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 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.

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-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.

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。

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%。

杜仲水提物上调Nur77表达促进骨髓间充质干细胞增殖和成骨分化

背景:骨髓间充质干细胞作为成骨细胞的前体细胞,探索其分化机制对于预防和治疗骨质疏松症具有重大意义。杜仲能够诱导骨髓间充质干细胞成骨分化,但是杜仲对骨髓间充质干细胞增殖、分化以及Nur77/MDM2/p53通路的影响尚不明确。目的:探究杜仲水提物对骨髓间充质干细胞增殖和分化的影响并探讨其作用机制。方法:杜仲水提物处理人源和鼠源骨髓间充质干细胞,采用CCK-8法和Annexin V-FITC/PI法分别检测细胞增殖和凋亡情况,Western blot检测增殖、凋亡相关蛋白的表达,Nur77及其下游MDM2、p53相关蛋白的表达及泛素化前后的蛋白水平。成脂、成骨定向诱导分化后,Western blot检测脂肪细胞和成骨细胞的标志蛋白表达来评估其分化程度。转染Nur77 siRNA进入10 mg/mL杜仲水提物处理的骨髓间充质干细胞,检测细胞增殖、凋亡和分化情况。最后,敲低Nur77之后,使用p53抑制剂Pifithrin-α处理骨髓间充质干细胞,检测细胞增殖、凋亡和分化情况以明确其作用机制。结果与结论:①杜仲水提物可促进2种骨髓间充质干细胞的增殖和成骨分化,抑制凋亡和成脂分化;②杜仲水提物可上调Nur77、MDM2蛋白表达,促进p53蛋白泛素化;敲低Nur77基因则抑制MDM2蛋白表达及p53蛋白泛素化,促进p53蛋白表达,抑制2种骨髓间充质干细胞增殖和成骨分化;③p53抑制剂可逆转Nur77 siRNA的作用,促进2种骨髓间充质干细胞的增殖和成骨分化,抑制凋亡和成脂分化;④结果表明,杜仲水提物可以通过Nur77/MDM2/p53途径促进2种骨髓间充质干细胞的增殖和成骨分化,为杜仲在骨相关疾病方面的研究提供理论参考。

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.

全杜仲胶囊对自发性高血压大鼠肠壁微血流灌注的影响

目的 探讨全杜仲胶囊(QDZJN)和松脂醇二葡萄糖苷(PDG)对自发性高血压大鼠(SHRs)肠壁微循环血流灌注及其频谱特征的影响。方法 将SHRs随机分为SHR,SHR+QDZJN,SHR+PDG(L),SHR+PDG (M)和SHR+PDG (H)组。对照组为Wistar-kyoto(WKY)鼠。各组分别接受QDZJN或纯净水(对照组和SHR组)灌胃2个月。手术暴露大鼠小肠,激光多普勒血流探测仪(LDF)和血流成像仪(LDPI)检测并比较各组大鼠肠壁微血流灌注水平。小波分析比较各组肠壁内皮源性频段幅值。结果 与对照组相比,SHRs肠壁血流灌注水平显著降低(P<0.001)。QDZJN和各剂量PDG治疗2个月后,SHRs肠壁血流灌注水平显著升高(P<0.001)。SHRs组肠壁微血流灌注一氧化氮(NO)依赖和非依赖性内皮细胞源频段振幅均显著低于对照组(P<0.01)。QDZJN和PDG治疗可显著提高内皮细胞源频段幅值(NO依赖:P<0.0001;NO非依赖:P<0.01)。结论 QDZJN和PDG治疗2个月可显著改善SHRs肠壁的微血流动力学和内皮功能障碍。

杜仲对胶原蛋白代谢的影响及其在腹股沟疝筋膜薄弱大鼠中的应用

目的:观察杜仲提取物(Eucommia ulmoides Oliver extract, EUOE)对大鼠腹股沟疝筋膜薄弱的影响,并初步探索其对人皮肤成纤维细胞(human skin fibroblasts, HSFs)胶原蛋白代谢的作用机制。方法:将32只SD大鼠随机分为正常对照组、模型组、低剂量(1 g/kg)EUOE治疗(low-dose EUOE, EUOE-L)组和高剂量(2 g/kg)EUOE治疗(high-dose EUOE, EUOE-H)组,每组8只。建立大鼠腹股沟区筋膜薄弱模型,给予EUOE干预后,通过生物力学实验检测筋膜抗张强度并采用HE染色观察筋膜组织学结构。采用0~2 g/L EUOE体外孵育HSFs,CCK-8实验检测其对细胞活力的影响;随后通过细胞黏附实验、划痕实验、Transwell细胞迁移实验及ELISA实验观察EUOE(1 g/L)对体外培养HSFs增殖、黏附、迁移及Ⅰ型胶原蛋白(collagen type Ⅰ, COL Ⅰ)分泌的影响,同时利用Western blot实验初步分析HSFs及大鼠筋膜组织中COL Ⅰ、COL Ⅲ和基质金属蛋白酶1(matrix metalloproteinase-1, MMP-1)的表达情况,以及RhoA/ROCK信号通路的活性。结果:体内动物实验结果显示,与模型组相比,给予EUOE治疗可显著增强筋膜的抗张强度(P<0.05),并改善筋膜的组织形态。体外细胞实验的结果显示,EUOE对人皮肤成纤维细胞无显著毒性,同时1 g/L的EUOE孵育48 h可显著增强细胞活力并促进细胞黏附及迁移(P<0.05)。此外,EUOE还可促进HSFs分泌COL I(P<0.05),上调HSFs及大鼠筋膜组织中COL Ⅰ和COL Ⅲ的表达,激活RhoA/ROCK信号通路,抑制MMP-1的表达(P<0.05)。结论:EUOE可显著增强腹股沟疝筋膜薄弱模型大鼠筋膜的抗张强度,该作用可能与其激活RhoA/ROCK信号通路,调节胶原蛋白的代谢有关。

杜仲叶粉对产蛋后期蛋鸡生产性能、蛋品质、血清生化指标及蛋黄胆固醇的影响

试验旨在研究杜仲叶对海兰褐蛋鸡生产性能、蛋品质、血清生化指标及蛋黄胆固醇的影响。选取720只327日龄海兰褐蛋鸡随机分为4组,每组5个重复,每个重复36只蛋鸡。对照组蛋鸡饲喂基础日粮,试验组分别在基础日粮中添加0.5%、1.0%、1.5%杜仲叶粉。预试期1 w,正式试验期7 w。结果显示,1.0%组、1.5%组蛋鸡的产蛋率均显著高于对照组(P<0.05)。1.0%组鸡蛋的蛋黄颜色显著高于对照组(P<0.05)。1.0%组、1.5%组蛋鸡血清总胆固醇(TC)含量均显著低于对照组(P<0.05)。各试验组蛋鸡血清总抗氧化能力、过氧化氢酶活性显著高于对照组(P<0.05)。1.0%组、1.5%组蛋黄胆固醇含量均显著低于对照组(P<0.05)。研究表明,日粮中添加杜仲叶粉可以改善蛋鸡的产蛋率和蛋品质,降低血清总胆固醇和蛋黄胆固醇含量,提高抗氧化能力,杜仲叶粉推荐添加量为1.0%。

杜仲叶提取物绿色合成纳米铂颗粒及其美白作用

以杜仲叶提取物为还原剂和稳定剂,采用一步法绿色合成纳米铂颗粒(PtNPs),通过单因素试验分析反应条件对PtNPs颜色和浓度的影响,并研究PtNPs对酪氨酸酶活性的影响。结果表明:在一定范围内改变反应条件可调控合成PtNPs的颜色和浓度,适宜反应条件为反应时间60 min、反应温度90℃、氯铂酸浓度3.5 mmol/L、杜仲叶提取物质量浓度10.0 mg/mL,在该条件下制备的PtNPs粒径平均尺寸为(2.27±0.65) nm,具有面心立方结构,分布较均匀;PtNPs对体外蘑菇酪氨酸酶活性有较强的抑制作用,IC50为(6.98±0.76)μg/mL,显著降低了A375细胞内酪氨酸酶活性和UVB诱导的黑色素含量,可达到美白皮肤的效果。

基于肾主骨理论探讨杜仲通过调控OPG/RANKL/RANK通路对去势骨质疏松大鼠的影响

目的基于肾主骨理论探讨杜仲对去势骨质疏松大鼠的影响。方法大鼠分为对照组、模型组、阿法骨化醇组(0.05μg/kg)、杜仲组(2.76 g/kg),每组20只,除对照组外,其余组大鼠建立去势骨质疏松性骨折模型,灌胃给予相应药物,12周后检测各组大鼠骨密度、血清骨生成指标、尿液骨代谢指标,以及OPG、RANKL、RANK mRNA和蛋白表达。结果与对照组比较,模型组大鼠骨密度、股骨质量和压碎力、血清ALP活性、骨钙素水平以及骨组织OPG、RANKL、RANK mRNA和蛋白表达均降低(P<0.05),尿液钙和磷水平升高(P<0.05);与模型组比较,阿法骨化醇组和杜仲组大鼠骨密度、股骨质量和压碎力、血清ALP活性、骨钙素水平以及骨组织OPG、RANKL、RANK mRNA和蛋白表达均升高(P<0.05),尿液钙和磷水平降低(P<0.05);与阿法骨化醇组比较,杜仲组OPG、RANKL、RANK mRNA和蛋白表达升高(P<0.05),尿液钙和磷水平降低(P<0.05)。结论杜仲可通过减少钙磷流失,激活OPG/RANK/RANKL通路,进而缓解大鼠骨质疏松症状,起到补肾生骨作用。

杜仲水提物改善动物睡眠障碍作用机制的网络药理学分析及实验动物验证

利用网络药理学分析方法,经过动物试验验证,探究杜仲改善小鼠睡眠障碍的作用机制。通过TCMSP数据库获得杜仲的潜在作用靶点,利用OMIM和GeneCards数据库获得睡眠障碍潜在靶点。将两个靶点合集交集后构建蛋白互作网络,筛选核心靶点。通过David在线平台进行基因本体功能(GO)富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。使用Cytoscape软件构建杜仲成分-睡眠障碍靶点-通路网络,之后采用分子对接技术进行检验。最终通过PCPA小鼠睡眠障碍模型进行体内试验验证。网络药理学和分子对接结果显示,杜仲有效成分烟碱单宁、表奎宁定等通过调节多巴胺D2受体基因(DRD2)和GABAA受体α亚基基因(GABRA1)的表达,发挥镇静安神、抗焦虑的作用。动物试验结果显示,杜仲显著减少了睡眠障碍小鼠的运动量(P<0.05),下调睡眠障碍小鼠下丘脑组织中DRD2基因的表达(P<0.01),上调GABRA1基因的表达(P<0.01)。预测并验证了杜仲改善睡眠障碍的作用机制,为其临床应用提供了一定的参考依据。

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。

杜仲水提物诱导大鼠肾上腺嗜铬细胞瘤细胞向神经元样细胞分化

目的 探讨杜仲水提物对大鼠肾上腺嗜铬细胞瘤细胞(Rat pheochromocytoma cells, PC12细胞)向神经元样细胞分化的作用。方法 用0~10 mg·mL^(-1)杜仲水提物培养PC12细胞4 d,双抗噻唑蓝法(MTT法)检测吸光度(A值)并选取最佳浓度配置杜仲诱导培养基;诱导4 d后通过免疫荧光标记神经丝蛋白H(neurofilament-H,NF-H)检测诱导情况;通过免疫蛋白印迹检测细胞外信号调节激酶(extracellular signal-regulated kinase, ERK)及磷酸化细胞外信号调节激酶(phosphorylated extracellular signal regulated kinases, p-ERK)表达情况。结果 不同浓度杜仲培养4 d的PC12细胞,与空白对照A值相比,1~3 mg·mL^(-1)剂量组有增高趋势;4~7 mg·mL^(-1)剂量组有降低趋势;8~10 mg·mL^(-1)剂量组显著降低(P<0.05)。4 mg·mL^(-1)含杜仲诱导培养基培养PC12细胞4 d, 90%以上分化为神经元样细胞,细胞胞体变大,突起伸长,互相连接成网,约20%突起长度> 30μm,显著高于对照组。随着诱导时间的延长,NF-H表达逐渐增多增强,诱导4 d后,约90%以上的细胞NF-H强阳性表达,p-ERK蛋白表达水平较对照组明显降低。结论 杜仲水提物能够诱导PC12细胞分化为神经元样细胞,表达神经元特异蛋白神经丝蛋白,其机制可能与ERK信号通路调节密切相关。

Pharmacokinetic study about compatibility of Eucommia ulmoides and Psoralea corylifolia

Objective:The compatibility of Eucommia ulmoides(Eu)and Psoralea corylifolia(Pc)on the pharmacokinetic(PK)properties in the rat was explored in this study.Methods:Eu extract,Pc extract and the combined extracts(crude drug ratio was 2:1)was administered by gavage,respectively.Two PK experiments were conducted.In first one,the blood samples were collected via the occuli chorioideae vein to get the PK properties of the components.In second one,the blood samples were simultaneously collected via the internal jugular vein or portal vein at different time points and the concentrations of target ingredients were detected by LC/MS/MS to clear the location where the interaction of Eu and Pc took place in vivo.Results:Eight of 11 ingredients in Eu and Pc extract were determined in rat plasma.The exposure levels of geniposidic acid(GPA),aucubin(AU),geniposide(GP),pinoresinol diglucoside(PDG),psoralen glycosides(PLG)and isopsoralen glycosides(IPLG)were decreased 1/5–2/3 after administration of combined extracts.Comparing to the combined administration,the exposure of GPA and AU in plasma of single Eu administration collected via the portal vein were decreased 1/3–2/3,and the values of AUC0-24h and AUC0-∞ of GP collected from the portal vein or internal jugular vein were double increased.The other components’parameters were not significantly changed.Conclusion:In summary,the Pc and Eu combined administration could affect the exposure of the main components of Eu extract in rats due to the changed intestinal absorption.The research on the compatibility of Pc and Eu was helpful to guide the clinical administration of Eu and Pc simultaneously.