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RECENT SITUATION AND PROSPECTS OF THE STADY OF EUCOMMIA ULMOIDES
杜仲研究的现状与展望

杨峻山,姜声虎
自然资源学报 , 1997,
Abstract: This paper generally describes the present study state of Eucommia ulmoides in the aspects of resources, cultivation, chemical constituents, medicinal activity and clinical application. Furthermore, the prospects of its exploitation and utilization are forecastad.
Novel phytoandrogens and lipidic augmenters from Eucommia ulmoides
Victor YC Ong, Benny KH Tan
BMC Complementary and Alternative Medicine , 2007, DOI: 10.1186/1472-6882-7-3
Abstract: The extracts of E. ulmoides were tested using in-vitro reporter gene bioassays and in-vivo animal studies. Key compounds responsible for the steroidogenic effects were isolated and identified using solid phase extraction (SPE), high performance liquid chromatography (HPLC), thin layer chromatography (TLC), gas chromatography-mass spectroscopy (GC-MS), electron spray ionisation-mass spectroscopy (ESI-MS) and nuclear magnetic resonance (NMR).The following bioactivities of E. ulmoides were found: (1) a phenomenal tripartite synergism exists between the sex steroid receptors (androgen and estrogen receptors), their cognate steroidal ligands and lipidic augmenters isolated from E. ulmoides, (2) phytoandrogenic activity of E. ulmoides was mediated by plant triterpenoids binding cognately to the androgen receptor (AR) ligand binding domain.In addition to well-known phytoestrogens, the existence of phytoandrogens is reported in this study. Furthermore, a form of tripartite synergism between sex steroid receptors, sex hormones and plant-derived lipids is described for the first time. This could have contrasting clinical applications for hypogonadal- and hyperlipidaemic-related disorders.The androgen receptor (AR) plays a pivotal role in human (both male and female) physiology such as skeletal muscle development, bone density, fertility and sex drive [1,2]. The α and β estrogen receptors (ERs), likewise, have fundamental impact on the sex hormone-mediated physiological milieu. Conversely, over-active sex steroid (androgen and estrogen) receptors have been linked to increased risks of hormone-sensitive tumours such as prostate and breast cancers. Availability and binding of cognate ligands to the ligand binding domain (LBD) of the sex steroid receptors are required for the proportionate expression of specific genes responsible for such sex hormone-mediated processes [3,4].Vegetative foods such as the legumes, particularly soybean (Glycine max), contain phytoestrogens that modu
Extraction and determination of major hypotensive compounds in bark of Eucommia ulmoides Oliv.
Xiao Jianbo,Wei Xinlin,Wang Yuanfeng
Archives of Biological Sciences , 2009, DOI: 10.2298/abs0904811x
Abstract: A reversed-phase liquid chromatographic method was developed for the quantitative determination of three major hypotensive compounds, namely geniposidic acid, chlorogenic acid, and geniposide in the bark of Eucommia ulmoides. Soxhlet extraction of GPA, GPS, and CA from E. ulmoides was optimized according to the Taguchi experimental design. Maximum global yields were obtained using the following conditions: extraction temperature, 80°C; extraction time, 1 h; number of extractions, three; solvent volume, 16 ml/g of sample; and 50% ethanol concentration in water. Optimal conditions of separation and detection were achieved on a Diamonsil ODS C18 column (150 mm × 4.6 mm, 5 μm) with a linear gradient of methanol and 0.04% aqueous phosphoric acid (v/v) at a flow rate of 1.0 ml/min and detection wavelength of 240 nm. All calibration curves showed good linearity (r2 > 0.999) within test ranges. The relative deviation of this method was less than 3% for intra- and inter-day assays, and the recovery percentage of the method was 95-104%, with a relative standard deviation (R.S.D.) of less than 5%. The current assay method was used for quantitative determination of geniposidic acid, chlorogenic acid, and geniposide in five samples of E. ulmoides with different age. The results indicate that the developed method could be readily utilized as a quality control method in working with E. ulmoides.
Comparison of extraction methods for the hypotensive drugs from Eucommia ulmoides
Cao Hui
Archives of Biological Sciences , 2010, DOI: 10.2298/abs1003725c
Abstract: Extraction methods using Soxhlet extraction (SE), enzyme-assisted aqueous extraction (EE), semi-bionic extraction (SBE) and supercritical fluid extraction (SFE) were evaluated for the yields of geniposidic acid (GPA) and geniposide (GPS) from the bark of Eucommia ulmoides. The results showed that the yields of GPA and GPS attained by EE and SBE were highest, respectively. Compared with various extraction methods for the extraction of GPA and GPS, the SBE was more efficient than other methods. There was no organic solvent used in SBE. The pH values of semi-bionic extraction were the same as in the human body. The results indicate that SBE can be used for large-scale and efficient extraction of GPA and GPS from plant materials. The time taken by SFE was longer than other methods, but the yields of GPA and GPS were the lowest. .
Identification of a Male-Specific Amplified Fragment Length Polymorphism (AFLP) and a Sequence Characterized Amplified Region (SCAR) Marker in Eucommia ulmoides Oliv.  [PDF]
Da-Wei Wang,Yu Li,Zhou-Qi Li
International Journal of Molecular Sciences , 2011, DOI: 10.3390/ijms12010857
Abstract: Eucommia ulmoides Oliv. is a dioecious species. Currently, there is no method to identify the sex during the juvenile stage that lasts a relatively long time. This study aimed to develop a sex specific Amplified Fragment Length Polymorphism (AFLP) marker for Eucommia ulmoides Oliv. Of a total of 64 AFLP primer combinations screened, primer combination E-ACA/M-CTT produced a 350 bp male-specific marker. This 350 bp AFLP marker was converted into a 247 bp Sequence Characterized Amplified Region (SCAR) marker. Results suggest that the SCAR marker can be utilized for early sexual identification in Eucommia ulmoides Oliv., and it will greatly facilitate future breeding?programs.
Eucommia ulmoides Oliver Extract, Aucubin, and Geniposide Enhance Lysosomal Activity to Regulate ER Stress and Hepatic Lipid Accumulation  [PDF]
Hwa-Young Lee, Geum-Hwa Lee, Mi-Rin Lee, Hye-Kyung Kim, Nan-young Kim, Seung-Hyun Kim, Yong-Chul Lee, Hyung-Ryong Kim, Han-Jung Chae
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0081349
Abstract: Eucommia ulmoides Oliver is a natural product widely used as a dietary supplement and medicinal plant. Here, we examined the potential regulatory effects of Eucommia ulmoides Oliver extracts (EUE) on hepatic dyslipidemia and its related mechanisms by in vitro and in vivo studies. EUE and its two active constituents, aucubin and geniposide, inhibited palmitate-induced endoplasmic reticulum (ER) stress, reducing hepatic lipid accumulation through secretion of apolipoprotein B and associated triglycerides and cholesterol in human HepG2 hepatocytes. To determine how EUE diminishes the ER stress response, lysosomal and proteasomal protein degradation activities were analyzed. Although proteasomal activity was not affected, lysosomal enzyme activities including V-ATPase were significantly increased by EUE as well as aucubin and geniposide in HepG2 cells. Treatment with the V-ATPase inhibitor, bafilomycin, reversed the inhibition of ER stress, secretion of apolipoprotein B, and hepatic lipid accumulation induced by EUE or its component, aucubin or geniposide. In addition, EUE was determined to regulate hepatic dyslipidemia by enhancing lysosomal activity and to regulate ER stress in rats fed a high-fat diet. Together, these results suggest that EUE and its active components enhance lysosomal activity, resulting in decreased ER stress and hepatic dyslipidemia.
Overexpression of an isopentenyl diphosphate isomerase gene to enhance trans-polyisoprene production in Eucommia ulmoides Oliver  [cached]
Chen Ren,Harada Yoko,Bamba Takeshi,Nakazawa Yoshihisa
BMC Biotechnology , 2012, DOI: 10.1186/1472-6750-12-78
Abstract: Background Natural rubber produced by plants, known as polyisoprene, is the most widely used isoprenoid polymer. Plant polyisoprenes can be classified into two types; cis-polyisoprene and trans-polyisoprene, depending on the type of polymerization of the isoprene unit. More than 2000 species of higher plants produce latex consisting of cis-polyisoprene. Hevea brasiliensis (rubber tree) produces cis-polyisoprene, and is the key source of commercial rubber. In contrast, relatively few plant species produce trans-polyisoprene. Currently, trans-polyisoprene is mainly produced synthetically, and no plant species is used for its commercial production. Results To develop a plant-based system suitable for large-scale production of trans-polyisoprene, we selected a trans-polyisoprene-producing plant, Eucommia ulmoides Oliver, as the target for genetic transformation. A full-length cDNA (designated as EuIPI, Accession No. AB041629) encoding isopentenyl diphosphate isomerase (IPI) was isolated from E. ulmoides. EuIPI consisted of 1028 bp with a 675-bp open reading frame encoding a protein with 224 amino acid residues. EuIPI shared high identity with other plant IPIs, and the recombinant protein expressed in Escherichia coli showed IPI enzymatic activity in vitro. EuIPI was introduced into E. ulmoides via Agrobacterium-mediated transformation. Transgenic lines of E. ulmoides overexpressing EuIPI showed increased EuIPI expression (up to 19-fold that of the wild-type) and a 3- to 4-fold increase in the total content of trans-polyisoprenes, compared with the wild-type (non-transgenic root line) control. Conclusions Increasing the expression level of EuIPI by overexpression increased accumulation of trans-polyisoprenes in transgenic E. ulmoides. IPI catalyzes the conversion of isopentenyl diphosphate to its highly electrophilic isomer, dimethylallyl diphosphate, which is the first step in the biosynthesis of all isoprenoids, including polyisoprene. Our results demonstrated that regulation of IPI expression is a key target for efficient production of trans-polyisoprene in E. ulmoides.
基于RNA-Seq的杜仲转录组微卫星特征分析
Microsatellites characteristics of transcriptomic sequences from Eucommia ulmoides Oliv.based on RNA-Seq
 [PDF]

冯延芝,李芳东,魏琦琦,莫文娟,王璐,黄地歌,傅建敏
- , 2016,
Abstract: 对杜仲(Eucommia ulmoides)国审良种‘华仲6号’和‘华仲10号’花后70和160 d的种仁共4个样本进行转录组测序,对测序数据进行组装和功能注释分类,并对转录组获得的单基因簇(unigene)进行微卫星特征分析。利用新一代高通量测序技术Illumina HiSeqTM 2000对杜仲样品进行转录组测序,采用软件Trinity进行组装;利用BLAST软件将unigene序列分别与Nr、GO、COG和KEGG等数据库比对分析;利用MISA软件对转录组的96 469条unigenes进行SSR搜索。结果表明:转录组测序分析,共得到72 791 399个高质量的序列读取片段(Clean reads),包含了14 702 548 161个的碱基序列(bp)信息。对reads进行序列组装,共获得96 469个平均长度为690 bp的unigene,序列信息量达到了66.56 Mb。同源性分析结果显示,有49 856个与其它物种同源的unigenes得到注释,占All-unigene的51.68%。将杜仲转录组中的unigene与GO数据库进行比对分析,根据其功能可将注释到的38 983条unigene分成3大类(细胞组分、分子功能和生物学过程)56个分支;根据COG功能可将注释的14 796条unigene基因划分成25个类别;KEGG数据库作为参照,可将注释到的11 260条unigene定位到117个代谢途径分支;SSR位点搜索结果显示,96 469条unigenes中共包含9 621个完整型SSR位点,占总SSR位点的84.14%。完整型SSR位点共包含55种重复基元,其中出现频率最高的重复基序类型为单核苷酸重复中的A/T(4 597个),其次是AG/CT(2 597个)、AT/AT(439个)。
The transcriptomes of Eucommia ulmoides Oliv. kernels of 70 and 160 d after flowering in varieties ‘Huazhong 6 and 10’ were sequenced.The transcriptome data was assembled and classified by function,and microsatellites characteristics from obtained unigenes and analyzed.The Illumina HiSeqTM 2000,a new generation of high-throughput sequencing technology was used to sequence the transcriptomes of kernels of assembled by software Trinity.The unigenes were annotated according to Nr,GO,COG and KEGG category by BLAST searches.A total of 72 791 399 clean reads fragment including 14 702 548 161 bp in sequence information were generated,and then de novo assembly generated a total of 96 469 unigenes with an average length of 690 bp,which contains 66.56 Mb in sequence information.Among them,49 856 unigenes accounted for 51.68% were annotated by BLAST searches.All 38 983 annotated unigenes according to GO were divided into three categories (cellular components,molecular function and biological processes) of 56 branches by gene ontology;14 796 annotated unigenes based on COG were grouped into 25 functional categories;KEGG pathway analysis presented that 11 260 annotated unigenes were divided into 117 classes according to its function.There were 9 621 complete SSR located in 96 469 unigenes,which accounted for 84.14% of the total SSR.The complete SSR included 55 frequent motifs,and the highest repeat of complete SSR type was A/T (4 597),following by AG/CT (2 597)、AT/AT (439).The characteristics of SSRs can provide useful information for the analysis of genetic polymorphism and map structure in E.ulmoides.
Seasonal Difference in Antioxidant Capacity and Active Compounds Contents of Eucommia ulmoides Oliver Leaf  [PDF]
Qiang Zhang,Yinquan Su,Jingfang Zhang
Molecules , 2013, DOI: 10.3390/molecules18021857
Abstract: Leaf of Eucommia ulmoides Oliver (EU) is a Traditional Chinese Medicine and a functional food in China. Antioxidant contents of EU leaves, which were collected monthly during the period of May–October in three years, were determined. Samples’ antioxidant capacity was characterized by DPPH radical scavenging activity, hydroxyl radical scavenging activity, ferrous chelating ability, and antioxidant capacity in linoleic acid emulsion and in rapeseed oil assays. The results showed that contents of some active compounds and antioxidant activity were related to a certain time of the year. Samples collected in August showed high content of phenolics, and the samples collected in May contained higher amount of flavonoids than other samples. Leaves collected in May or June exhibited high contents of rutin, quercetin, geniposidic acid and aucubin. The August leaves showed stable and high DPPH radical scavenging activity, and ferrous chelating ability. May samples showed strong inhibitory effects on oxidation of rapeseed oil and linoleic acid. The DPPH radical scavenging activity was related to the total phenolics content. Flavonoids played an important role in the inhibitory effects on rapeseed oil and linoleic acid oxidation. Therefore, August and May were indicated as the best months to harvest EU leaves for industry.
Eucommia ulmoides Cortex, Geniposide and Aucubin Regulate Lipotoxicity through the Inhibition of Lysosomal BAX  [PDF]
Geum-Hwa Lee, Mi-Rin Lee, Hwa-Young Lee, Seung Hyun Kim, Hye-Kyung Kim, Hyung-Ryong Kim, Han-Jung Chae
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0088017
Abstract: In this study we examined the inhibition of hepatic dyslipidemia by Eucommia ulmoides extract (EUE). Using a screening assay for BAX inhibition we determined that EUE regulates BAX-induced cell death. Among various cell death stimuli tested EUE regulated palmitate-induced cell death, which involves lysosomal BAX translocation. EUE rescued palmitate-induced inhibition of lysosomal V-ATPase, α-galactosidase, α-mannosidase, and acid phosphatase, and this effect was reversed by bafilomycin, a lysosomal V-ATPase inhibitor. The active components of EUE, aucubin and geniposide, showed similar inhibition of palmitate-induced cell death to that of EUE through enhancement of lysosome activity. Consistent with these in vitro findings, EUE inhibited the dyslipidemic condition in a high-fat diet animal model by regulating the lysosomal localization of BAX. This study demonstrates that EUE regulates lipotoxicity through a novel mechanism of enhanced lysosomal activity leading to the regulation of lysosomal BAX activation and cell death. Our findings further indicate that geniposide and aucubin, active components of EUE, may be therapeutic candidates for non-alcoholic fatty liver disease.
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