Dietary Epimedium extract supplementation improves intestinal functions and alters gut microbiota in broilers

Background Growth-promoting antibiotics have been banned by law in the livestock and poultry breeding industry in many countries.Various alternatives to antibiotics have been investigated for using in livestock.Epimedium(EM)is an herb rich in flavonoids that has many beneficial effects on animals.Therefore,this study was planned to explore the potential of EM as a new alternative antibiotic product in animal feed.Methods A total of 7201-day-old male broilers(Arbor Acres Plus)were randomly divided into six groups and fed basal diet(normal control;NC),basal diet supplemented with antibiotic(75 mg/kg chlortetracycline;CTC),and basal diet supplemented with 100,200,400 or 800 mg/kg EM extract for 6 weeks(EM100,EM200,EM400 and EM800 groups).The growth performance at weeks 3 and 6 was measured.Serum,intestinal tissue and feces were collected to assay for antioxidant indexes,intestinal permeability,lactic acid and short-chain fatty acids(SCFAs)profiles,microbial composition,and expression of intestinal barrier genes.Results The average daily feed intake in CTC group at 1-21 d was significantly higher than that in the NC group,and had no statistical difference with EM groups.Compared with NC group,average daily gain in CTC and EM200 groups increased significantly at 1-21 and 1-42 d.Compared with NC group,EM200 and EM400 groups had significantly decreased levels of lipopolysaccharide and D-lactic acid in serum throughout the study.The concentrations of lactic acid,acetic acid,propionic acid,butyric acid and SCFAs in feces of birds fed 200 mg/kg EM diet were significantly higher than those fed chlortetracycline.The dietary supplementation of chlortetracycline and 200 mg/kg EM significantly increased ileal expression of SOD1,Claudin-1 and ZO-1 genes.Dietary supplemented with 200 mg/kg EM increased the relative abundances of g_NK4A214_group and Lactobacillus in the jejunal,while the relative abundances of Microbacterium,Kitasatospora,Bacteroides in the jejunal and Gallibacterium in the ileum decreased.Conclusion Supplementation with 200 mg/kg EM extract improved the composition of intestinal microbiota by regulating the core bacterial genus Lactobacillus,and increased the concentration of beneficial metabolites lactic acid and SCFAs in the flora,thereby improving the antioxidant capacity and intestinal permeability,enhancing the function of tight junction proteins.These beneficial effects improved the growth performance of broilers.

朝鲜淫羊藿(Epimedium koreanum Nakai)不同生境种群生物量与更新潜力

朝鲜淫羊藿(Epimedium koreanum Nakai)为重要药用植物资源,由于种子繁殖问题较大,而尚未实现真正的人工栽培,野生资源在长期过度利用中已遭到一定的破坏。研究结果表明:朝鲜淫羊藿的有性生殖能力较弱,平均开花和结果率分别不超过8%和6%,最低开花和结果率均在次生杂木林下,分别仅有2%和0.8%。朝鲜淫羊藿主要以无性繁殖维持种群更新和增长,其中,2龄根茎无性繁殖活力最高,其产生的潜伏芽和1龄根茎数量平均占80%以上;3龄和4龄根茎产生的潜伏芽和1龄根茎数量显著减少,而4龄根茎的潜伏芽100%不能发育成1龄根茎;5龄以后的根茎不再有无性繁殖活力。朝鲜淫羊藿无性繁殖潜伏芽数量具有较大冗余性,并随着根茎龄级的增高和生境条件的变差其冗余性也增大。另外,朝鲜淫羊藿的开花率、结果率和根茎产生潜伏芽及1龄根茎数量,均随生境条件的不适宜性增强和根茎龄级增大而变异系数明显增大,说明生境条件变差和生理功能衰退导致其种群增殖的不确定性增加,而有序性降低。总之,朝鲜淫羊藿是极易遭到利用威胁的易危物种,应注意对野生资源利用强度和方式的研究与控制。

箭叶淫羊藿(Epimedium sagittatum Maxim)种子后熟作用的形态解剖研究

目的:了解箭叶淫羊藿种子后熟休眠的形态结构基础,探索其种子的萌发条件。方法:在对箭叶淫羊藿种子进行常规发芽试验的基础上,采用直接测量和石蜡制片技术等方法对其外部形态和解剖结构进行了观察。结果:箭叶淫羊藿种子存在明显的休眠现象,其外部形态无异常,但其胚结构未分化,停留在球型胚阶段。后熟过程中,其胚结构逐步发育完全,形成子叶胚。结论:箭叶淫羊藿种子存在明显的休眠现象,其休眠类型为形态生理休眠(MPD)。其萌发需要一定时间的层积以完成其胚分化。

川产淫羊藿属(Epimedium)药用植物的无性系构件特征和生物量配置比较

在构件水平上,对川产的3种药用淫羊藿属植物(淫羊藿(Epimedium brevicornu)、箭叶淫羊藿(E.sagitta-tum)、柔毛淫羊藿(E.pubescens))的无性系构件特征、克隆构型及构件生物量配置进行比较,结果表明在野生状态下:该属3个种无性系构件的形态特征差异显著,且存在较大的变异性,变异系数范围在29.29%～48.31%之间;箭叶淫羊藿更趋于"游击型"克隆构型,而淫羊藿和柔毛淫羊藿则更趋于"密集型"克隆构型;该属3个种都将高比例生物量配置到叶片或者根茎,其次为茎,再其次为根。柔毛淫羊藿具有更强的克隆繁殖能力,单位面积上各构件生物量均最大,应作为人工引种栽培的首选种。

Network Pharmacology Analysis of Epimedium brevicornu(Yinyanghuo)for Treatment of Ovarian Cancer

[Objectives]To elucidate potential targets and mechanisms of action of Epimedium brevicornu in treating ovarian cancer.[Methods]The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform was used to screen active components of E.brevicornu for disease control and prevention,and potential targets were collected from the DisGeNET database.These sets of bioactive and targets were analyzed using Ingenuity Pathway Analysis(IPA)to predict molecular networks affected by E.brevicornu in ovarian cancer.Venny 2.1.0 software was used to screen for proteins affected by interactions between disease and active components,which were input into the STRING 11.0 platform to construct a protein-protein interaction network.Then IPA and STRING were used to analyze common targets which were obtained from the two data analysis platform.[Results]A total of 23 major active components of E.brevicornu and 200 potential human targets were screened.IPA analysis identified 363 pathways and 24 networks shared between the set of predicted Yinyanghuo targets and ovarian cancer-associated proteins.These pathways are involved mainly in molecular mechanisms of cancer,glucocorticoid receptor signaling pathways,pancreatic adenocarcinoma signaling pathways,aryl hydrocarbon receptor signaling pathways,and macrophage function.The 24 networks have been implicated mainly in cancer,endocrine system disorders,body damage and abnormality,cell growth and proliferation,connective tissue development and function,tissue development,and other biological functions.IPA and STRING combined analysis suggested that AKT1,CASP3,JUN,FOS and CCND1 are the most likely targets of Yinyanghuo in treating ovarian cancer.[Conclusions]Our network pharmacology analysis identified several pathways that Yinyanghuo may influence to reduce ovarian cancer risk;in particular,it identified specific protein targets,including AKT1,CASP3,JUN,FOS and CCND1.

EFFECT OF EPIMEDIUM FLAVONE TOTAL(EFT) ON THE IMMUNOFUNCTION OF EHRLICH ASCITES CANCER(EAC) BEARING MICE

Effect of Epimedium Flavone Total(EFT) on the immunofunction of mice which was burdened with Ehrlich Ascites Cancer (EAC) was studied. The results showed that EFT could increase phagocytosis activity of macrophagocyte (P<0 05), promote lymphocyte transformation rate (P<0 01), increased the sero antibody level meanly (P<0 05), low dosage group could increased Plaque Forming Cell (PFC) signficantly (P<0 05), raise the ratio of spleen to body (P<0 05) of EAC bearing mice. EFT may be enhance burdened mice’s immunofunction. Low dosage can improve sero antibody level. Effect on regulating cellular immunofunction are related to dosage. EFT may be inhibit the growth of EAC cells to some degree.

Mechanism of "Epimedium-Cistanche deserticola" in the treatment of breast cancer with bone metastasis

Objective:"Epimedium - Cistanche deserticola" is a kind of kidney tonifying drug commonly used in the treatment of breast cancer bone metastasis, which has good clinical effect, but the pharmacological mechanism has not been fully clarified. Methods: In this study, the network pharmacology and bioinformatics technology were used to explore the mechanism of "Epimedium - Cistanche deserticola" in the treatment of breast cancer bone metastasis. TCMSP, TCM database@Taiwan and TCMID databases were used to screen the main effective components of the drug. Swiss Target Prediction and STITCH databases were used to search the potential target of action of Epimedium and Cistanche deserticola. Genecards, OMIN and Drugbank databases were used to search the cause of bone metastasis of breast cancer. The target of action of the drug and the disease gene were mapped for GO and KEGG signal pathway analysis, A visualized network of "drug - component - target - signaling pathway" was constructed by using the software of Cytoscape 3.6.0, and the core genes were screened out. Results: The study found that there are 30 main effective components of Epimedium and Cistanche deserticola, and 544 genes are involved in the potential therapeutic targets, among which 101 genes are potential targets of Epimedium and Cistanche deserticola in the treatment of breast cancer bone metastasis. Through the analysis of GO and KEGG pathways, we found that the mechanisms involved in antitumor, osteoblast differentiation, osteoclast apoptosis and regulation of bone microenvironment, such as apoptosis, osteoclast differentiation, PI3K-Akt, HIF-1 signaling pathway, T cell receptor signaling pathway, etc. TP53, VEGFA, AKT1, EGFR, SRC, CCND1, MAPK3, ESR1 may be the key genes in the treatment of breast cancer bone metastasis. Conclusion: In this study, the network of "drug - component - target- signaling pathway" was constructed through network pharmacology, and it was found that the mechanism of "Epimedium - Cistanche deserticola" in the treatment of breast cancer bone metastasis involves multiple targets and pathways, which is conducive to guiding clinical medication.

Isolation and Extraction of Total Flavonoids from Epimedium Koreanum Nakai by Supercritical Fluid Extraction

In the present paper is reported the method for the isolation and extraction of total flavonoids of Epimedium Koreanum Nakai by means of supercritical fluid extraction(SFE). By examining pressure, temperature, amounts of modifier and extraction time, the optimized condition of SFE is confirmed as 30 MPa and 60 ℃, with 70% ethanol as the modifier. The samples were statically extracted for 30 min, followed by dynamic extraction for 120 min at a flow rate of 6 mL/min. The quantitative analysis of total flavonoids was performed by UV-Vis spectrophotometry. Compared with the conventional method, the SFE method is more efficient, more rapid and more friendly environmentally.

Simultaneous determination of seven flavonoids in Epimedium by liquid chromatography-tandem mass spectrometry method

In this paper,a sensitive and specific liquid chromatography-electrospray ionisation-mass spectrometry (LC-ESI-MS) method has been developed and validated for the identification and determination of seven flavonoids,namely epimedin A,epimedin B, epimedin C,icariin,sagittatoside B,2″-O-rhamnosyl icarisideⅡ,and baohuosideⅠin Epimedium from different sources.

Studies on the Effects of Epimedium Extract on Erythrocytes

The effect of Epimedium extract (EE) on erythrocytes was investigated by atomic force microscopy (AFM). The images of the surface structures showed clear concave and progressive increase of surface roughness of erythrocyte after incubation with EE at concentration of 0.2 or 0.05 g/L, far below its critical hemolytic levels. The AFM results also indicated that the granules of the fine surface structure increased, which caused by aggregation of membrane protein. Further study showed that the change in surface topography of erythrocyte membrane might be connected with the increase of intracellular free Ca2+ induced by EE.

A New Flavone Glycoside from Epimedium koreanum

A novel flavone glycoside named 3’’’-carbonyl-2’’-β-L-quinovosyl icariside II was isolated from the aerial parts of Epimedium koreanum. Its chemical structure was elucidated on the basis of spectral analysis.

Qualitative and Quantitative Evaluation of Epimedium and Ginseng Contained Combinations Using HPLC

Aim:To develop a rapid,effective method for the detemination of flavonoids and ginsenosides in one injection and evaluate the flavonoids and ginsenosides content to control the ratio of Epimedium and Ginseng herbs in botanical combinations.Methods: The quality evaluation was determinatted using reversed-phase high performance liquid chromatog raphy(HPLC),referred by the major flovonoids from Epimedium,epimedin A,epimedin B,epimedin C,and icariin as the standards,and the major ginsenosides Rg1,Re,Rf,Rb1,Rb2,and Rd as the standards,included Epimedium brevicornum Maxim.,E.sagittatum(Sieb.et Zucc)Maxim.,E.koreanum NaKai,P.ginseng C.A.Meyer,P.quinquefolium L.,P.notoginseng and some products containing the above herbs.Results:The main flavonoids and ginsenosides could be clearly resolved in the single analysis.Conclusion.The results can be effectively used in evaluating qualitatively and quantitatively the ration of Epimdium and Ginseng contained products.

淫羊藿属(Epimedium)植物系统分类研究进展

Epimedium因其独特的地理分布、杂交可育、种内变异等特点,其种间界定仍模糊不清,被认为是分类和系统发育上最具挑战和困难类群之一。通过对相关文献资料及标本数据库的查阅与整理,从分类方法对该属淫羊藿属药用植物进行了总结分析,并从分子生物学、生物信息学、谱系地理学等现代技术对该属的分类鉴定、种间关系、起源与演化方面进行了展望,从而为Epimedium进一步研究及开发利用提供参考。

Two new prenylflavonoids from Epimedium sutchuenense

二新 prenylflavonoids，命名 sutchuenmedin A (1 ) 和 sutchuenmedin B (2 ) ，从 Epimedium sutchuenense 的 70% EtOH 摘录被孤立。他们的结构被决定从光谱分析。

Exploring the effect and mechanisms of Epimedium on diabetic testicular injury using network pharmacology,molecular docking,and cell experiments

Background:Studying the potential targets and mechanisms of Epimedium for anti-diabetic testicular injury using network pharmacology,molecular docking,and cell experiments.Methods:Acquisition of major components and targets of Epimedium was based on TCMSP,TCMID,and Symmap databases and retrieval of diabetic testicular injury targets by OMIM,GeneCards,Pharmgkb,and Drugbank databases.Intersecting targets were obtained from the Venny 2.1.0 database and input SRTING data to construct a protein-protein interaction(PPI)network,and key targets were screened in Cytoscape 3.8.0 software.Then the Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses of intersecting targets were conducted through the DAVID database.Further,AutoDock software was used to verify docking between the main components and the core target proteins.In addition,a Cell Counting Kit-8(CCK-8)assay was used to determine the survival effect of quercetin,the main component of Epimedium,on TM4 sertoli-like cells exposed to palmitic acid(PA).Results:Quercetin,kaempferol and luteolin in epimedium were identified as the main components in the treatment of diabetic testicular injury.It has core target proteins including MMP9,AKT1,and TNF.The biological process mainly involves the regulation of the apoptotic signaling pathway.The key pathways of KEGG are the AGE-RAGE signaling pathway in diabetic complications,PI3K-Akt and MAPK signaling pathway.Molecular docking results showed that quercetin had the strongest binding ability to MMP9.Also,PA-challenged cells had a lower survival rate,which was alleviated by the administration of quercetin.Conclusion:Our findings suggest that Epimedium attenuates diabetes mellitus(DM)-induced testicular injury through AGE-RAGE,PI3K-Akt and MAPK signaling pathway.These insights offer a potential therapeutic strategy for managing DM-induced testicular injury,will be the basis for future clinical research.

基于不同炮制方法的淫羊藿黄酮类成分含量研究

试验旨在寻求羊脂油炮制淫羊藿的内在机理,以甘肃产淫羊藿为研究对象,用羊脂油、牛脂油、猪脂油进行炮制,分别提取有效成分,利用紫外分光光度法和高效液相色谱法进行有效成分含量测定,比较不同炮制品和生品总黄酮、淫羊藿苷、宝藿苷Ⅰ和朝藿定C的含量。结果显示:各炮制品总黄酮含量均显著低于生品(6.84%),其中羊脂油炙含量(5.69%)显著高于其他炮制品;生品和羊、牛脂油炙炮制品中淫羊藿苷含量无显著差异,羊脂油炙炮制品为0.77%,牛脂油炙炮制品为0.70%,生品为0.70%,猪脂油炙炮制品中淫羊藿苷含量为0.50%,较生品和其他炮制品显著降低;羊脂油炙、牛脂油炙淫羊藿中宝藿苷Ⅰ含量无显著性差异,分别为0.08%和0.07%,但显著高于生品(0.05%)和猪脂油炙淫羊藿(0.04%);羊、牛、猪脂油炙炮制品中朝藿定C含量分别为0.29%、0.27%和0.24%,均显著低于生品(0.57%)。得出结论:淫羊藿最佳炮制辅料为羊脂油,其次为牛脂油。

淫羊藿治疗骨质疏松骨折的网络药理学分析

目的利用网络药理学原理探讨淫羊藿治疗骨质疏松骨折的分子作用机制。方法利用中药系统药理学分析平台(TCMSP数据库)筛选淫羊藿的主要活性成分和作用靶点,应用GeneCards数据库、OMIM数据库、PharmGkb数据库、TTD数据库、DrugBank数据库预测骨质疏松骨折的相关靶点,制作维恩图获取淫羊藿与骨质疏松骨折的共同靶点,运用Cytoscape软件绘制中药-成分-疾病-靶点调控网络,利用STRING数据库进行蛋白相互作用网络的构建与分析。最后,使用R语言和Bioconductor平台进行基因本体富集分析(GO)和基因相互作用通路分析(KEGG)。结果淫羊藿主要是通过23种活性成分、121个作用靶点及155个生物学通路发挥作用治疗骨质疏松骨折。其中主要活性成分为槲皮素、山奈酚和木犀草素;主要核心靶点有AKT1、TP53、Jun、HSP90AA1和CASP3等;关键通路为脂质和动脉粥样硬化信号通路。结论通过网络药理学揭示了淫羊藿通过多成分、多靶点、多通路治疗骨质疏松骨折的作用特点,为进一步的研究和实验工作提供了基础。

淫羊藿总黄酮对去势大鼠抗酒石酸酸性磷酸酶5b的影响及与骨密度的相关性

目的:探讨淫羊藿总黄酮(TFE)对去势大鼠抗酒石酸酸性磷酸酶5b(TRACP-5b)及骨密度的影响。方法:10月龄雌性SD大鼠去卵巢法建立大鼠模型,随机数字表法分为空白对照组(N)、模型对照组(O)、TFE低(T_(L))、中(T_(M))、高(T_(H))剂量组,β-雌二醇(E)组各10只,灌胃干预90d。用抗酒石酸酸性磷酸酶试剂盒测定TRACP-5b含量,用小动物分析软件测定活体全身骨密度。结果:(1)TRACP-5b水平O组明显高于N组,具有显著性差异(P<0.01),T_(L)、T_(M)、T_(H)和E组TRACP-5b水平明显低于O组,具有显著性差异(P<0.01)。(2)与N组比较,O、T_(L)、T_(M)、T_(H)和E组腰椎骨密度均下降,唯有O组呈显著性差异(P<0.05);与O组比较,T_(M)、T_(H)组腰椎骨密度升高、呈显著性差异(P<0.05),E组腰椎骨密度升高、具有显著性差异(P<0.01)。(3)与N组比较,O、T_(L)、T_(M)、T_(H)和E组全身骨密度均下降,唯有O组差异有统计学意义(P<0.05);与O组比较,T_(L)、T_(M)、T_(H)和E组全身骨密度均升高,唯有E组呈显著性差异(P<0.05)。结论:TFE可以降低去势大鼠血清中TRACP-5b的含量,并与去势大鼠腰椎骨密度、全身骨密度呈一定的量效关系。

淫羊藿中异戊烯基黄酮的鉴别和含量测定的综合实验设计

为加深学生对传统中药淫羊藿的药效成分异戊烯基黄酮的认识,设计了综合化学实验.将淫羊藿粉末经乙醇热回流提取,蒸馏浓缩得到总浸膏,再将总浸膏溶于水,依次用石油醚、乙酸乙酯和正丁醇萃取,得到石油醚相、乙酸乙酯相和正丁醇相.将三相的有机物分别用薄层色谱展开,在紫外灯下观察254 nm和365 nm荧光颜色和强度,同时用硫酸、氯化铁显色,表明各相均含有黄酮.以淫羊藿苷为标品,通过作标准曲线结合紫外光谱对其各相的异戊烯基黄酮含量进行平行测定,同时考察了方法的重复性和样品的稳定性.

补骨脂-淫羊藿对类风湿关节炎抗炎机制的分子对接分析:动物实验验证

背景:临床上补骨脂-淫羊藿治疗类风湿关节炎疗效明显,但两者所含有效成分复杂,在分子水平上治疗类风湿关节炎的作用机制仍不明确。目的:基于网络药理学和分子对接技术建立胶原诱导型关节炎模型,验证补骨脂-淫羊藿治疗类风湿关节炎可能的作用靶点及通路,为以补骨脂-淫羊藿为主的临床方剂使用提供可靠的实验依据。方法:借助中医药研究平台、中医百科全书和上海有机所的中药与化学成分数据库等检索并筛选有效成分,从PubChem平台获取3D分子式,通过PharmMapper和SwissTargetPrediction平台进行靶标预测;结合DrugBank、GeneCards、OMIM等基因数据库完成类风湿关节炎的疾病靶点获取,经Uniport数据库校准靶标,借助VENNY 2.1获取补骨脂-淫羊藿与疾病交集靶点并绘制韦恩图;采用STRING平台构建蛋白质互作网络图;使用Metascape平台进行基因本体论功能分析及京都基因与基因组百科全书分析,进行数据可视化,利用Cytoscape 3.9.0构建中药-成分-靶点-疾病-通路四重网络模型;运用AutoDock-Vina软件将主要有效成分与核心靶点进行分子对接验证,探索最佳结合靶点。建立Ⅱ型胶原+佐剂诱导型关节大鼠模型,用补骨脂-淫羊藿干预21 d后,观察其对相关通路靶点及炎性细胞因子的影响。结果与结论:①筛选补骨脂与淫羊藿活性成分28个,与类风湿关节炎交集靶点共288个,主要成分有异补骨脂素、补骨脂定、淫羊藿苷等;交集靶点主要有丝氨酸/苏氨酸蛋白激酶1(AKT1)、肿瘤坏死因子、血管内皮生长因子A等;②基因本体论分析获得生物过程2232条,主要与丝氨酸蛋白磷酸化、AKT正调控、活性氧代谢过程等功能有关;③京都基因与基因组百科全书富集分析结果202条,主要有PI3K/AKT信号通路和表皮生长因子受体信号通路等,可能通过调节滑膜细胞凋亡与增殖、抑制炎性因子等发挥治疗作用;④分子对接结果表明补骨脂-淫羊藿主要与AKT1及雌激素受体转录因子1结合活性最强,并形成稳定结构,与PI3K/AKT等凋亡增殖、炎性介导等调控信号通路密切相关;⑤补骨脂-淫羊藿可降低胶原诱导型关节炎大鼠模型血清中白细胞介素1β、白细胞介素6、肿瘤坏死因子α的表达;⑥补骨脂-淫羊藿可调低胶原诱导型关节炎大鼠模型关节滑膜中p-PI3K、p-AKT、p-FOXO1蛋白的表达;⑦结果证明,补骨脂-淫羊藿可能经PI3K/AKT/FOXO1信号通路抑制关节滑膜细胞增殖和抑制炎性因子表达等发挥治疗作用,这可能与类风湿关节炎关节炎症和骨破坏的发生密切相关,同时为临床的合理使用及新药开发提供了参考依据。