Comparative analysis of chemical components between barks and leaves of Eucommia ulmoides Oliver

The chemical components of the essential oils in the barks and leaves of Eucommia ulmoides Oliver were analyzed and compared by chromatograms and mass spectra technique, heuristic evolving latent projections (HELP), alternative moving window factor analysis (AMWFA) algorithms and normalization method based on the peak areas; the flavones in the barks and leaves of Eucommia ulmoides Oliver were separated on an ODS column by gradient elution carried out with the flow phase consisting of water, methanol and phosphoric acid (0.1%), and their contents were quantitatively determined by standard curve method and diode array detection (DAD) at 362 nm. The results show that 68 and 73 compounds respectively from essential oils of the barks and leaves of Eucommia ulmoides Oliver are identified, and there are 33 mutual compounds among 108 compounds determined. The total contents of these volatile components of the two samples possess 92.9% and 97.75% of the gross of the relevant essential oils, respectively; the contents of the rutin, quercetin and kaempferol in the barks and leaves of Eucommia ulmoides Oliver are 0.016 9, 0.003 6, 0.002 1 and 0.064 4, 0.030 2, 0.010 0 mg/g, respectively, and the determination recoveries are 95.2%-106.2%. The comparative analysis shows that for the barks and leaves of Eucommia ulmoides Oliver, there are significant differences in their components of the relevant essential oils and flavones.

Chemical constituents,biological functions and pharmacological effects for comprehensive utilization of Eucommia ulmoides Oliver

Eucommia ulmoides Oliver is a native plant and valuable tonic Chinese medicine in China with a long history,great economic value and comprehensive development potential.Traditionally,the comprehensive utilization rate of E.ulmoides Oliv.is still very low,only bark has been used as medicine and other parts of Eucommia ulmoides Oliv.cannot be fully utilized,even the leaves have been well utilized in food products in Japan in the past decades.In order to improve the comprehensive utilization efficiency of E.ulmoides Oliv.,in this review,we summarized the varieties and contents of main active compounds,biological functions and pharmacological effects in different parts of E.ulmoides Oliv.The findings suggest that other parts of E.ulmoides Oliv.could replace the bark of E.ulmoides Oliv.to some extent besides of their respective applications.The unique and extensive physiological functions between different parts of E.ulmoides Oliv.indicate that the comprehensive utilization of E.ulmoides Oliv.has a wide space to develop,which is also an effective way to protect E.ulmoides Oliv.resources and improve its the utilization rate.

The Extraction of Eucommia Ulmoides Oliv Polysaccharides and Its Protective Effect on Liver of Clophasphamidecy Injured Mice

Objective:To study the influenceof eucommia polysaccharide on the mice' liver damaged by clophasphamidecy (CY).Methods:Injecting CY build mice liver damage model,eucommia polysaccharide given different doses, measured blood serum ALT,AST and the liver's SOD,MDA. Results:After the injection CY,blood serum ALT,AST and the MDA of liver rise and the SOD of liver reduce comparedwith the blank group. The eucommia polysaccharide can improve these index.Conclusion:The Eucommia polysaccharide may protect the mice' liver damaged by CY.

Isolation of a novel antifungal peptide from the bark of Eucommia ulmoides Oliv

A novel Eucommia antifungal peptide, named EAFP3, was isolated from the bark of Eucommia ul- moides by NaC1 extract, gel filtration and reverse phase high performance liquid chromatography. The molecular mass of EAFP3 is 4157.3 Da, and its partial amino acid sequence is -. LYQQLIAGITLNK.-. EAFP3 exerts an inhibitory activity against Candida albicans in vitro and the drug concentration required for 50% growth inhibition （IC50） is 31.25μg/mL.

Cloning and Characterization of EuGID1 in Eucommia ulmoides Oliver

Gibberellic acid controlled the key developmental processes of the life cycle of landing plants,and regulated the growth and development of plants.In this study,a novel gibberellin receptor gene EuGID1 was obtained from Eucommia ulmoides Oliver.The cDNA of EuGID1 was 1556 bp,and the open reading frame was 1029 bp,which encoded 343 amino acids.EuGID1 had the homology sequence with the hormone-sensitive lipase family.Amino acid sequence alignment confirmed EuGID1 protein had the highest homology with the GID1 protein of Manihot esculenta.EuGID1 was located in the nucleus and cell membrane and had expression in four plant organs.Overexpression of EuGID1 in transgenic Arabidopsis plants promoted plant elongation and increased siliques yield.

Extraction Process of Total Flavonoids from Eucommiae Cortex and Its in vitro Antioxidant Activity

[Objectives]To optimize the extraction process of total flavonoids from Eucommiae Cortex,establish the extraction and content determination methods of its medicinal materials,and study its in vitro antioxidant activity.[Methods]The total flavonoids from Eucommiae Cortex were extracted by reflux extraction method,and the effects of extraction method,extraction solvent concentration,extraction volume,and extraction time on the total flavonoids content of the medicinal materials were explored through the single factor experiment.Orthogonal experiment was carried out to optimize the extraction process conditions,and the optimal extraction process for total flavonoids from Eucommiae Cortex was screened out.The total antioxidant capacity index was used to determine the free radical scavenging ability of the total flavonoids from Eucommiae Cortex.[Results]The optimal extraction process of total flavonoids from Eucommiae Cortex was 50%ethanol,1∶40 solid-to-liquid-ratio,and 1.5 h reflux extraction time.Through the antioxidant experiment in vitro,it is found that the total flavonoids from Eucommiae Cortex have a higher scavenging ability for the total antioxidant capacity,and there is a certain positive correlation with the mass concentration of total flavonoids.[Conclusions]This method can effectively determine the total flavonoids content of Eucommiae Cortex medicinal material,and provide a certain scientific basis for the quality standard research of the medicinal material.The total flavonoids from Eucommiae Cortex have good in vitro antioxidant activity.And the method for extracting total flavonoids from Eucommiae Cortex medicinal material in this experiment has good repeatability,high stability and feasibility.

杜仲协同拟茎点霉XP-8合成木脂素类的条件优化及其功能活性

对杜仲协同拟茎点霉XP-8生物合成松脂醇(Pinoresinol,Pin)、松脂醇单葡萄糖苷(Pinoresinol-4-O-β-Dglucopyranoside,PMG)和松脂醇二葡萄糖苷(Pinoresinol diglucoside,PDG)的条件进行优化,研究培养方式、杜仲添加量和静息处理时间对Pin、PMG和PDG产量的影响,并对相关产物的抗氧化性和抑癌性进行研究。结果表明,3%杜仲PDB培养基培养菌体并静息处理5 d时Pin、PMG和PDG产量较高。该条件下代谢产物液对DPPH和ABTS+自由基的清除能力显著高于杜仲空白培养基,说明发酵过程中产生了增强抗氧化功能的物质。其还能抑制肝癌细胞HepG-2的增殖,通过纯品验证活性说明,Pin是其中主要抑制癌细胞的活性成分,代谢产物液抑制肝癌细胞的作用是一种协同效应。

杜仲翅果果实形态和含胶量与产地环境相关性分析

该研究选择我国7个杜仲产地的杜仲为研究对象,采用随机取样的方法,利用SPSS软件进行统计分析。结果表明:不同产地果实形态和含胶量存在显著差异,贵州剑河最高为13.16%,河北保定最低为9.34%,含胶量与所在环境气候条件密切相关,纬度越高,年日照时数越长,翅果含胶量越低;无霜期越长,翅果含胶率越高。

杜仲对黄曲霉的抑制及在养殖中的有益作用

饲料中黄曲霉污染严重制约了饲料业和养殖业的高质量发展,在我国目前饲料端全面禁抗背景下,天然植物原料作为理想的抗生素替代品被推崇。杜仲是我国传统的药食两用植物资源,将其提取物或原粉添加到饲料中可有效遏制黄曲霉污染,并提高畜禽品质,是理想的饲料替抗物。文章阐述了杜仲提取物及其活性成分对黄曲霉的抑制作用以及在养殖中的有益作用,旨在为其在饲料行业中的应用推广提供参考。

Eucommia ulmoides Oliv.antagonizes H_2O_2-induced rat osteoblastic MC3T3-E1 apoptosis by inhibiting expressions of caspases 3,6,7,and 9

Eucommia ulmoides Oliv.(EuO),also known as Duzhong,native to China,has been reported to have antioxidative function,but its cellular mechanism is not fully examined yet.We investigated inhibitory effects of EuO leaf ethanol extracts on H2O2-induced apoptosis in rat osteoblastic MC3T3-E1 cells and underlying mechanisms.Locally-grown Duzhong leaves were extracted with ethanol.MC3T3-E1 cells were treated with EuO (6.25,12.5,25,50,and 100 μg/ml) for 24 h,and then H2O2 (800 μmol/L) for an additional 24 h.Cell survival rate,percentage of apoptosis,and expressions of caspases 3,6,7,and 9 were examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,microscopic analysis,Western blotting,and reverse transcription polymerase chain reaction (RT-PCR).The final EuO leaf ethanol extract powder was detected to contain caffeotannic acid at 58 mg/g and geniposide at 3.45 mg/g by high performance liquid chromatography (HPLC).EuO remarkably restrained cell oxidative damage and increased cell survival rate in a dose-dependent manner: 0 μg/ml,0.21;6.25 μg/ml,0.28;12.5 μg/ml,0.31;25 μg/ml,0.48;50 μg/ml,0.54;and 100 μg/ml,0.66 (P<0.05),with the half-effective concentration being around 25 μg/ml.MTT results were confirmed by microscopic analysis.Western blotting and RT-PCR analyses showed that the expressions of caspases 3,6,7,and 9 were significantly decreased in the EuO-treated cells compared with the control (EuOand H2O2-free) (P<0.05),with the half-effective concentration of EuO ranging from 12.5 to 25 μg/ml.We conclude that the ethanol-extracted EuO leaf extracts promoted the growth of MC3T3-E1 cells,and suppressed the H2O2-induced apoptosis in a rat MC3T3-E1 osteogenic cell model,likely due to the inhibition of caspases’ activities.The results indicate that EuO is a potent antioxidant,which may contribute to its many cellular protective functions,including the promotion of bone growth.

Anti-inflammatory effects of Eucommia ulmoides Oliv. male flower extract on lipopolysaccharide-induced inflammation

Background:Eucommia ulmoides Oliv. is a medicinal plant native to China, with its bark (Eucommiae Cortex) traditionally being used for medicinal purposes. Previous research has shown that Eucommia male flowers can exert anti-inflammatory, analgesic, antibacterial, and other pharmacological effects, including immune regulation. This study explored the anti-inflammatory effects of the 70% ethanol extract of male flowers (EF) of E. ulmoides in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and LPS-administered mice.Methods:Cytotoxicity of EF for RAW 264.7 cells was investigated using Cell Counting Kit-8. The production of proinflammatory mediators, nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 was determined using enzyme-linked immunosorbent assays. IL-17, IL-23, and IL-10 mRNA levels were determined using quantitative real-time polymerase chain reaction. Activation of the nuclear factor (NF)-κB pathway in RAW 264.7 cells was investigated via Western blotting. In vivo antiinflammatory effects of EF were studied in an LPS-induced acute inflammation mouse model by analyzing lung tissue histopathology, serum TNF-α and IL-6 levels, and myeloperoxidase (MPO) activity in lung tissue.Results:EF showed no significant cytotoxicity at concentrations from 10 to 60 μg/mL (cell viability > 80%) in the CCK-8 cell viability assay. EF inhibited the RAW 264.7 cell proliferation (EF 60 μg/mL, 120 μg/mL, and 250 μg/mL vs. negative control: 87.31±2.39% vs. 100.00±2.50%, P=0.001;79.01±2.56 vs. 100.00±2.50%, P<0.001;and 64.83±2.50 vs. 100.00±2.50%, P<0.001), suppressed NO (EF 20 μg/mL and 30 μg/mL vs. LPS only, 288.81±38.01 vs. 447.68±19.07 μmol/L, P=0.004;and 158.80±45.14 vs. 447.68±19.07 μmol/L, P<0.001), TNF-α (LPS+EF vs. LPS only, 210.20±13.85 vs. 577.70±5.35 pg/mL, P<0.001), IL-1β (LPS+EF vs. LPS only, 193.30±10.80 vs. 411.03±42.28 pg/mL, P<0.001), and IL-6 (LPS+EF vs. LPS only, 149.67±11.60 vs. 524.80±6.24 pg/mL, P<0.001) secretion, and downregulated the mRNA expression of IL-17 (LPS+EF vs. LPS only, 0.23±0.02 vs. 0.43±0.12, P<0.001), IL-23 (LPS+EF vs. LPS only, 0.29±0.01 vs. 0.42±0.06, P=0.002), and IL-10 (LPS+EF vs. LPS only, 0.30±0.01 vs. 0.47±0.01, P=0.008) in LPS-stimulated RAW 264.7 cells. EF inhibited the LPS-induced NF-κB p65 (LPS+EF 20 μg/mL and 30 μg/mL vs. LPS only: 0.78±0.06 vs. 1.17±0.08, P<0.001;and 0.90±0.06 vs. 1.17±0.08, P=0.002) and inhibitor of kappa B (IκBα) phosphorylation (LPS+EF 20 μg/mL and 30 μg/mL vs. LPS only: 0.25±0.01 vs. 0.63±0.03, P<0.001;and 0.31±0.01 vs. 0.63±0.03, P<0.001), LPS+EF 30 μg/mL inhibited IκB kinase (IKKα/β) phosphorylation (LPS+EF 30 μg/mL vs. LPS only, 1.12±0.14 vs. 1.71±0.25, P=0.002) in RAW 264.7 cells. Furthermore, EF 10 mg/kg and EF 20 mg/kg inhibited lung tissue inflammation in vivo and suppressed the serum TNF-α (LPS+EF 10 mg/kg and 20 mg/kg vs. LPS only, 199.99±186.49 vs. 527.90±263.93 pg/mL, P=0.001;and 260.56±175.83 vs. 527.90±263.93 pg/mL, P=0.005), and IL-6 (LPS+EF 10 mg/kg and 20 mg/kg vs. LPS only, 41.26±30.42 vs. 79.45±14.16 pg/ml, P=0.011;and 42.01±26.26 vs. 79.45±14.16 pg/mL, P=0.012) levels and MPO (LPS+EF 10 mg/kg and 20 mg/kg vs. LPS only, 3.19±1.78 vs. 5.39±1.51 U/g, P=0.004;and 3.32±1.57 vs. 5.39±1.51 U/g, P=0.006) activity in lung tissue.Conclusions:EF could effectively inhibit the expression of inflammatory factors and overactivation of neutrophils. Further investigation is needed to evaluate its potential for anti-inflammation therapy.

杜仲提取物对鲫抗氧化性能的影响

该试验探讨在鲫(Carassius auratus)的饲料中添加不同水平的杜仲提取物对其机体抗氧化性能的影响,确定杜仲(Eucommia ulmoides Oliv.)提取物在鲫养殖中的最适添加水平。试验选择体重约为50g的健康的鲫600尾,按照单因素试验设计,将其均分4个处理,每个处理5个重复,每个重复30尾鲫。对照组鲫投喂基础饲料,试验组鲫投喂添加0.1%、0.2%和0.4%的杜仲提取物的饲料。试验鲫驯养1周,正式试验期6周。试验结束,检测各组鲫机体的抗氧化相关指标。结果显示,投喂0.2%和0.4%的杜仲提取物可以显著降低血清中丙二醛水平(P<0.05),显著提高了过氧化氢酶活力、谷胱甘肽过氧化物酶活力、谷胱甘肽-硫转移酶活力(P<0.05);投喂0.1%、0.2%和0.4%的杜仲提取物显著提高了超氧化物歧化酶活力(P<0.05)。研究结果表明,杜仲提取物可以有效改善鲫机体的抗氧化功能,且杜仲提取物的最适添加量为0.2%。

Plant regeneration from dry mature endosperm cultures of Eucommia ulmoides Oliv.

EUCOMMIA ulmoides is not only a famous medicinal plant,but also a most promising rubber-source plant in the temperate zone.All the individuals present in this species are diploid(2n=2x=34),and the objective of this work is to establish a novel triploid type of it so as to im-prove its overall biomass output or rubber content in the leaf cells by the characteristic cell en-

盐制对杜仲治疗去卵巢大鼠骨质疏松症影响的研究

目的研究杜仲及其盐制品对去卵巢大鼠骨质疏松症的治疗作用,并探讨盐制对其治疗作用的影响。方法 40只SD雌性大鼠分为伪手术组、模型对照组、杜仲生品治疗组和杜仲盐制品治疗组,除假手术组施行假手术外,其余均行手术彻底摘除卵巢,术后4周开始杜仲生品及盐制品水提液灌胃给药(4.0 g/kg·d),实验过程称体质量,连续给药12周后,颈动脉取血,测定血钙(S-Ca)、血磷(S-P)含量,ELISA试剂盒双抗体夹心法测定血清碱性磷酸酶(ALP)、骨钙素(OCN)、雌二醇(E2)含量;动物处死后剥取完整子宫称重,采用HE染色法进行子宫病理学检查,剥离双侧股骨,双能X射线衍射法测定大鼠股骨骨密度。HE染色法观察股骨骨小梁形态和成骨细胞数量,micro-CT扫描观察股骨骨小梁微体系结构。结果杜仲生品及盐制品能有效抑制去卵巢所致的大鼠体重增加,升高血清钙、雌二醇的含量,降低血磷、碱性磷酸酶和骨钙素的含量。增加大鼠股骨骨密度,改善股骨骨小梁微体系结构,增加股骨骨小梁成骨细胞数量,且长期用药对子宫无明显刺激作用。结论杜仲生品及盐制品对于大鼠去卵巢所引起的骨质疏松症有良好治疗作用,且盐制品效果优于生品。

杜仲中桃叶珊瑚苷的提取工艺研究

用正交试验法对杜仲中的活性成分桃叶珊瑚苷的提取工艺进行了优选研究,采用正交表L16(45),以提取时间、提取温度、提取次数、提取液中有机溶剂乙醇含量以及料液比为因素,以桃叶珊瑚苷吸光度为指标进行试验。得到的最佳提取参数为:温度为70℃,时间0.5h,提取次数3次,提取溶剂为80%的乙醇,料液比为1∶9。在此条件下,选取了提取桃叶珊瑚苷的最佳原材料,为杜仲新干皮。

杜仲叶复合保健饮料的研制

利用杜仲叶为主要原料,添加适量新鲜苹果汁及其它辅料,研制出口感优良、风味独特、质量稳定的复合保健饮料。探讨了杜仲叶提取液的脱苦方法,通过正交实验确定了饮料的最佳配方,即:杜仲叶提取液30%,苹果汁20%,白砂糖8%,柠檬酸0.15%,β-环糊精0.2%,VC0.15%,并对饮料进行了急性毒性实验,结果证明此饮料无毒。

杜仲四倍体的诱导与鉴定

以杜仲籽苗为材料，采用秋水仙素溶液处理生长点的诱变方法和茎尖细胞染色体数检测及DNA相对含量测定的倍性鉴定方法，研究了秋水仙素浓度和处理时间的诱变效果。结果表明，除0．05％秋水仙素处理48h外，其它浓度（0．1％，0．2％，0．4％）均以处理12h效果最好，变异株率在43．3％～63．3％，最佳处理组合为0．1％秋水仙素处理生长点12h，变异株率和四倍体株率分别为63．3％和36．7％。在获得的148个变异株中，染色体计数表明47株为四倍体（2n=4x=68），占31．76％；染色体计数结果进一步被流式细胞仪分析验证。四倍体植株表现出多倍体特征，叶片变厚，叶面积增大，叶形指数减小，保卫细胞增大，单位面积气孔数减少，叶绿体数明显增多。

杜仲叶对草鱼生长、血清非特异性免疫指标和肉质影响的初步研究

为考察杜仲叶对草鱼生长、血清非特异性免疫指标和肌肉品质的影响,在基础饲料(对照组)中分别添加2%4、%杜仲叶粉或0.1%0、.15%杜仲叶提取物,饲养平均体重(425.8±37.6)g的草鱼。经60d饲养,各组草鱼增重率分别为84.3%、92.2%、96.7%、88.2%、91.7%,饲料系数分别为2.01、1.87、1.82、2.03、1.91;与对照组相比,添加4%杜仲叶粉显著提高了草鱼增重率,降低了饲料系数(P<0.05),添加2%杜仲叶粉、0.15%杜仲叶提取物有提高草鱼增重率趋势(P<0.10)。对血清非特异性免疫指标的测定表明,饲料中添加4%杜仲叶粉,0.1%、0.15%杜仲叶提取物,均可极显著提高SOD活性(P<0.01),各组溶菌酶和碱性磷酸酶活性无显著差异(P>0.05)。对肌肉成分的分析表明,各组间肌肉水分、灰分含量无显著差异(P>0.05),但添加0.15%杜仲叶提取物,可显著提高肌肉蛋白、谷氨酸、丝氨酸、丙氨酸、苯丙氨酸、组氨酸、脯氨酸、呈味氨基酸及总氨基酸含量(P<0.05);添加4%杜仲叶粉、0.1%、0.15%杜仲叶提取物均可提高肌肉羟脯氨酸、胶原蛋白含量。对肌肉物理性能指标的分析表明,添加杜仲叶粉或叶提取物均可降低肌纤维直径(P<0.05),其中,添加4%杜仲叶粉、0.1%、0.15%杜仲叶提取物还提高了肌原纤维耐折力(P<0.05),添加2%、4%杜仲叶粉,0.15%杜仲叶提取物降低了肌肉失水率(P<0.05)。上述研究表明,饲料中添加杜仲叶(叶粉或叶提取物)可改善养殖草鱼生长性能、血清非特异性免疫功能和肌肉品质。

杜仲内生真菌的分离鉴定及抗菌活性研究

采用组织分离法从秦岭药用植物杜仲茎部共分离到内生真菌38株,经形态学鉴定分属于9属,其中青霉属为杜仲内生真菌中的优势菌群,拟青霉属、交链孢属和无孢群属为常见属。对分离得到的杜仲内生真菌发酵液进行了抑菌实验,其中61%的菌株对测试细菌具有抗菌活性,表明杜仲内生真菌的抗菌活性十分普遍;34%的菌株对烟草赤星病菌、苹果炭疽病菌及辣椒炭疽病菌等多种植物病原菌具有较强的抑制活性,表明杜仲内生真菌在植物病害的生物防治领域具有进一步开发的价值。

杜仲的化学成分及药理作用研究进展

综述了杜仲化学成分及同类化合物的药理作用研究成果。通过与同类化合物的药理作用研究比较,为杜仲活性功能的物质基础研究以及新型活性功能的开发提供参考信息。