Methanolic extract of Ephedra alata inhibits breast cancer cells in vitro and in vivo

Objective:To determine the anticancer potential of the methanolic extract from Ephedra alata against breast cancer both in vitro and in vivo.Methods:The effects of the methanolic extract of Ephedra alata on the viability,migration as well as apoptosis of breast cancer 4T1 cells were measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay,Transwell assay,and annexin V-FITC staining assay,respectively.Histological examination was also carried out.Moreover,a murine breast cancer model was established to evaluate the inhibitory effect of the extract.Biochemical parameters including hepatic and non-hepatic enzymes,malondialdehyde,and glutathione were investigated.Results:The methanolic extract of Ephedra alata showed a strong anti-proliferative and anti-migratory activity against 4T1 cells in a dose-dependent manner.It also induced apoptosis in 4T1 cells.In an in vivo mouse model,the extract markedly inhibited tumor growth,reduced malondialdehyde,and hepatic and non-hepatic enzymes as well as increased glutathione level.Conclusions:The methanolic extract of Ephedra alata inhibits breast cancer in vitro and in vivo,which may be a promising anticancer agent.

Preparation of Ephedra houttuynia Granule and Its Therapeutic Mechanism of Anti-Mycoplasma gallisepticum Infection Based on Network Pharmacology

The use of Chinese herbal medicines can replace antibiotics that cause drug-resistance problems,which are currently necessary for disease control.In this paper,a traditional Chinese medicine compound named Ephedra houttuynia granule for the treatment of Mycoplasma galliscepticum(MG)infection was prepared.Furthermore,its action mechanism was explored through network pharmacology.The optimal extraction and granulation processes of the compound were determined by high performance liquid chromatography(HPLC)method and L9 orthogonal test,and in the treatment experiment,Ephedra houttuynia granule showed a significant therapeutic effect on MG infection.In the study of network pharmacology,the results showed that the core targets of Ephedra houttuynia granule against MG infection were vascular endothelial growth factor(VEGFA),fos proto-oncogene(FOS),prepro-coagulation factor II(F2),etc.,the gene ontology/kyoto encyclopedia of genes and genomes(GO/KEGG)analysis results indicated that the signaling pathways of neuroactive ligand receptor interaction,cAMP,IL-17,T cell receptor,and tumor necrosis factor(TNF)might involve in anti-MG infection.In conclusion,this study would provide a new idea for elucidating the action mechanism of other diseases in veterinary clinic,which had a certain guiding significance.

新疆洋海古代麻黄的化学成分研究

[目的]通过对古代麻黄中化学成分进行分离鉴定来探讨其化学成分的稳定性。[方法]采用乙醇回流提取法提取麻黄中的化学成分,并采用色谱分离技术进行分离纯化,然后通过波谱分析鉴定其化合物的结构。[结果]从古代麻黄乙醇提取物中分离鉴定了8个化合物,分别为山柰酚-3-O-α-L-吡喃鼠李糖苷(1)、槲皮素-3-O-β-L-吡喃鼠李糖苷(2)、山柰酚-3-O-α-L-吡喃鼠李糖苷-4″-E-(4-羟基)-肉桂酸酯(3)、刺五加酮(4)、胆甾-5-烯-3β-正十四酸酯(5)、β-谷甾醇(6)、正二十八烷醇(7)和正二十六羧酸(8)。[结论]研究发现,麻黄中黄酮类成分结构比较稳定。

历代麻黄临床用量评述

本文采用文献研究与数据统计的方法,将中药麻黄自先秦至今两千年的临床剂量变化进行了统计和描绘。结果表明:在金元时期之前麻黄用量较大,而从金元始,麻黄剂量明显下降。通过此文对麻黄确凿剂量的核算及大剂量应用的回顾,或可对拓宽麻黄的运用思路有所裨益。

提取麻黄多糖的2种方法比较

[目的]比较热水浸提法和微波辅助法提取麻黄多糖的工艺和效果。[方法]采用热水浸提法和微波辅助法对麻黄中的多糖进行提取,用正交试验优选了热水浸提法和微波辅助法提取麻黄多糖的最佳工艺条件。[结果]热水浸提法提取的最佳工艺条件为:提取温度60℃、提取时间3 h、料液比1∶10(g/ml);在此条件下,麻黄多糖的提取率为6.31%;微波辅助法提取的最佳工艺条件为:微波功率420W、微波处理时间60 s、料液比1∶10(g/ml);在此条件下,麻黄多糖的提取率为5.57%。[结论]热水浸提法提取效率高、条件温和易控制,是理想的麻黄多糖提取方法。

Studies on a Traditional Chinese Medical Prescription Ma Xing Shi Gan Tang by Bio-coordination Chemistry

本文报道 pH 电位滴定法测出麻黄碱与生石膏中的 Cu^(2+)、Fe^(2+)、Zn^(2+)、Mn^(2+)离子形成的 ML 及 ML_2型络合物的稳定常数。滴定曲线显示甘草酸与上述四种金属离子也能分别形成不同稳定程度的络合物。用麻黄碱加甘草酸和上述四种金属离子配制成的模拟麻杏石甘汤和单纯生石膏煎剂与麻性石甘汤一样均有使家兔退热的效果,与对照组比较均有显著性差异(P<0.05);而用麻黄碱加甘草酸配制成的溶液无退热效果。作者认为应从生物—配位无机化学的多金属—多配体体系的观点去研究某些传统中医药方剂。

Ontogeny and Metamorphic Patterns of Female Reproductive Organs of Ephedra sinica Stapf (Ephedraceae)

Ontogeny and metamorphic. development of female reproductive organs in Ephedra sinica Stapf were surveyed. At the end of April, female cones began to initialize from the vegetative buds. Pollination occurred in mid-May and seeds matured at the beginning of July. The ontogenetic pattern of female reproductive organs of E. sin ca is basically similar to that of E. distachya L. The foliar nature of the outer envelope of the ovule in Ephedra is corroborated. Reduction of ovule number as a tendency of speciation in the genus is substantiated by the occurrence of tri-ovulate cones coupled with comprehensive characters of the genus. The metamorphic patterns as well as the leaf nature of the outer envelope indicate that female cones of Ephedra are compound while the female reproductive units of the ovulate cone have been reduced from secondary reproductive shoots similar to those of cordaites by phylogenetic transformation, fusion and reduction. Each fertile bract together with its axillary female reproductive unit composed the Bract Scale and Seed Scale Complex of Ephedra.

麻黄、桂枝及其药对中矿质元素初级形态含量的测定

采用火焰原子吸收分光光度法测定了麻黄(Herbal ephedra)、桂枝(Ramulus cinnamomi)及其药对中Ca、Cr、Fe、Mg、Mn、Zn等6种矿质元素的含量。结果表明,麻黄、桂枝的Ca含量较高,其次是Mg和Fe;Mg的提取率最高,分别为64.74%和45.88%,药对的提取率均介于两种单味药材之间。对这两种药材及其药对中矿质元素初级形态的研究,为进一步探讨矿质元素与药效之间的关系提供参考。

麻黄内生真菌的初步研究

从宁夏回族自治区贺兰山高盐碱地区采集的麻黄样本中分离得到7株内生真菌,体外抗氧化活性测定表明,所分离到的内生真菌中有5株具有较高的抑制羟基自由基能力;有5株菌的发酵液具有中等强度的总抗氧化能力。在抗菌活性测定中,所分离到的7株麻黄内生真菌对细菌指示菌大肠杆菌和金黄色葡萄球菌均表现出一定的抑菌作用,但对真菌指示菌炭疽杆菌和黑曲霉均未检测到抑菌活性。

中麻黄新疆产地生态气候适宜性分析

以实地调查采样点、标本查阅和文献研究为基础,结合《中药材产地适宜性分析地理信息系统-Ⅰ》进行综合分析中麻黄在新疆的适宜生长区域,为中麻黄规范化种植、科学选址及合理规划生产布局提供依据。结果表明,中麻黄全疆广为分布,适宜区为吐鲁番盆地一带及准噶尔盆地北部的库车、轮台、柯坪、阿图什、阿合奇及阿克陶县。

A fossil-calibrated relaxed clock for Ephedra indicates an Oligocene age for the divergence of Asian and New World clades and Miocene dispersal into South America

Ephedra comprises approximately 50 species, which are roughly equally distributed between the Old and New World deserts, but not in the intervening regions （amphitropical range）. Great heterogeneity in the substitution rates of Gnetales （Ephedra, Gnetum, and Welwitschia） has made it difficult to infer the ages of the major divergence events in Ephedra, such as the timing of the Beringian disjunction in the genus and the entry into South America. Here, we use data from as many Gnetales species and genes as available from GenBank and from a recent study to investigate the timing of the major divergence events. Because of the tradeoff between the amount of missing data and taxon/gene sampling, we reduced the initial matrix of 265 accessions and 12 loci to 95 accessions and 10 loci, and further to 42 species （and 7736 aligned nucleotides） to achieve stationary distributions in the Bayesian molecular clock runs. Results from a relaxed clock with an uncorrelated rates model and fossil-based calibration reveal that New World species are monophyletic and diverged from their mostly Asian sister clade some 30 mya, fitting with many other Beringian disjunctions. The split between the single North American and the single South American clade occurred approximately 25 mya, well before the closure of the Panamanian Isthmus. Overall, the biogeographic history of Ephedra appears dominated by long-distance dispersal, but finer-scale studies are needed to test this hypothesis.

Identification of Two Cold Water-Soluble Polysaccharides from the Stems of Ephedra sinica Stapf

Two polysaccharides (ESP-A1 and ESP-A2) were isolated from the cold water extract of Ephedra sinica Stapf and purified through ethanol precipitation, deproteinization and by ion exchange and gel-filtration chromatography. Their molecular weight was determined using high performance size exclusion chromatog-raphy and evaporative light scattering detector (HPSEC-ELSD) and their monosaccharide composition was analyzed by high performance capillary electrophoresis (HPCE) based on pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP). It was shown that ESP-A1 consisted of xylose, arabinose, glucose, mannose and galactose and ESP-A2 consisted of xylose, arabinose, rhamnose and galactose, in a molar ratio (%) of 3.2: 61.1: 11.1: 12.9: 11.6 and 20.6: 67.7: 5.0: 6.7, respectively. The molecular weights (Mw) of ESP-A1 and ESP-A2 were 5.83 × 104 Da and more than 200 × 104 Da, respectively. To the best of our knowledge, two neutral polysaccharides are now being reported for the first time in this study.

Effects of ephedra asarum aconite decoction on Th1/Th2 balance in two animal models of allergic rhinitis

Objective:Ephedra asarum aconite decoction is used in the clinical treatment of allergic rhinitis.We investigated the effects of ephedra asarum aconite decoction on Th1/Th2 balance in two animal models of allergic rhinitis and in activated T cells in vitro.Methods:Animal models of simple and compound kidney yang deficiency types of allergic rhinitis were treated with ephedra asarum aconite decoction to observe the efficacy of this traditional Chinese medicine formula on allergic symptoms,serum cytokines,and tissue RNA levels.We also cultured and activated T cells from the mouse.After treatment with ephedra asarum aconite decoction,we tested the proliferation and differentiation of T cells.Results:Ephedra asarum aconite decoction relieved allergy symptoms in both types of animal models,and was more effective in the compound kidney yang deficiency model In addition to its anti-allergic efficacy,ephedra asarum aconite decoction significantly improved the health status of animals in the compound kidney yang deficiency groups,significantly made their bodyweight rising up,and significantly increased the T-bet mRNA/GATA-3 mRNA ratio.Moreover,ephedra asarum aconite decoction promoted the proliferation of activated T cells and inhibited the differentiation to Th2 cells in vitro.Conclusions:Ephedra asarum aconite decoction regulates the balance of Th1/Th2 to ameliorate allergic conditions.

Effects of the ephedra alkaloid methylephedrine on the basal evoked potential transportation and the long-term potentiation (LTP) in the rat hippocampal dentate granule cells in vivo

Objective The effect of the ephedra/ephedrine alkaloid methylephedrine(dl-methylephedrine hydrochloride for testing in this paper)on cognitive related synaptic plasticity was investigated by recording extracellular field evoked potentials and its LTP in hippocampal dentate granule cells in urethane-anaesthetized rats in vivo.Methods Single pathway recording of evoked field potentials was made from the dentate granule cells of hippocampal hemisphere in response to stimulation of the ipsilateral medial perforant path(MPP).Two parameters,the amplitude of population spike(PS amplitude)and the latency of the PS,were employed to evaluate the effects of drug on the overall changes in cellular responses.Results The present study show that methylephedrine 90 mg·kg-1 intraperitoneally,about 1/3 LD50,could increase the latency of the PS in hippocampal dentate granule cells by constant single stimulation of the MPP as the basal ransportation.However,the 30 mg·kg-1 and 10 mg·kg-1 dosage had no effect on the latency,and there are no influences of PS amplitude for all examinational groups.The methylephedrine 90 mg·kg-1 group significant enhanced the development of amplitude LTP in hippocampal dentate granule cells that induced by 60 Hz,60 pulses conditional tetanus in medial perforant path area.Also,the 30 mg·kg-1 group can promoted the maintenance of LTP induced by this tetanus,but no promotion on PS amplitude LTP appeared in this dosage and no any changes been found in 10 mg·kg-1 dosage group.Conclusions The ephedra/ephedrine alkaloid methylephedrine can modulate the synaptic plasticity in the lateral perforant path.A possible mechanism of methylephedrine on hippocampal LTP is been discussed.

Herbivory Effects on <i>Ephedra</i>spp. in the Chihuahuan Desert

Two species of Ephedra: E. trifurca and E. torreyana inhabit shrub and grassland habitats in the northern Chihuahuan Desert. E. torreyana is limited to black grama grasslands where grasses are taller than the shrub. E. torreyana is heavily browsed by vertebrates and E. trifurca is browsed during some years. We established an experiment with cylindrical exclosures that excluded rabbits and rodents, rabbits but accessible to rodents, for comparison with E. torreyana plants available to all herbivores. Plants accessible to all vertebrate herbivores were significantly smaller with shorter stem lengths than plants in exclosures. We concluded that E. torreyana in black grama grassland are largely hidden from vertebrate herbivores and that intense herbivory reflects the degraded state of the study site which makes the E. torreyana evergreen shrubs apparent to vertebrates.

野生植物资源——草麻黄

草麻黄(Ephedra sinica)属麻黄科草本状小灌木,别名麻黄、麻黄草,蒙古名哲格日根讷。草麻黄株高20—40cm,木质茎短或匍匐状,小枝直立或弯曲,叶退化成膜质鞘状,2裂。小枝绿色,代替叶进行光合作用、呼吸作用和蒸腾作用。雄球花为复穗状,密集于节上;雌球花单生当年枝条顶端或老枝腋中。5—6月开花,8—9月雌球花成熟,

Maintenance of Ephedra alata Seeds Viability via Storage Containers

Sequential incubation of seed samples yielded 20 fungal species belonging to 13 genera. The prevalent genera were Aspergillus (A. flavus and A. parasiticus), Fusarium (F. moniliforme and F. oxysporum) and Penicillium (Penicillium sp.). Such seedborne fungi differ in their colonization in different parts of seeds with most of them are colonized in seed coat, endosperm, and embryo of the seed. The usage of different storage containers for storing seeds indicated that the cotton cloth bags were the most favorable ones as they maintain seed moisture content (SMC) below the critical level resulting in minimum seed deterioration compared with other seed storage containers.

Action mechanism of Ephedra in treating allergic rhinitis based on network pharmacology

Objective:To study the mechanism of ephedra in treating allergic rhinitis by network pharmacology.Methods:the chemical components and corresponding targets of ephedra were searched and screened by TCMSP database.With"allergic rhinitis"as the key word,disease-related genes were searched in GeneCards and OMIM database.Using Venny2.1.0 mapping,Venn diagram and drug-disease common targets,network visualization software Cytoscape 3.6.1 track production elements-targets-disease network diagram,String database build their proteins interact with each other(PPI),and using R language to do gene ontology(GO)functional enrichment and pathway analysis and kyoto encyclopedia of genes and genome(KEGG)pathway enrichment analysis.Results:ephedrine was screened for 136 efficacy components and 88 common targets,and was predicted to intervene and regulate allergic rhinitis through AGE-RAGE signaling pathway in diabetic complications,Fluid shear stress and atherosclerosis,IL-17 signaling pathway,TNF signaling pathway and other signaling pathways in diabetic complications.Conclusion:preliminary prediction of the target and pathway of ephedra in the treatment of allergic rhinitis through network pharmacology can provide direction and evidence for the next experiment and clinic.

Study on the mechanism of action of Ephedra Herba Decoction against influenza A virus based on network pharmacology

Objective To predict the main bioactive components and potential mechanisms of Ephedra Herba Decoction(Chinese Pinyin abbreviated as MHT)in treating influenza A virus(IAV)based on network pharmacology.Methods Multiple online databases were used to search and screen out the active components from MHT,the related targets of active components of MHT and the genes related to IAV.Search the corresponding genes name of target through UniProt database.Cytoscape 3.7.2 was used to construct the drug-component-target network diagram.Venn diagram was used to screen the intersection genes of the active components corresponding to the target and disease-related genes,and the intersection genes were imported into the STRING Database Online platform to obtain the protein-protein interaction(PPI)network.Then,the PPI network was imported into Cytoscape 3.7.2 to obtain the core targets.Finally,Wei Sheng Xing(http://www.bioinformatics.com.cn/)was used to do GO function enrichment analysis and KEGG signaling pathway enrichment analysis for the intersection genes,and the results of GO and KEGG were visualized.Results A total of 116 active components and 253 potential targets were screened from MHT.Quercetin,Kaempferol,and Luteolin are the main active components,and AKT1,TNF,TP53,IL6,and JUN are the core targets of 253 potential targets.There are 2906 targets of influenza A,including 121 intersection genes.Enrichment analysis showed that there were 131 entries of molecular function,89 entries of cell component,1645 entries of biological process and 195 entries of signaling pathway.PI3K-Akt,MAPK,and JAK-STAT were the main signaling pathways.Conclusion MHT plays an important role in the prevention and treatment of influenza A by acting on multiple targets and multiple signaling pathways.

Extraction process in Chuantieling(CTL) based on the quality by design(QbD) concept

Objective This study aimed to design and optimize the water extraction process for Chuantieling(喘贴灵,CTL)based on the concept of quality by design(QbD).Methods The single-factor experiments were used to select the best experimental points of CTL water extraction.On this basis,the transfer rate of ephedrine hydrochloride and sin-apine thiocyanate,and the yield of the extract were evaluated as the evaluation indicators.The liquid-solid ratio,extraction time,and pH value were selected as the main factors to carry out the Box-Behnken design(BBD).A mathematical model of the critical process parameters(CPPs)and critical quality attributes(CQAs)was established,the interaction between CQAs and CPPs was examined,and the CTL extraction process design space was established and optimized,which guaranteed the stability of the process.The high performance liquid chro-matography(HPLC)was used to analyze the main active compounds in the CTL extract.Results Through single-factor experiments,the best experimental parameters were found to be a liquid-solid ratio of 8∶1,extraction time of 90 min,pH value of 4,and extraction times of three.The experimental variance analysis results of the BBD showed that the P value of the re-gression model was less than 0.05,and the lack-of-fit value was greater than 0.01,indicating that the constructed model had good predictive ability.The operating space of the CPPs of the CTL water extraction process was combined with the actual production situation.In the pro-duction situation,the best extraction process was nine times of water addition,72 min of ex-traction time,and 4.5 of pH value.The HPLC results showed that the peak areas of ephedrine hydrochloride,sinapine thiocyanate,tetrahydropalmatine,methyl eugenol,cinnamaldehyde,and 6-gingerol in the CTL extract accounted for 0.94%,14.32%,0.78%,31.23%,0.34%,and 0.44%of the total peak area,respectively.Conclusion The water extraction process design space of CTL based on QbD was conducive to actual production operations,ensuring the stability of the process.