Wedelolactone from Eclipta prostrata(L)L.suppresses inflammation and improves insulin resistance

Objective:To evaluate the effects of wedelolactone,a major flavonoid from Vietnamese Eclipta prostrata(L)L.,on inflammation and insulin resistance.Methods:Wedelolactone was extracted from the leaves of Vietnamese Eclipta prostrata(L.)L.with methanol by Soxhlet.The effects of wedelolactone on lipopolysaccharide(LPS)-induced cytokine production,reactive oxygen species(ROS)generation,and nicotinamide adenine dinucleotide phosphate(NADPH)oxidase activities in Raw 264.7 cells were measured by enzyme-linked immunosorbent assay(ELISA),specific immunofluorescent dyes and luminometric analysis,respectively.In addition,its effects on glucose uptake and the protein expression of insulin receptor substrate 1(IRS1)and glucose transporter 4(GLUT4)were examined in 3T3-L1 cells by immunofluorescent dyes and Western blot.Results:Wedelolactone at 30μg/mL significantly inhibited LPS-induced production of tumor necrosis factor-α,interleukin(IL)-6,and IL-8(P<0.01)with no noticeable effects on IL-10 level.It also reduced ROS generation and NADPH oxidase activities in LPS-stimulated Raw 264.7 cells(P<0.01).Furthermore,wedelolactone showed anti-insulin resistance activity,as evidenced by improved glucose uptake and the upregulated expression of IRS1 and GLUT4 in 3T3-L1 cells(P<0.01).Conclusions:Wedelolactone exhibits anti-inflammation and anti-insulin resistance effects,which may be used for the treatment of diabetes and inflammation-associated diseases.

Characterization of Endophytes Isolated from <i>Eclipta prostrate</i>and Roles in Regulating the Gut Microbiota of C57BL/6J Mice

Eclipta prostrata has been extensively studied for its promoting effect on human health. Previous study proved that E. prostrata possessed anti-osteoporosis function in SAMP6 mice via gut microbiota (GM). Endophytes are widespread in plants, especially in Chinese herbal medicines. However, little is known regarding the endophytes of E. prostrata. In this study, we focus on screening and identifying the endophytes from plant E. prostrata and exploring their roles in modulating GM. According to biochemical, physiological tests and 16S rDNA sequence analysis, ten endophytes were characterized in different locations of plant E. prostrata belonging to Lactococcus, Bacillus, Enterococcus, Exiguobacterium and Pantoea. The antimicrobial activity of endophytes EP01-10 was investigated via the oxford cup method. Furthermore, the acid and bile salt resistant ability of EP01-10 was detected to explore their survival ability in gastrointestinal tract. Results indicated that strains of Lactococcus, Bacillus and Exiguobacterium (EP01, 03 and 05) showed strong antimicrobial activity against Escherichia coli and Staphylococcus aureus and strains of Lactococcus, Bacillus and Enterococcus (EP01, 02, 04, 08 and 10) have the ability to survive in the gastrointestinal tract. 16S rRNA sequencing of GM in C57BL/6J mice was performed for diversity and structure analysis responded to the administration of E. prostrata extract. E. prostrata extract acted on improving the microflora diversity, particularly in increasing the abundance of Lactococcus significantly. Thus, as an endophyte, Lactococcus plays an important role in E. prostrata modulating intestinal tract. Our study suggested that endophytes of Chinese herbal medicines might be a novel target for the treatment of diseases by regulating the GM.

HPLC法测定不同产地墨旱莲药材中木犀草素含量

目的建立墨旱莲药材中木犀草素的高效液相（HPLC）含量测定方法。方法色谱柱：Phenomenex Luna C18柱（4.6 mm×250 mm,5μm）,流动相：乙腈-0.1%磷酸（25∶75）,流速：1.0 mL/min,检测波长：348 nm,柱温：30℃。结果木犀草素在57~1 140 ng（r=1.000）范围内呈良好的线性关系,平均加样回收率为96.64%,RSD为1.87%。结论该方法简便、准确,可以作为墨旱莲中木犀草素含量测定的一种有效方法。

UHPLC-Q-TOF/MS技术应用于中药旱莲草化学成分研究

目的通过超高效液相色谱-四极杆飞行时间串联质谱(UHPLC-Q-TOF/MS)对中药旱莲草的化学成分进行系统、全面研究。方法采用Agilent SB-C18(2.1 mm×100 mm,1.8μm)柱,流动相为乙腈-甲酸溶液系统梯度洗脱,质谱采用ESI正离子模式,分别对桂林阳朔、桂林荔蒲、湖南邵阳、南宁和湖北襄樊五个产地的旱莲草药材进行分析。结果该方法共鉴别出旱莲草中29个化学成分。结论本实验为建立旱莲草药材的质量控制标准提供了参考依据。

UHPLC-Q-TOF/MS法测定中药旱莲草中五种指标性化学成分的含量

目的建立不同产地旱莲草药材中的5个指标性成分含量测定方法。方法采用UHPLC—Q—TOF／MS技术，对桂林阳朔，广西荔蒲，湖南邵阳，广西南宁和湖北襄樊等5个产地旱莲草药材中的5个指标性成分进行含量测定与分析。结果旱莲草药材中芹菜素在2．012～402．4μg·ml～、十六烷酸在2．508~501．6lug·ml-1、木樨草素在1．252~250．4μg·ml-1、蟛蜞菊内酯在1．664～332．8μg·ml-1、木樨草苷在1．080～216．0μg·ml-1范围内，各待测成分的提取离子色谱峰面积和对应的浓度均具有良好的线性关系，方法的精密度及稳定性良好。结论不同产地的旱莲草药材在指标性成分的含量上存在一定差异。

室内模拟旱直播稻田环境下齐整小核菌Sclerotium rolfsii菌株SC64致病力的影响因子及除草效果的研究

本文通过温室盆栽试验,以鳢肠Eclipta prostrata为例,研究杂草叶龄、不同含菌丝体基质接种量以及温度、土壤湿度、保湿期时间等环境因子对菌株SC64侵染鳢肠的影响。在此基础上,温室条件下模拟旱直播水稻田环境下菌株SC64防治节节菜Rotala indica、异型莎草Cyperus difformis、鸭舌草Monochoria vaginalis和耳叶水苋Ammannia arenaria的试验。结果表明,鳢肠幼苗在5叶期以下,接种含菌丝体固体基质剂量在100g m 2以上,温度范围27～33℃,土壤相对饱水90%左右,并且接菌后湿度保持至少24 h,是菌株SC64达到理想除草效果的必需条件。温室模拟试验结果显示,接种含菌丝体固体基质剂量在120 g m 2时,28 d可引起节节菜和鸭舌草81%和74%的植株死亡率及83.8%和82.5%的鲜重抑制率;对异型莎草和耳叶水苋的防效稍差,仅能引起约50%的植株死亡率及60%～65%的鲜重抑制率。这表明菌株SC64可有效控制阔叶杂草,具有开发作为生物除草剂的潜力。

HPLC法测定墨旱莲中两个噻吩类化合物的含量

目的运用HPLC法测定12个不同产地墨旱莲药材中α-三联噻吩甲醛和α-三联噻吩甲醇的含量。方法色谱柱为Agilent Zorbax SB-C18柱(250 mm×4.6 mm,5μm);流动相为乙腈和体积分数为0.5%的甲酸溶液,采用梯度洗脱程序;流速为1 m L·min-1;检测波长为395 nm;柱温为25℃;进样量为20μL。结果两个噻吩类化合物在12 min内达到基线分离,线性良好;方法学考察过程中,日内和日间精密度、最低定量限和最低检测限均符合相关标准,加样回收率(n=3)分别为101.17%和100.95%;用该方法测定了12个不同产地墨旱莲药材中两个噻吩类化合物的含量。结论该方法可用于对墨旱莲药材进行质量控制。

Biological and Functional Properties of Wedelolactone in Human Chronic Diseases

Medicinal herbs are well known and studied over the past millennia in most of the developing countries as a rational means of treatment against various diseases and disorders.Wedelolactone(WDL),a major bioactive compound in Eclipta prostrata L(Eclipta alba L),has been reported with potential benefits in human health against chronic diseases.However,a comprehensive study on WDL pharmacological benefits in various ailments,to the best of our knowledge,is not yet reported.Thereof,the present review provides the recent therapeutic applications in reference to biological and functional activities against major human chronic diseases,including cardiovascular,cancer,diabetes mellitus,liver disease,Alzheimer’s disease,and androgenetic alopecia.In this study,we collected all the relevant experimental information on WDL from Scientific databases such as PubMed,Web of Science,Science Direct,and Google Scholar.Conclusively,WDL is recognized as a key anti-oxidant with both specific regulator and inhibitor of major drug targetable proteins in human chronic diseases and disorders.Hence,WDL as a novel therapeutic bioactive molecule is advised to explore further for relevant pharmacological activities.

Pharmacognostic Identification of Euphorbia maculata L.and Its Related Species

[Objectives]To make pharmacognostic identification of Euphorbia maculata L.and its related species.[Methods]The classical pharmacognostic identification method was adopted.[Results]The four main medicinal materials are very similar,the fluff can be seen in E.maculata and Euphorbia thymifolia L.,but not in Euphorbia prostrata Ait.and Euphorbia taihsiensis(Chaw et Koutnik)Oudejians;the tissue structure is basically the same;except for E.taihsiensis without non-glandular hairs,the powder has secretory cells,laticifers,cells,calcium oxalate crystals,fibers,vessels,and seed coat fragments.Through ultrasonic extraction with 80%ethanol,extraction with isobutanol,extending with chloroform∶ethyl acetate∶formic acid(5∶3∶0.3),developing color with 3%aluminum trichloride ethanol solution,under ultraviolet light(365 nm),the fluorescent spots of the same color appeared on the corresponding position of the chromatogram of the reference substance(quercetin,kaempferol).[Conclusions]It is not easy to distinguish the four medicinal materials by character identification and microscopic identification,while the thin-layer chromatography(TLC)is more reliable.

Discovery of the radio-protecting effect of Ecliptae Herba,its constituents and targeting p53-mediated apoptosis in vitro and in vivo

Radiation protection drugs are often accompanied by toxicity,even amifostine,which has been the dominant radio-protecting drug for nearly 30 years.Furthermore,there is no therapeutic drug for radiation-induced intestinal injury(RⅢ).This paper intends to find a safe and effective radioprotecting ingredient from natural sources.The radio-protecting effect of Ecliptae Herba(EHE) was discovered preliminarily by antioxidant experiments and the mouse survival rate after137Cs irradiation.EHE components and blood substances in vivo were identified through UPLC-Q-TOF.The correlation network of “natural components in EHE-constituents migrating to blood-targets-pathways” was established to predict the active components and pathways.The binding force between potential active components and targets was studied by molecular docking,and the mechanism was further analyzed by Western blotting,cellular thermal shift assay(CETSA),and Ch IP.Additionally,the expression levels of Lgr5,Axin2,Ki67,lysozyme,caspase-3,caspase-8,8-OHd G,and p53 in the small intestine of mice were detected.It was found for the first time that EHE is active in radiation protection and that luteolin is the material basis of this protection.Luteolin is a promising candidate for RⅢ.Luteolin can inhibit the p53 signaling pathway and regulate the BAX/BCL2 ratio in the process of apoptosis.Luteolin could also regulate the expression of multitarget proteins related to the same cell cycle.

墨旱莲总皂苷的质量标准研究

目的制定墨旱莲总皂苷的质量标准。方法测定5个不同产地的墨旱莲药材的水分、浸出物的含量;应用HPLC-ELSD法,建立墨旱莲中总皂苷含量的测定方法:对5个产地,共计10个批次药材进行含量测定。精密称取墨旱莲粉末1.0 g,置250 mL圆底烧瓶中,加入20 mL 70%乙醇回流提取2次,每次2 h。合并提取液,回收溶剂,提取物浸膏加甲醇溶解,过滤,定容至一定体积,测定含量。结果各产地药材含水量均符合《中华人民共和国药典》规定;药材醇溶性浸出物平均含量为22.74%,初步将浸出物含量标准定为不低于20%;10批药材墨旱莲总皂苷平均含量为0.56%,制定标准为不低于0.45%。结论2020版《中华人民共和国药典》一部中并没有对墨旱莲总皂苷类成分含量做出明确规定,该研究对墨旱莲总皂苷的质量标准做出补充,为墨旱莲皂苷成分的进一步开发利用奠定基础。

Chemical Constituents from Dried Aerial Parts of Eclipta prostrata

Abstract: Objective To study the constituents in the dried aerial parts of Eclipta prostrata. Methods The constituents wereisolated and purified by column chromatography and their structures were elucidated by spectroscopic methods (1D,2D NMR, UV, IR, and HRESI-TOF-MS) and chemical analyses. Results Eight compounds were isolated andidentified as 7-O-methylorobol-4′-O-β-D-glucopyranoside (1), 3′-hydroxybiochanin A (2), echinocystic acid28-O-β-D-glucopyranoside (3), ecliptasaponin A (4), eclalbasaponin I (5), eclalbasaponin IV (6), echinocystic acid(7), and 3-oxo-16α-hydroxy-olean-12-en-28-oic acid (8). Conclusion Compound 1 is a new compound andcompound 3 is obtained from this genus for the first time.

墨旱莲乙酸乙酯总提物对正常小鼠免疫功能的影响

目的 研究墨旱莲乙酸乙酯总提物 (EAEEP)对小鼠免疫功能的影响。方法 以大、中、小 3个剂量给药 ,考察 EAEEP对小鼠免疫脏器系数、碳粒廓清指数、迟发性变态反应、血清溶血素及脾抗体形成细胞等指标的影响。结果 EAEEP可显著降低小鼠的脾指数及碳粒廓清指数 ,抑制迟发性过敏反应 ,降低溶血素水平。EAEEP可显著提高小鼠的胸腺指数 ,对脾细胞抗体形成功能无显著影响。

墨旱莲的化学成分研究

目的研究墨旱莲的化学成分。方法应用溶剂法进行提取,硅胶、Sephadex LH-20等手段反复柱色谱分离、纯化,通过熔点、NMR等手段确定结构。结果从其乙醇提取物的石油醚、醋酸乙酯以及正丁醇萃取部分共得到10个化合物,分别为2,2′,5″,2″-三噻吩-5-羧酸(Ⅰ)、β-谷甾醇(Ⅱ)、芹菜素(Ⅲ)、槲皮素(Ⅳ)、木犀草素(Ⅴ)、蟛蜞菊内酯(Ⅵ)、去甲蟛蜞菊内酯(Ⅶ)、旱莲苷C(Ⅷ)、木犀草苷(Ⅸ)、蒙花苷(Ⅹ)。结论化合物Ⅰ为一新天然产物,Ⅹ为首次从墨旱莲中分离得到。

发根农杆菌RiT-DNA对墨旱莲的遗传转化

目的 建立墨旱莲 Eclipta prostrata的毛状根离体培养系统。方法 分别用发根农杆菌 A4,R16 0 1,ATCC15 8343种菌株感染墨旱莲的子叶外植体 ,获得毛状根 ,并筛选了优质株系 ;测定了毛状根的生长曲线 ;利用高压纸电泳法对毛状根进行 T- DNA转化的检测。结果 首次利用发根农杆菌 A 4,R16 0 1,ATCC15 8343种菌株成功地从墨旱莲中诱导出毛状根 ;经高压纸电泳检测 ,墨旱莲毛状根中含有甘露碱 ,表明 Ri质粒的 T- DNA已整合进毛状根中。

墨旱莲药材的高效液相指纹图谱及聚类分析

目的建立墨旱莲药材的指纹图谱研究方法并进行聚类分析。方法色谱柱:Betasil C18柱(250 mm×4.6 mm,5μm);流动相:体积分数为1%的磷酸水溶液和体积分数为1%的磷酸乙腈溶液,采用梯度洗脱;流速:1.0 mL.min-1;检测波长:330 nm;柱温:30℃。结果精密度与重复性试验中各共有峰的相对峰面积RSD小于5.0%,根据聚类分析结果,可将墨旱莲药材分为合格品和非合格品2类,合格品产地为浙江、江西、广东、武汉、江苏(苏州)、江苏(南京)、湖南、辽宁、河北、山西、北京、上海、安徽、海南,非合格品产地为河南、吉林、福建、山东、陕西(西安)。结论该指纹图谱研究及聚类分析方法可用于控制墨旱莲药材质量。

墨旱莲叶水提取物止血活性初探

[目的]探讨墨旱莲叶水提取物的止血和促凝血活性,为中草药在止血方面的应用提供依据。[方法]将不同剂量的墨旱莲提取物溶液与抗凝兔血混合,测定凝血时间。[结果]墨旱莲叶提取物为0.5%的最适浓度时具有最强促凝血活性,凝血时间为16 s,凝血时间缩短率为99.6%。[结论]墨旱莲叶水提取物有显著的止血和促凝血活性,具有较高的开发利用价值。

旱莲草对镉的富集特性研究

通过盆栽试验研究了不同镉浓度处理下旱莲草的镉富集特性。结果表明：随土壤镉浓度的增加,旱莲草生物量、株高、根长、叶绿素SPAD值、抗性系数都呈下降趋势,而根系和地上部镉含量呈增加的趋势。在土壤镉浓度为大于50 mg/kg时,旱莲草地上部分镉含量均大于镉超富集植物临界值（100 mg/kg）。在不同的土壤镉浓度处理下,旱莲草根系和地上部分镉富集系数（BCF）大于1,但转运系数（TF）小于1。旱莲草地上部分镉积累量最大值为38.06μg/株（土壤镉浓度为25 mg/kg）。小区试验研究表明,在土壤镉浓度为2.04～2.89 mg/kg时,旱莲草地上部分镉积累量为0.393～0.435 mg/m2。因此,旱莲草是一种镉富集植物,具有较好的镉富集能力,可用于农田镉污染土壤修复。

墨旱莲乙醇提取物动物致色素作用和对小鼠B16黑素瘤细胞黑素生成的影响

目的选择离体实验中对黑色素合成的关键酶—酪氨酸酶(tyrosinase,TYR)有激活作用的墨旱莲(旱莲草),研究其动物致色素作用和对小鼠B16黑素瘤细胞黑素生成及相关基因犤TYR、TYR相关蛋白(TRP-1/2)mRNA犦表达的影响。方法以棕色豚鼠为动物模型,用Schmorl法染色,计数含黑素的细胞;用多巴(DOPA)-氧化酶染色,计算每100个基底细胞中DOPA阳性细胞数。培养的小鼠B16黑素瘤细胞以四甲基偶氮唑盐比色(MTT)法、氢氧化钠(NaOH)法和体外氧化DOPA反应方法分别测定细胞的增殖活性、黑素生成量及TYR活性;逆转录-聚合酶链反应(RT-PCR)测定TYR及其TRP-1/2基因的表达。结果墨旱莲乙醇提取物可使豚鼠表皮基底层中含黑素颗粒细胞增多,使豚鼠表皮内DOPA阳性细胞增多(P<0.05);具有促进小鼠B16黑素瘤细胞黑素合成及TYR活性作用(P<0.05),对细胞TYR基因有上调作用,而对TRP-1/2mRNA的表达无明显作用。结论墨旱莲乙醇提取物具有促进黑素合成及上调酪氨酸酶基因表达的作用,对白癜风色素恢复有较好应用和开发的前景。

鳢肠不同部位抗骨质疏松活性及化学成分比较研究

为了比较鳢肠不同部位提取物的抗骨质疏松活性及总皂苷、总黄酮含量,采用MTT法和对硝基苯酚磷酸二钠法测定鳢肠不同部位乙醇提取物对UMR106细胞的增殖和碱性磷酸酶(ALP)活性;采用紫外分光光度法测定鳢肠不同部位总皂苷和总黄酮含量。结果表明,鳢肠根和叶提取物对UMR106细胞的增殖具有促进作用,浓度为50μg/mL时促增殖作用最强,增值率分别为7.7%和16.9%,茎提取物没有明显的作用;鳢肠根、茎、叶提取物对ALP活性均有促进作用,浓度为100μg/mL时促增殖作用最强,分别促进18.9%、20.1%和17.0%;鳢肠根、茎、叶中总皂苷含量分别为3.6%、4.0%和2.2%,总黄酮含量分别为1.4%、1.4%和3.4%。这些结果表明;鳢肠不同部位提取物均有抗骨质疏松活性,其不同部位总皂苷和总黄酮的含量差异较大。