PLP-1的结构表征

研究了由大粒车前子多糖分离纯化所得组分PLP-1的一级精细结构特征.通过单糖组成分析、部分酸水解及甲基化分析,并结合NMR,GC和GC-MS等技术测定了PLP-1的一级结构.结果表明,PLP-1由Ara(33.98%),Xyl(58.23%),Rha(2.36%),Glc(2.23%),Gal(2.36%)和Man(0.83%)组成,糖醛酸以GlcA形式存在.PLP-1中主要糖残基有α-T-linked Araf(9.58%),β-T-linked Xylp(8.71%),α-1,3-linked Araf(18.22%),β-1,3-linked Xylp(8.05%),β-1,4-linked Xylp(5.85%),β-1,2,4-linked Xylp(16.98%)和β-1,3,4-linked Xylp(27.52%),GlcA以α-T-linked GlcAp形式存在;此外还有1,2-linked Rhap,T-linked Glcp,1,6-linked Glcp,T-linked Galp,1,3-linked Galp和1,3,6-linked Galp等少量其它残基.根据所得结果推断出PLP-1可能的一级结构.

中药车前子对小鼠气囊滑膜炎细胞因子TNF-α及IL-12影响的实验研究

目的:研究中药车前子对小鼠滑膜炎的抗炎机制。方法:使用T.Mikami's方法用角叉菜胶给小鼠造模,应用双抗体夹心酶联免疫吸附试验法检测灌洗液中TNF-α、IL-12的含量变化。结果:中药车前子组TNF-α、IL-12含量低于对照组,有显著性差异(P<0.01)。结论:车前子能通过抑制滑膜炎症中TNF-α,IL-12的含量进行抗炎。

车前草中熊果酸、齐墩果酸的HPLC测定

采用 ＨＰＬＣ法同时测定车前草中熊果酸、齐墩果酸的含量。选用 Ｃ_１８ ＯＤＳ色谱柱，流动相：甲醇－水（８８： １２），检测波长：２２０ ｕｍ，流速：０． ６ ｍＬ／ｍｉｎ，柱温：２５ ℃。熊果酸和齐墩果酸分别在 ０．４０４２～２．０２０ μｇ（ｒ＝０． ９９８４）和 ０．４１０～２．０５０ μｇ（ｒ＝０．９９９７）范围内呈线性；熊果酸、齐墩果酸的平均回收率分别为９８．２％（ＲＳＤ＝１．７４％）和９７．８％（ＲＳＤ＝２．２４％）。该法准确、快速、灵敏，重现性好。

HPLC法快速测定车前草中熊果酸的含量

改进了高效液相色谱法(HPLC)测定熊果酸的方法,得出最佳色谱条件:AtlantisTMC18分析柱(4.6mm×250mm,5μm);流动相为0.02mol/L磷酸二氢钠∶甲醇(体积比为15∶85);检测波长为215nm;流动相流速为0.8mL/min。线性范围:59～950μg/mL,回归方程为Y=201.55X-2070.9,r=0.9997。在上述条件下对中药车前草中所含熊果酸进行了含量测定,分析结果表明,此法简单快速、高效灵敏、重现性好。