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%。

Comprehensive Assessment of the Safety of Eucommia ulmoides Leaf Extract for Consumption as a Traditional Chinese Health Food

To ensure the export quality of Eucommia ulmoides leaf extract(ELE)and facilitate E.ulmoides leaf inclusion in the directory of traditional Chinese health foods,an overall safety assessment of ELE was performed,including genotoxicity and long-term toxicity,according to the national food safety standards of China.No variations in the reverse mutation number of the nominal bacterial strains were observed under ELE treatment in comparison with the solvent control.Additionally,the micronucleus rates of in vivo mammalian erythrocytes and in vitro mammalian cells under ELE treatment were equivalent to or significantly lower than those of the solvent control.The fold change in the trifluorothymidine resistance mutation frequency of the thymidine kinase gene under ELE treatment was less than three times in comparison with the solvent control,suggesting that ELE did not cause genotoxicity.Moreover,animal experiments showed that the growth performance of rats under ELE treatment was enhanced because the body weights of rats increased.No oxidative injury or inflammatory responses were induced and no histopathological lesions of tissues were detected under ELE treatment.In addition,plasma triglycerides and low-density lipoprotein cholesterol levels significantly decreased,and plasma high-density lipoprotein cholesterol levels significantly increased with ELE treatment,suggesting that ELE was health-promoting.Furthermore,moderate to excellent antimicrobial activities,a favorable anticancer capacity,and superior antioxidant abilities of ELE were found,implying ELE possesses good bioactivities.Therefore,we affirmed ELE is safe to consume as a traditional Chinese health food.

中药杜仲皮总黄酮提取工艺及抗氧化活性研究

优化杜仲(Eucommia ulmoides Oliv.)皮总黄酮提取工艺,建立了总黄酮含量测定方法,并对其体外抗氧化活性进行研究。采用回流提取法提取杜仲皮总黄酮,通过单因素试验,考察了提取方法、溶剂浓度、溶剂体积、提取时间对总黄酮含量的影响,采用正交试验优化提取工艺,筛选出杜仲皮总黄酮的最佳提取工艺;测定总抗氧化能力、羟基自由基清除自由基能力判断杜仲皮总黄酮的体外抗氧化活性。结果表明,杜仲皮总黄酮的最佳提取工艺为50%乙醇、料液比1∶40 g/mL、回流提取时间1.5 h;总黄酮具有较强的还原能力,浓度为3.0 mg/mL时其总抗氧化能力为132.2 U/mL,总抗氧化能力随着总黄酮浓度的增加而增强,相同溶液浓度下较维生素C强;总黄酮对羟基自由基有较强的清除能力,溶液浓度为3.0 mg/mL时,羟基自由基清除率为90%,清除率随总黄酮浓度的增加而增大,最后趋于平稳。该方法能有效测定杜仲皮总黄酮的含量,提取的总黄酮具有良好的体外抗氧化活性,精密度、准确性、稳定性、重复性较好,为该药材的质量标准研究提供参考。

Ectopic Overexpression of EuCHIT30.7 Improves Nicotiana tabacum Resistance to Powdery Mildew

Various strains of powdery mildew(PM),a notorious plant fungal disease,are prevalent and pose a significant threat to plant health.To control PM,transgenic technology can be used to cultivate more resistant plant varieties.In the present study,we utilized the rapid amplification of cDNA ends(RACE)technique to clone the full-length cDNA sequence of the EuCHIT30.7 gene to explore plant genes with disease resistance functions.Bioinformatics analysis revealed that this gene belongs to the GH18 family and is classified as a class III chitinase.The EuCHIT30.7 gene is expressed throughout the Eucommia ulmoides plant,with the most abundant expression in male flowers.Subcellular localization analysis indicated that the protein encoded by this gene was detected within both the cell membrane and cytoplasm.Upon PM inoculation,overexpression of EuCHIT30.7 in tobacco plants led to a significantly reduced relative lesion area and a decreased spore count compared to both wild-type and empty vector control plants.Activities of the protective enzymes,namely,peroxidase(POD),superoxide dismutase(SOD),catalase(CAT),and phenylalaninammo-nialyase(PAL),in tobacco plants overexpressing EuCHIT30.7 were significantly greater than those in wild-type and empty vector tobacco plants.Furthermore,the rate of increase in malondialdehyde(MDA)content was significantly lower in tobacco plants expressing EuCHIT30.7 compared to control tobacco plants.In EuCHIT30.7 transgenic tobacco,the expression of pathogen-related protein genes,namely,PR2,PR5,PR1a,PDF1.2,and MLP423,along with the tobacco PM negative regulatory gene,MLO2,were significantly higher compared to control tobacco plants.These findings suggested that EuCHIT30.7 significantly enhances the resistance of tobacco to PM.

Cloning and Function Identification of a Phytoene Desaturase Gene from Eucommia ulmoides

The phytoene desaturase(PDS)encodes a crucial enzyme in the carotenoid biosynthesis pathway.Silencing or inhibiting PDS expression leads to the appearance of mottled,chlorosis,or albino leaves.In this study,the CDS sequence of EuPDS(Eucommia ulmoides Phytoene Desaturase)was first cloned and then PDS was silenced in Nicotiana benthamiana.Result showed the expression level of EuPDS in leaves was higher than that in the roots and stems.In N.benthamiana leaves,which were treated by Agrobacterium for 24 h,photo-bleaching was shown on the fresh leaves one week after injection and the transcript level of PDS was down-regulated during the period of emersion.This suggested that EuPDS could silence PDS of N.benthamiana,so as to cause the phenotype of leaf whitening.PDS is the main reporter gene involved in virus-induced gene silencing(VIGS).This study offered molecular evidence for identifying PDS gene involved in Carotenoid’s biosynthesis pathway and the regulation networks in E.ulmides.It also laid a useful foundation for study on leaf discoloration mechanism of other woody plants.

Overexpression ofβ-1,4-Glucanase Gene EuEG1 Improves Micrografting of Eucommia ulmoides

Adventitious root formation poses a major constraint on the tissue culture and genetic transformation of Eucommia ulmoides.Micrografting offers a new method for the transplantation of genetic transformation,and its success depends on the formation of graft unions.This study used transgenic rootless test-tube seedlings as scions and seedlings from seed as rootstocks during micrografting to avoid the rooting issues that occur during tissue culture and to investigate the role of the EuEG1 gene in the graft healing process.We found that the EuEG1 gene is a vital regulator of graft,and its overexpression contributes to the survival of Eucommia ulmoides micrografting.The EuEG1 gene transgenic plants(TP)used as scions for micrografting presented a significantly higher survival rate than the wild type(WT)and empty vector(EV)regenerated scions.During the grafting healing process,the expression of the EuEG1 gene was higher during the period of callus proliferation,suggesting that the EuEG1 gene was involved in the graft healing process.Histological observation revealed that more calluses tissue appeared at the junction of transgenic scions,and the connection with the rootstock was stronger,which benefits wound healing.These results provide new insights into Eucommia ulmoides micrografting and indicate that the EuEG1 gene can promote wound healing and improve the micrografting survival rate.

Role and Effects of Eucommia ulmoides Leaves in the Growth Process of Livestock and Poultry

Substitutes of feed antibiotics have been a key research topic in the new stage of animal husbandry.Chinese veterinary medicine refers to decoction pieces and their preparations processed from natural plants,animals,and minerals and used for animal disease prevention and improvement of animal production performance under the guidance of Chinese veterinary pharmacy theory.Eucommia ulmoides leaf extract has many active functional components such as chlorogenic acid,E.ulmoides polysaccharides and flavonoids,which have many biological properties such as antibacterial,antioxidant,immune-regulation,sugar ester-and bone metabolism-regulation effects.This paper explored active ingredients and biological properties of E.ulmoides leaf extract,as well as its role and effects in livestock and poultry breeding,providing a scientific basis for the use of E.ulmoides leaves in livestock and poultry breeding.

SFE-CO_2法与SD法提取杜仲叶挥发油成分的比较

分别采用超临界CO2流体萃取法(SFE-CO2法)与水蒸气蒸馏法(SD法)从杜仲(Eucommia ulmoids Oliv.)叶中提取挥发油,并结合气相色谱-质谱仪(GC-MS)对其进行检测。从SFE-CO2法提取的杜仲叶挥发油中共鉴定出59种成分,占挥发油总量的82.57%;从SD法提取的杜仲叶挥发油中共鉴定出39种成分,占挥发油总量的84.48%。两种提取方法得到的挥发油组分及其含量差异较大。SFE-CO2法比SD法能更真实、全面的反映杜仲叶药材的挥发油成分。

杜仲皮和叶中儿茶素含量差异的研究

[目的]研究杜仲皮和叶中儿茶素含量的差异,为杜仲以叶代皮入药提供依据。[方法]选取3个产地的杜仲皮和叶样品进行提取,并采用反相高效液相色谱法测定儿茶素的含量,色谱柱为Agilent HC-C18(4.6 mm×250 mm,5μm);流动相为乙腈-浓度0.4%磷酸溶液(13∶87,V/V);流速为1.0 ml/min,检测波长280 nm。[结果]所取3个产地的同株杜仲的皮和叶中儿茶素的含量不同,其中河南信阳、四川绵阳和贵州遵义产的杜仲皮中儿茶素的含量分别为1.919、1.279和1.167 mg/g,所产的杜仲叶中儿茶素的含量分别为49.34、35.75和33.72 mg/g;同株杜仲叶中儿茶素含量明显高于杜仲皮。[结论]研究所采用分析方法简便、快速,重现性良好,适用于杜仲不同部位儿茶素含量的测定。测定结果表明杜仲叶资源值得开发利用。

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。

HPLC法同时测定水解前后杜仲叶黄酮苷元的含量

建立快速检测杜仲中两种黄酮苷元槲皮素与山奈酚的高效液相色谱方法，并测定水解前后杜仲叶与杜仲素黄酮苷元的含量。采用 HPLC 法同时测定黄酮苷元，色谱柱为 Diamonsil C18（4.6 mm ×250 mm，5μm），以甲醇-0.2％磷酸（63：37）为流动相，检测波长为370 nm，流速1.0 mL/ min，柱温为30℃。槲皮素和山奈酚分别在0.5~80μg/ mL 和0.5~18μg/ mL 内线性关系良好，该检测方法能迅速、准确检测水解前后杜仲叶和杜仲素中槲皮素和山奈酚的含量；水解工艺可明显提高杜仲黄酮苷元含量。

杜仲穴盘育苗基质的研究

以泥炭、珍珠岩、蛭石等常用育苗基质为材料,对杜仲(Eucommia ulmoides Oliver)进行了不同基质配方的育苗效果试验。结果表明,以泥炭∶珍珠岩∶蛭石=3∶0∶2的基质配方育苗效果好,比较适合杜仲穴盘育苗方式。

多效唑对杜仲穴盘育苗质量的影响

采用穴盘育苗方法,研究了不同浓度(30、60、120、240、480、960 mg/L)多效唑处理对杜仲(Eucommia ulmoides)幼苗质量的影响。结果表明,杜仲育苗过程中叶片喷施多效唑浓度以240 mg/L处理效果最好,与对照相比,株高降低了22.6%,根冠比增加了56.4%,壮苗指数增加了42.5%。

HPLC测定不同树龄杜仲叶中黄酮类成分的含量

[目的]建立高效液相色谱同时测定不同树龄杜仲叶中绿原酸、芦丁、槲皮素和山萘酚含量的方法。[方法]采用Shim-pack VP-ODS(4.6×150 mm,5μm)色谱柱,以甲醇-0.4%磷酸水溶液为流动相进行梯度洗脱,流量1.0 ml/min,用二极管阵列检测器扫描检测,以保留时间结合待测物质的紫外吸收光谱进行定性分析。采用外标法进行定量分析,检测波长为360 nm,柱箱温度为40℃。[结果]绿原酸的线性范围为4.63～92.65μg/ml,r=0.999 8,回收率为98%～101.5%;芦丁的线性范围为0.714～14.285μg/ml,r=0.999 9,回收率为96%～98.4%;槲皮素的线性范围为0.101～2.066μg/ml,r=0.997 0,回收率为94%～101%;山萘酚的线性范围为0.043～0.886μg/ml,r=0.998 0,回收率92%～96%。[结论]对不同树龄杜仲叶中4种有效成分含量用方差分析进行多重比较,从绿原酸含量方面考虑,三年树龄杜仲叶采收最佳。

杜仲组织培养的研究

为了筛选出杜仲(Eucommia ulmoides)组织培养的最优培养条件,主要从外植体的选择、基本培养基、植物激素、光照条件、其他理化条件等方面对杜仲愈伤组织的诱导、增殖及分化培养的影响进行了综述,分析了杜仲组织培养过程中常见的问题并提出了解决方案,以期为杜仲快繁体系的建立提供理论依据。

杜仲内生真菌ER12次生代谢产物研究

[目的]对杜仲内生真菌ER12次生代谢产物进行研究。[方法]采用色谱技术进行分离纯化,并采用理化性质及波谱分析技术确定化合物的结构。[结果]从杜仲内生真菌ER12次生代谢产物中分离并鉴定了4个化合物,分别是正丁基-β-D-吡喃果糖苷(1)、甘露醇(2)、啤酒甾醇(3)和麦角甾醇(4)。[结论]该方法明确了杜仲内生真菌ER12的次生代谢产物。

Ca^(2+)对杜仲愈伤组织诱导及增殖效应的影响研究

以杜仲(Eucommia ulmoides)幼茎为外植体,进行杜仲愈伤组织诱导、增殖实验。在培养基中添加不同浓度的Ca2+进行筛选实验,以确定诱导、增殖培养基中的最佳Ca2+浓度。结果表明适度增加Ca2+浓度,可促进杜仲愈伤组织的诱导和增殖,并能改善愈伤组织的生长状态,增加愈伤组织增殖生长量。但若Ca2+浓度超过12.0 mmol/L,促进效果减弱。

杜仲活性成分四氢呋喃骈四氢呋喃型木脂素的快速分析

杜仲(eucommia ulmoides Oliv)具有补肝肾、强筋骨、安胎和降压等功效.其中四氢呋喃骈四氢呋喃型木脂素松脂素二葡萄糖苷是其主要的降压有效成分;丁香脂素二葡萄糖苷在治疗淋巴细胞白血病中有较好的活性[1].……

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.

杜仲叶中绿原酸提取工艺的优化

分别以水和乙醇溶液为溶剂从杜仲(Ecommia ulmoides Oliv.)叶中提取绿原酸,采用高效液相色谱法测定提取物中绿原酸的含量并计算提取率,设计正交试验优化提取工艺。结果表明,优化的醇提法工艺条件为料液比1∶15(m∶V,g/mL)、体积分数30%的乙醇作溶剂、提取时间2.0 h、提取次数2次;优化的水提法工艺条件为料水质量比1∶10、提取温度60℃、提取时间1.5 h、提取次数2次。