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.

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.

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.

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.

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.

南方红壤区杜仲(Eucommia ulmoides)树干液流动态

采用根据热平衡原理设计的热扩散探针(Thermal dissipation sap flow velocity probe,TDP),于2004年7月到10月对南方红壤区的杜仲人工林的树干液流进行连续监测,结合所测定的相关因子,分析杜仲液流的变化规律及其与各因子的关系。结果表明,在不同月份,杜仲液流速率的日变化规律基本一致,呈单峰曲线,但是树干液流在启动时间、峰值出现时间上存在差异;从7月到10月份,杜仲树干液流速率逐渐增大,10月份液流速率达到1.818 g/(cm2.h);杜仲液流速率与太阳辐射、空气相对湿度、温度、风速等气象因子相关性显著,但是太阳辐射和空气相对湿度是影响杜仲液流的重要因子,其中7月到9月份,太阳辐射是主要影响因子,而10月份,空气相对湿度占主导地位;树干液流与胸径、冠层厚度以及胸径平方与树高之积呈现出显著的相关性。

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.

Determining the in vitro Anti-Aging Effect of the Characteristic Components from Eucommia ulmoides

To evaluate the potential anti-aging ability of Eucommia ulmoides,four characteristic components(chlorogenic acid,geniposidic acid,aucubin,quercetin)were selected to assess their effects on H_(2)O_(2)-induced oxidative damage model of human umbilical vein endothelial cell(HUVEC).Oxidative damage indexes,inflammatory factors,cell cycle,cell apoptosis,cell senescence,and their related proteins were analyzed by methyl thiazolyl tetrazolium(MTT)assay,enzyme-linked immunosorbent assay(ELISA),propidium iodide(PI)staining,annexin V-FITC/PI double staining,SAβ-galactosidase staining,and western blotting(WB).The results showed that H_(2)O_(2)-induced cell growth inhibition rate decreased as supplementation with characteristic components when compared to H_(2)O_(2) group.Meanwhile,the contents of antioxidant indexes(reactive oxygen species,lactate dehydrogenase,molondialdehyde,superoxide dismutase,glutathione),inflammatory factors(nuclear factor kappa-B,intercellular cell adhesion molecule-1,vascular cell adhesion protein 1),and functional factors(NO,Endothelin-1)in characteristic components treated groups improved if comparison with H_(2)O_(2) group,suggesting the characteristic components of E.ulmoides could alleviate H_(2)O_(2)-induced oxidative damage.Moreover,cell cycle,cell apoptosis,cell senescence,and their related proteins under characteristic components treatment exhibited a better effect than under H_(2)O_(2) treatment,implying the characteristic components could participate in anti-aging via multiple pathways.These results manifested that the characteristic components of E.ulmoides posses the capacity of anti-aging,which provided a basis for investigating the anti-aging ability of E.ulmoides itself.

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-too/des by NaC1 extract,gel filtration and reverse phase high performanee liquid ehromatography.Themolecular mass of EAFP3 is 4157.3 Da,and its partial amino acid sequenee is-.LYQQLIAGITLNK.-.EAFP3 exerts an inhibitory activity against Candida albicans in vitro and the drug concentration requiredfor 50% growth inhibition(IC50)is 31.25μg/mL.

Transcriptomic and Physiological Analyses of Pigment Accumulation in Eucommia ulmoides‘Hongye’

Eucommia ulmoides‘Hongye’is a new ornamental variety of E.ulmoides with excellent red or purple foliage.We found that E.ulmoides‘Hongye’exhibited a gradual change from green to red colour under light conditions.However,the colouring mechanism in the leaves of E.ulmoides‘Hongye’remains unclear.In this study,we compared the pigment content and leaf colour index of E.ulmoides‘Hongye’at five stages with those of E.ulmoides‘Xiaoye’,which was used as the control variety.The transcriptome sequencing data of the first-period(H1,green)and fifth-period(H5,red)leaves were also analysed and compared.The corresponding gene regulation in anthocyanin-related metabolic pathways was then analysed.Physiological results indicated that the contents of flavonoids and anthocyanins in red leaves(H5)were significantly higher than those in green leaves(H1),whereas the chlorophyll content in red leaves(H5)was lower than that in green leaves(H1).Moreover,the carotenoid content did not significantly differ between the two varieties.A transcriptome analysis identified 4240 differentially expressed genes(DEGs),and 20 of these genes were found to be involved in flavonoid and anthocyanin biosynthesis pathways.The results provide a reference for further study of the leaf colouration mechanism in E.ulmoides.

杜仲Eucommia ulmoides的核型分析

以萌发种子的根尖为材料,对兰州地区引种的杜仲进行了体细胞染色体核型分析。结果显示:杜仲染色体较小,数目均为34条,与前人的研究结果一致;杜仲体细胞染色体核型有2类,第1类为2n=34=20m(2SAT)+10sm+4st,第2类为2n=34=20m(2SAT)+12sm+2st,在2类核型中均见到了随体染色体,随体均位于中部着丝点染色体的短臂上。2类核型中最长染色体与最短染色体的比值分别小于2,臂比值大于2∶1的染色体的比例在1%～50%之间,按照stebbins的核型分类标准应属2A核型,与前人的核型分析结果差异较大。

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。

不同良种杜仲叶主要营养成分的动态变化与综合评价

[目的]杜仲Eucommia ulmoides叶在医疗、保健、食品及无抗养殖等领域有广泛用途。通过比较不同良种杜仲叶主要营养成分的差异及动态变化,筛选优质叶用杜仲良种资源,为杜仲叶的高效利用提供参考。[方法]分别利用HPLC法和全自动氨基酸分析仪测定13个良种不同生长期杜仲叶9种重要活性成分和15种氨基酸组分的含量,基于主成分分析和隶属函数法开展不同良种杜仲叶的营养评价。[结果]与槲皮素、桃叶珊瑚苷和京尼平苷酸相比,原儿茶酸、芦丁、绿原酸、紫云英苷、山奈酚以及松脂素二葡萄糖苷的含量在不同良种杜仲叶之间差异较大;不同良种同一活性成分含量的生长动态变化趋势基本一致。13个良种杜仲叶必需氨基酸与总氨基酸的比值(EAA/TAA)为0.40~0.45,必需氨基酸与非必需氨基酸的比值(EAA/NEAA)为0.66~0.82;天冬氨酸、苏氨酸、丝氨酸等5种氨基酸的动态变化趋势较为一致。总氨基酸、必需氨基酸、亮氨酸等9种组分的含量可作为杜仲叶营养价值的评判指标。[结论]13个良种杜仲叶的氨基酸组成和比例均达到理想蛋白质模式的标准。‘华仲10号’‘华仲24号’‘华仲13号’‘华仲30号’‘华仲12号’是优质的叶用杜仲良种资源,在规模化提取和利用杜仲叶黄酮类时应优先选用‘华仲12号’‘华仲24号’良种。

杜仲改善大鼠产后抑郁的作用机制研究

目的 探讨杜仲改善大鼠产后抑郁的作用机制。方法 受孕雌鼠随机分为正常组、产后抑郁组和杜仲低、高剂量组(1.34、2.68 g/kg,以生药计),每组10只。除正常组外其余各组大鼠运用孕期旁观电击法建立产后抑郁大鼠模型;造模的同时,给药组大鼠灌胃相应药物,正常组及产后抑郁组大鼠灌胃生理盐水,持续21 d。观察各组大鼠实验期间的一般情况,通过旷场实验、Morris水迷宫实验和糖水偏好实验进行行为学评价,检测各组大鼠血清中皮质酮(CORT)、下丘脑中促肾上腺皮质激素释放因子(CRF)和尿皮质素(UCN)、垂体中促肾上腺皮质激素(ACTH)水平,以及海马组织中CRF受体1(CRFR1)、CRFR2及电压依赖性阴离子通道1(VDAC1)蛋白表达水平,海马组织凋亡细胞比例及JC-1高电位细胞比例,并观察海马组织形态。结果 与产后抑郁组比较,杜仲高剂量组大鼠食欲、精神、毛色均有所改善,体重有所增加;垂直运动、水平运动、自我梳理得分均显著升高(P<0.05);第2~4天逃避潜伏期均显著缩短;穿越平台次数显著增加,每次穿越时间显著延长(P<0.05);孕20 d及产后30 d糖水消耗比率显著升高(P<0.05);CRF、UCN、ACTH、CORT水平,噬仔率,CRFR2、VDAC1蛋白表达水平,海马组织凋亡细胞比例均显著降低(P<0.05);JC-1高电位细胞比例显著升高(P<0.05);神经元细胞周围水肿现象明显改善。结论 杜仲可能通过抑制下丘脑-垂体-肾上腺轴的过度激活,降低CRFR2的表达水平,进而抑制VDAC1的表达,并减少神经元细胞的凋亡,从而改善产后抑郁症状。

超声处理对杜仲籽仁蛋白结构及理化特性的影响

旨在为杜仲籽仁蛋白的开发与应用提供参考,以杜仲籽仁粕为原料,采用碱溶酸沉法提取蛋白,采用500 W、35 kHz超声波对杜仲籽仁蛋白处理不同的时间(0、10、20、30 min和40 min),探究超声处理对杜仲籽仁蛋白微观结构、理化特性(粒径、Zeta-电位、内源荧光光谱、溶解性、持水性、持油性、起泡性、起泡稳定性、乳化性、乳化稳定性)的影响。结果表明:超声处理可改变杜仲籽仁蛋白的微观结构、粒径、Zeta-电位和荧光强度;超声处理20 min时,杜仲籽仁蛋白溶解度达到75.35%;超声处理10 min时,杜仲籽仁蛋白乳化性最大,达到72.12 m^(2)/g;超声处理40 min时,杜仲籽仁蛋白持水性、持油性、起泡性和起泡稳定性最大,分别达到4.32 g/g、3.46 g/g、23.12%和21.12%。综上,超声处理可改善杜仲籽仁蛋白的持水性、持油性、起泡性、乳化性等功能特性。

杜仲叶 红枣 枸杞复合饮料工艺配方的研究

以杜仲叶、红枣、枸杞为原料,辅以柠檬酸和木糖醇,优化和确定复合饮料工艺配方。以感官评分为考查指标,利用单因素试验和正交试验对影响复合饮料的杜仲叶添加量、红枣添加量、枸杞添加量、柠檬酸添加量、木糖醇添加量配方组合进行优化。结果表明,杜仲叶浸提液添加量15%,枸杞浸提液添加量20%,红枣浸提液添加量15%,柠檬酸添加量0.1%,木糖醇添加量9%为复合饮料的最佳配方。此复合配方饮料酸甜适中风味最佳,生产方法简单可行。

中药复方治疗原发性骨质疏松用药规律及作用机制分析

背景:中药复方治疗原发性骨质疏松有着较久的历史,并且疗效确切,但其用药规律及作用机制尚不明确。目的:利用数据挖掘与网络药理学的方法学,探究现代中医家治疗原发性骨质疏松的用药规律、分子层面的作用机制并一定程度的加以验证。方法:将中国知网、万方、维普、PubMed等数据库收录的相关文献作为数据来源,使用Microsoft EXCEL 2019、IBM SPSS 25.0等软件统计和提取相关资料。将数据统计得到的高频药物进行关联规则分析和聚类分析,综合两者结果得到中药复方治疗原发性骨质疏松的核心药物组合,利用网络药理学阐释此组合发挥治疗作用的机制并借助分子对接加以验证。结果与结论:最终纳入151篇文献,筛选出方剂207首,涉及中药285味。①通过上述两种分析综合得到重要药物组合10组,其中置信度和提升度最高的核心药物组合为“骨碎补-杜仲-当归”,其组合治疗原发性骨质疏松的关键成分为槲皮素、山奈酚、柚皮素等,核心靶点为SRC原癌基因(SRC proto-oncogene)、磷脂酰肌醇-3-激酶调节亚单位1(PIK3R1)、RELA原癌基因(RELA)等,主要通路为癌症信号通路、JAK-STAT信号通路、VEGF信号通路、核因子κB信号通路等。②将关键活性成分与核心靶点进行分子对接,两者展示出较好的结合性。③中药复方治疗原发性骨质疏松核心药物组合可利用其多种活性成分,通过多条信号通路、作用于多个靶点发挥疗效,可为后续治疗原发性骨质疏松新药的研发提供理论基础。

基于电子鼻和HS-SPME-GC-MS分析蒸汽爆破对杜仲嫩叶和成熟叶茶香气特征的影响

为探究蒸汽爆破(汽爆)辅助加工制成杜仲叶茶的挥发性成分差异和主要呈香特征,以杜仲‘华仲8号’嫩叶和成熟叶为对象,采用汽爆技术对杜仲叶进行处理,基于电子鼻和顶空固相微萃取-气相色谱-质谱(headspace solid-phase microextraction combined with gas chromatography-mass spectrometry,HS-SPME-GC-MS)联用法解析汽爆对两种杜仲茶香气特征的影响。主成分分析和线性判别分析对电子鼻数据模型拟合度高,杜仲嫩叶和成熟叶茶汽爆前后香气特征均存在显著差异。HS-SPME-GC-MS检测共鉴定出177种挥发性成分,通过正交偏最小二乘判别分析(变量重要性投影值≥1)以及Kruskal-WallisH检验(P<0.05)进一步筛选出24种香气物质,发现汽爆前嫩叶茶关键香气物质为二氢猕猴桃内酯。汽爆后嫩叶茶关键呈香物质在此基础上增加了壬醛、苯甲醛和苯乙醛,呈现柑橘香、花香、焦糖香、苦杏仁味、坚果香、玫瑰香和巧克力香。汽爆前成熟叶茶无关键香气物质,汽爆后成熟叶茶关键香气物质为二氢猕猴桃内酯和壬醛,呈现甜桃香、木香、柑橘香、花香和焦糖香。研究结果可为开发杜仲叶茶饮品提供参考依据。