N-alkylamide profiling of Achillea ptarmica and Achillea millefolium extracts by liquid and gas chromatography–mass spectrometry

Achillea millefolium and Achillea ptarmica are both plants belonging to the Asteracea family and are traditionally used for their medicinal properties. It has already been shown that some N-alkylamides(NAAs)are responsible for these pharmacological actions. Therefore, in the present study, the NAA content of the two plants was analytically characterised. Different extracts were prepared from the roots, the leaves, the stems and the flowers. The structures of NAAs have been assigned in ethanolic extracts of Achillea millefolium and Achillea ptarmica using high performance liquid chromatography – electrospray ionisation – mass spectrometry(HPLC–ESI–MS) and gas chromatography – electron impact – mass spectrometry(GC–EI–MS). Using both analytical techniques, the structures of 14 and 15 NAAs have been assigned in Achillea ptarmica and Achillea millefolium, respectively. Structures of two new NAAs, previously never observed in Achillea ptarmica,were assigned: deca-2E,6Z,8E-trienoic acid 2-methylbutylamide(homospilanthol) or a related isomeric compound and deca-2E,4E-dienoic acid N-methyl isobutylamide. The structure of homospilanthol or a related isomeric compound was also assigned in Achillea millefolium for the first time.

Biochemical,Enzymatic,and Immunological Study on Antimutagenic Achillea millefolium Methanolic Extract in vivo

Traditionally medicinal plants are used for its potential chemotherapeutic action and for safety upon the continued use of these plants.Achillea millefolium(AM)is an ancient herb which is used to treat wounds,headaches,disorder in gastrointestinal and hepatobiliary,pain,and inflammation.This study was aimed to investigate the biochemical parameter of antimutagenic methanolic extract for AM in vivo.Plant extract together with MTX decreased the damage caused by the drug on the concentrations of total cholesterol and triglycerides(160.3±2.26 and 108.7±2.71),significant increase in total protein concentration(8.51±0.17,9.46±0.19 and 10.8±2.71 g\dL)for 100 and 200 mg\kg was observed.Also plant extracts counteract the damage caused by drug through the reduction for both creatinin and albumin concentrations to 1.57±0.03 mg\dL and 4.56±0.09 g\dL,respectively.On enzymatic level,the results of interactions indicated the ability of plant to modulating harmful effects of the drug(57.1±1.3,130±3.1 and 111.1±2.1 U\L)for GOT,GPT and ALP,respectively.On the other hand,the plant possesses the ability to enhance the concentrations of all immunoglobulin(IgM,IgA and IgG)in the two doses tested.Therefore,the results of this study indicate the antimutagenic potential of AM extract and encouraging its consumption with MTX as one of the promising therapies.

Assessments of Immunological Activity of Achillea Millefolium Methanolic Extract on Albino Male Mice

Achillea millefolium （Asteraceae） is a permanent herb highly recognized in traditional medicine for its anti-oxidant and anti-inflammation properties. However, studies on phytochemical constituents of A. millefolium underlying these properties are scarce. The present work focuses on examining the effect of methanol extract of A. millefolium L. on total and differential blood cells account on albino male mice. The results showed the methanol extract increased the account of lymphocyte, and monocyte cells, and total account as well as this extract showed high decrease in the oxidative stress of MTX after the interfere between the extract and MTX due to increase in the leucocyte cells compared with controls. Concluded from these results that methanol extract of A. millefolium has ability enhancement in leucocyte cells in the blood and it has detoxification effect of MTX.

Achillea fragrantissima,rich in flavonoids and tannins,potentiates the activity of diminazine aceturate against Trypanosoma evansi in rats

Objective:To evaluate activity of methanol extract of Achillea fragrantissima(meth)(A.fragrantissima) alone or in combination with diminazine aceturate(DA) against Trypanosoma evansi in experimentally infected rats.Methods:Sixty adult male Wister albino rats were divided equally into 6 groups(A-F).Rats in groups A-E were experimentally infected with T.evansi and those in group F were uninfected.The groups were treated respectively as follows:group A- with 3.5 mg/kg DA;group B- with 1 000 mg/kg meth,A.fragrantissima;group C-3.5mg/kg DA plus 500 mg/kg meth A.fragrantissima;group D-3.5 mg/kg DA plus 1 000 mg/kg meth A.fragrantissima.Group E was left untreated.Parasitaemia,survivability,packed cell volume,hemoglobin concentration,total leucocytes count,lymphocyte count,and serum malondialdehyde and reduced glutathione(GSH) levels were estimated.Phytochemical screening of meth A.fragrantissima was also performed.Results:The phytochemical analysis of the meth A.fragrantissima indicated a higher content from polyphenols tannins and non tannins and flavonoids.The efficacy percentage against trypanosomiasis in groups A to E was respectively as follows 80,40,90.100,0.The administration of meth-A.fragrantissima(1000)mg/kg b.wt.) produced a moderate efficacy against trypanosomiasis.Untreated rats in group E died between 25 and 30 d post infection.The rats given DA and meth A.fragrantissima combinations(C and D) showed faster and higher recovery rates than the uninfected control and groups A and B.The initial reduction in packed cell volume,hemoglobin,total leucocytes count,increases in serum malondialdehyde and decreases in GSH levels were reversed by the treatments.C onclusions:The administration of the methanol extracts of A.fragrantissima and DA combination therapy was more effective than each product alone in the treatment of rats infected with Trypanosoma evansi and further studies are required to isolate more active ingredients.

Vegetation Analysis and Soil Characteristics on Two Species of Genus <i>Achillea</i>Growing in Egyptian Desert

The present study provides a vegetation analysis and species distribution at 50 sites, emphasizing the environmental factors that affect species distribution. A total of 74 plant species belonging to 67 genera and related to 23 families of vascular plants are recorded. Asteraceae, Poaceae, Chenopodiaceae, Brassicaceae, Fabaceae and Zygophyllaceae are the largest families, and therophytes (41.89) and chamaephytes (24.32%) are the most frequent, indicating a typical desert life-form spectrum. Chorological analysis revealed that 25 of the studied species were Mediterranean taxa, Saharo-Sindian chorotypes, either pure or penetrated into other regions, comprised 47 species. After application of the TWINSPAN and DCA programs, 4 vegetation groups (A-D) were identified, groups A and B were dominated by Achillea santolina, group C was codominated by Zygophyllm coccinum and Launaea spinosa and group D was dominated by Leptadenia pyrotechnica. Groups A and B may represent the vegetation types of the Western Mediterranean coast of Egypt, while groups C and D may represent the Wadi Hagul. The linear correlation of soil variables with the importance values of some dominant species and the application of Canonical Correspondence Analysis (CCA-biplot) indicates significant associations between the floristic composition of the studied area and the edaphic factors such as electrical conductivity, pH, calcium carbonate, sulphates, bicarbonate, cations (Na+, K+, Ca++ and Mg++) and PAR.

Biological activities of the hydro-alcoholic and aqueous extracts of <i>Achillea fragrantissima</i>(Forssk.) grown in Jordan

Aqueous and hydro-alcoholic extracts of Achillea fragrantissima L. (Asteraceae) grown in Jordan were screened for their antioxidant, antimicrobial, antiplatelet, anti-proliferative and acetylcholinesterase (AChE) inhibition efficacy. Total phenols and flavonoids were determined colorimetrically. The radical scavenging activities were evaluated using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2’-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical scavenging activity assays. High performance liquid chromatography-mass spectrometry (HPLCMS) analysis resulted in the identification of 7 phenolic compounds in the hydro-alcoholic extract and 4 compounds in the aqueous extract;quercetin 3-β-D-glucoside was the main component for both extracts. Antimicrobial activities were determined by antimicrobial susceptibility testing such as agar well-diffusion method, minimum inhibition concentration and minimum bactericidal concentration. Gram positive bacteria showed sensitivity to hydro-alcoholic extract in the agar-well diffusion test. No significant activity was observed against gram negative bacteria and Candida albicans. Hydro-alcoholic extract had a bactericidal activity against Streptococcus pneumoniae and Bacillus cereus at high concentrations (MIC 12.5 mg/ml) rather than inhibitory effect. In vitro antiplatelet activity was tested on human whole blood using an electrical impedance method. At concentrations (50, 100, and 200 μg/ml), no effect on platelet aggregation was noticed. Anti-proliferative activity was investigated using the MTT assay. At concentrations up to 200 μg/ml, extracts did not possess cytotoxic activity against the MCF-7 cells. Acetylcholinesterase (AChE) inhibitory capacity of A. fragrantissima extracts was tested using TLC assay method, and neither aqueous, nor hydroalcoholic extracts showed AChE inhibition. The present investigation supported the traditional use of A. fragrantissima in the Jordanian folk medicine as an antimicrobial active representative of the genus Achillea. A. fragrantissima extracts should be further studied for their potential use in preventing/treating diseases in which oxidative stress is a part of the pathophysiology.

Allelopathic Effect of Achillea （Achillea santolina） on Germination and Growth of Crop Plants

In order to investigate the likely allelopathic potential of an aqueous extract and powder of Achillea （,4chillea santolina） on the germination and seedling growth of pea （Cicer arietinum）, safflower （Carthamus tinctorius L.） and wheat （Triticum sativum）, an experiment was carried out at the laboratory and greenhouse of agriculture faculty of Sho3hid Bahonar University of Kerman, Iran, from October 2009 to March 2010. A completely randomized design with three replicates using five extract concentrations of achillea plant including 0, 25, 50, 75, and 100 g Ll and four amounts of powder of it including 0, 7.5, 15 and 30 g kg1 of soil was employed in laboratory and greenhouse experiments, respectively. All extract concentrations of achillea except 25 g Ll inhibited pea and wheat seed germination significantly, but had no inhibitory effect on the germination of safflower, The powder of mature achillea plants affected the fresh and dry weight and shoots length in these crops negatively compared with the control in all levels. Therefore use of this plant should be prevented in rotation or intercropping with these three crop plants. Further research conducted in the analytical laboratory as well as in the field is needed before a practical application of the extract and powder as weed inhibiting agent can be recommended.

Evaluation of dietary addition of 2 essential oils from Achillea moschata,or their components (bornyl acetate,camphor,and eucalyptol) on in vitro ruminal fermentation and microbial community composition

This study investigated the effects of 2 Achillea moschata essential oils extracted from plants collected in 2 different valleys of the Italian Alps and 3 pure compounds of oils—bornyl acetate(BOR),camphor(CAM),and eucalyptol(EUCA)—on in vitro ruminal fermentation and microbiota.An in vitro batch fermentation experiment(Exp.1)tested the addition of all of the substances(2 essential oils and 3 compounds)in fermentation bottles(120 mL)at 48 h of incubation,whereas a subsequent in vitro continuous culture experiment(Exp.2)evaluated the pure compounds added to the fermenters(2 L)for a longer incubation period(9 d).In both experiments,total mixed rations were incubated with the additives,and samples without additives were included as the control(CTR).Each treatment was tested in duplicate and was repeated in 3 and 2 fermentation runs in Exp.1 and 2,respectively.Gas production(GP)in Exp.1 was similar for all of the treatments,and short chain volatile fatty acid(SCFA)production was similar in both experiments except for a decrease of SCFA produced(P=0.029)due to EUCA addition in Exp.2.Compared to CTR,BOR and CAM reduced the valerate proportion(P=0.04)in Exp.1,and increased(P<0.01)the acetate proportion in Exp.2.All treatments increased(P<0.01)total protozoa counts(+36.7%and+48.4%compared to CTR on average for Exp.1 and 2,respectively).In Exp.1,all of the treatments lowered the Bacteroidetes and Firmicutes and increased the Proteobacteria relative abundances(P<0.05),whereas in Exp.2,the EUCA addition increased(P=0.012)the Ruminococcus.In Exp.1,methane(CH4)as a proportion of the GP was lowered(P=0.004)by the addition of CAM and EUCA compared to CTR,whereas in Exp.2,EUCA reduced the amount of stoichiometrically calculated CH4 compared to CTR.Overall,essential oils extracted from A.moschata and the pure compounds did not depress in vitro rumen fermentation,except for EUCA in Exp.2.In both experiments,an increase of the protozoal population occurred for all the additives.

2,3-Seco and 3-nor guaianolides from Achillea alpina with antidiabetic activity

In this study,we presented the isolation and characterization of eight novel seco-guaianolide sesquiterpenoids(1-8)and two known guaianolide derivatives(9 and 10),from the aerial part of Achillea alpina L..Compounds 1-3 were identified as guaianolides bearing an oxygen insertion at the 2,3 position,while compounds 4-8 belonged to a group of special 3-nor guaianolide sesquiterpenoids.The structural elucidation of 1-8,including their absolute configurations,were accomplished by a combination of spectroscopic data analysis and quantum electronic circular dichroism(ECD)calculations.To evaluate the potential antidiabetic activity of compounds 1-10,we investigated their effects on glucose consumption in palmitic acid(PA)-mediated HepG2-insulin resistance(IR)cells.Among the tested compounds,compound 7 demonstrated the most pronounced ability to reverse IR.Moreover,a mechanistic investigation revealed that compound 7 exerted its antidiabetic effect by reducing the production of the pro-inflammatory cytokine IL-1β,which was achieved through the suppression of the NLRP3 pathway.

基于指纹图谱、多指标定量和网络药理学的蓍草质量标志物预测分析

目的:采用指纹图谱和网络药理学的方法,分析预测蓍草的潜在中药质量标志物(Q-Marker)。方法:采用高效液相色谱法建立蓍草的指纹图谱,对21批蓍草进行相似度评价并对共有峰进行指认和归属,再运用网络药理学方法,构建“成分-靶点-通路”网络图,分析预测蓍草的Q-Marker,并测定其含量。结果:建立了21批蓍草的指纹图谱,相似度均大于0.910,确认了13个共有峰,通过对照品比对指认出6个色谱峰并测定其含量,6个峰分别为绿原酸、异绿原酸B、异绿原酸A、异绿原酸C、百蕊草素Ⅱ和3,4,5-三咖啡酰奎宁酸。经网络药理学分析筛选出发挥抗炎、抗菌作用的绿原酸、百蕊草素Ⅱ和3,4,5-三咖啡酰奎宁酸3个活性成分,38个核心靶点和20条关键通路。初步预测绿原酸、百蕊草素Ⅱ和3,4,5-三咖啡酰奎宁酸可作为蓍草潜在的Q-Marker。结论:预测分析得到的蓍草QMarker,为蓍草药材质量控制提供一定参考,为后续其药效物质基础及作用机制研究奠定基础。

云南蓍地上部分化学成分分离与结构鉴定

针对药用植物云南蓍的化学成分进行分离与结构鉴定,采用正相硅胶、反相硅胶(Rp-C18)、葡聚糖凝胶(Sephadex LH-20)、MCI以及RP-HPLC色谱等几种分离手段,对该药用植物地上部分的90%乙醇提取浸膏进行分离纯化,依据现代波谱方法(核磁共振NMR、质谱MS)以及理化性质对分离得到的单体化合物的结构进行确定。最终从该植物中分离并鉴定了9个单体化合物,结构分别为matricarin(1)、3β-hydroxy-11α,13-dihydro-costunolide(2)、黑麦草内酯(3)、4-羟基-3-甲氧基苯乙酮(4)、2-苯基乙酰胺(5)、sylvamide(6)、5-羟甲基呋喃甲醛(7)、6-甲氧基-7-羟基香豆素(8)、邻苯二甲酸二丁酯(9)。这些化合物中,化合物1~8为首次从该植物中分离得到。

蓍草组织培养的研究

以MS和 8114为基本培养基 ,通过调整激素种类与浓度等培养条件 ,按正交实验设计的原则 ,摸索出建立蓍草组织培养体系的最佳条件 :愈伤诱导最佳培养基为 8114+6BA1.0 +NAA1.0 +ZT1.0 ;最佳分化培养基为 8114+6BA4.0 +NAA0 .2 +ZT1.0 ;最佳生根培养基是 1/2MS0 +0 .1NAA .

菊科蓍属10种植物花粉的形态学研究

目的 :研究 10种蓍属Achillea植物的花粉形态。方法 :对 10种植物的花粉进行了光镜 (LM)和扫描电镜 (SEM)的观察。结果 :蓍属 10种植物的花粉均为近球形 ,萌发孔为三孔沟 ,表面具瘤状 刺状 穴状纹饰。但在花粉大小、萌发沟形态和表面纹饰等方面仍有区别。结论 :蓍属 10种植物的花粉在形态上存在细微的种间差异。

蓍草提取物预防慢性肝损伤、肝纤维化的实验研究

背景与目的:研究蓍草提取物对大鼠实验性慢性肝损伤、肝纤维化的预防作用,并探讨其作用机制。材料与方法:采用40%CCl4皮下注射制备肝纤维化大鼠模型,分别用3种不同浓度的蓍草提取物进行干预,观察检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)和肝组织病理学变化。结果:蓍草提取物预防给药能降低CCl4所致大鼠血清中异常增高的ALT、AST含量,光镜下观察肝纤维化模型组大鼠肝细胞严重变性、坏死,胶原纤维明显增加,蓍草预防各组大鼠肝细胞病理损伤得到明显改善。结论:蓍草提取物对大鼠实验性肝损伤具有一定程度的预防作用和抗肝纤维化作用。

牛胆粉等3种特色蒙药材的薄层色谱鉴别研究

目的建立特色蒙药材牛胆粉、蓍草、酸梨的薄层鉴别标准。方法采用薄层色谱法对牛胆粉、羊胆粉、蓍草、酸梨进行定性鉴别。结果薄层分离度好,斑点清晰,重现性良好。结论该方法简便、可靠、准确,可用于蒙药牛胆粉、蓍草、酸梨的质量控制。

蓍草总倍半萜内酯对类风湿性关节炎模型大鼠保护作用的实验研究

目的探讨蓍草总倍半萜内酯对类风湿性关节炎(RA)模型大鼠的抗炎作用,并初步探讨其作用机制。方法大鼠右后足跖皮下注射弗氏完全佐剂建立佐剂性关节炎大鼠模型,给予不同剂量蓍草总倍半萜内酯和雷公藤多苷药物干预21d。通过检测大鼠关节肿胀度、关节炎指数和血清白介素(IL)-6、IL-1β、肿瘤坏死因子(TNF)-α水平等指标,观察蓍草总倍半萜内酯片对佐剂性RA大鼠的影响。结果与模型组比较,蓍草总倍半萜内酯中、高剂量组在14、21、28d大鼠足跖肿胀度显著低于模型组(P<0.05,P<0.01),明显抑制了RA大鼠足关节肿胀度。蓍草总倍半萜内酯中、高剂量组在14、21、28d大鼠关节指数显著低于模型组(P<0.05,P<0.01)。与模型组比较,蓍草总倍半萜内酯低、中、高剂量组均能够显著降低IL-6、IL-1β、TNF-α水平(P<0.05,P<0.01),各剂量组的抑制作用呈剂量依赖性。结论蓍草总倍半萜内酯具有较好的治疗大鼠RA作用,其作用机制可能与下调炎症因子IL-6、IL-1β、TNF-α水平有关。

3种宿根花卉花期控制技术的研究

通过增加越冬保护、施用植物生长调节剂的方法,提早千叶蓍、桔梗的花期。其中温室盆栽能使千叶蓍、桔梗花期提早到"五一"期间,千叶蓍花期提早23 d,桔梗花期提早54 d,效果显著。对假龙头进行延迟花期的实验研究,其中通过延迟播种能延迟花期48 d;通过品种筛选,品种间自然花期相差40 d;通过不同时期重压剪,能延迟花期40～100 d;通过适时剪除花穗,能促进假龙头进入第二次花期,有效地将花期控制到预期的时期;通过叶面喷施B9,能延迟花期5～7 d。

新疆千叶蓍抑菌作用研究

[目的]研究不同方法提取的千叶蓍提取物的抑菌活性,并且测定千叶蓍提取物的最低抑菌浓度和最低杀菌浓度,为千叶蓍的开发利用提供依据。[方法]分别用热提取法和冷浸法提取千叶蓍的有效成分,以大肠杆菌、青霉和部分病原菌等作为供试菌种,采用Bauer-Kirby法(滤纸片法和微量2倍连续梯度稀释法试验)对几种常见菌株进行体外抑菌试验。[结果]千叶蓍的浓度85%乙醇、浓度95%乙醇、石油醚和乙酸乙酯提取物对大肠杆菌、金黄色葡萄球菌、变形杆菌和灰霉等都有抑菌效果。浓度85%乙醇提取物对大肠杆菌的MIC为0.031 25 g/ml,MBC为0.062 50 g/ml;对金黄色葡萄球菌的MIC为0.062 50 g/ml,MBC为0.125 00 g/ml;对变形杆菌的MIC为0.062 50 g/ml,MBC为0.125 00 g/ml。[结论]不同方法提取的千叶蓍提取物均有一定的抑菌作用。

蓍草提取物防治肝纤维化大鼠脂质过氧化作用的实验研究

为研究中药蓍草对肝组织纤维化过程中脂质过氧化的防治作用,观察了蓍草醇提取物防治CCL4诱发的大鼠肝纤维化过程中脂质过氧化作用的效果。结果:蓍草醇提物可有效提高肝组织超氧化物岐化酶(SOD)的含量、降低自由基脂质过氧化反应终产物丙二醛(MDA)的含量。结论:蓍草醇提取物可能通过减轻氧自由基对肝细胞的破坏对肝细胞起到保护作用。

基于网络药理学和UPLC-Q-TOF/MS结合动物实验探究蓍草预防急性肝损伤的作用机制

目的 通过UPLC-Q-TOF/MS定性分析和网络药理学预测并结合动物实验验证初步探究蓍草保护急性肝损伤(ALI)大鼠肝脏的作用机制。方法 采用UPLC-Q-TOF/MS法鉴定蓍草中的化学成分,利用TCMSP、CTD和Similarity Ensemble Approach等数据库筛选这些成分的作用靶点,同时在GeneCards、OMIM和NCBI数据库筛选ALI相关靶点,将成分作用靶点与疾病靶点的交集靶点导入STRING构建蛋白质-蛋白质相互作用(PPI)网络,并使用Cytoscape3.8.2软件进行网络拓扑分析筛选核心靶点,通过DAVID数据库对关键靶点进行GO和KEGG富集分析。通过肝组织HE染色,大鼠血清中肝功能指标、炎症因子水平以及肝组织中氧化应激水平的测定对蓍草防治ALI的药效进行评价,并利用免疫组化的方法对预测的核心靶点及KEGG富集通路结果进行验证。结果 经UPLC-Q-TOF/MS法鉴定出20个成分,网络药理学方法筛选得到上述成分预防治疗ALI的核心靶点104个,GO功能主要与炎症反应、氧化还原酶活性和蛋白酶结合等生物过程相关,KEGG结果主要涉及IL-17、TNF及Toll样受体等信号通路;动物实验结果显示,蓍草预防性给药后能够缓解大鼠肝脏的损伤,显著降低模型组大鼠血清中ALT、AST、TNF-α、IL-6、IL-1β水平和肝组织中MDA的含量,且SOD和CAT在肝组织中的活性显著增强;此外还发现蓍草给药后可上调大鼠肝脏组织中Nfr2蛋白的表达,下调TLR-4、NF-κBp65、keap1和HO-1蛋白的表达。结论 蓍草对四氯化碳诱导的ALI大鼠肝脏具有保护作用,该作用可能与其调节keap1/Nrf2/HO-1和Toll样受体信号通路有关。