Effect of Elephantopus scaber L.Extract on Acute Pleurisy in Rats

[Objectives]To study the anti-inflammatory effect of Elephantopus scaber L.extract on acute pleurisy induced by carrageenan in rats,and to explore its anti-inflammatory mechanism.[Methods]The active sites of E.scaber L.were extracted by ethanol reflux method.The extracts of different concentrations of E.scaber L.were used as the study object,and dexamethasone was used as the positive control drug.The anti-inflammatory effects of E.scaber L.extracts were studied by measuring the levels of malondialdehyde(MDA),prostaglandin E_(2)(PGE_(2)),tumor necrosis factor(TNF-α),interleukin-1β(IL-1β)in pleural fluid and serum nitric oxide(NO),MDA,PEG_(2),TNF-α,IL-1βin rats with acute pleurisy induced by carrageenan.[Results]E.scaber L.extracts in three doses could reduce the levels of inflammatory factors in pleural fluid and serum,and inhibit acute pleurisy in rats.It was speculated that the anti-inflammatory mechanism was related to the inhibition of the release of inflammatory factors and the antioxidant effect of extracts of three doses of E.scaber L.[Conclusions]This experiment provides a basis for the development and application of E.scaber L.

Phenolic compounds from Elephantopus scaber L.

Eight compounds,(+)-syringaresinol(1),apigenin(2),3,5-dimethoxy-4-hydxyphenylpropanol(3),3,4,5-trimethoxy benzoic acid,(4)ferulaic acid(5),trans-p-hydroxycinnamic methyl ester(6),4-hydroxy-3-methoxybenz-oic acid(7)and p-methoxy benzoic acid(8)were isolated from the 70%ethanol extract of the whole herb of Elephantopus scaber L.The structures of all compounds were identified by spectral data analysis.All compounds were obtained for the first time from Elephantopus scaber L..

Virtual screening of active ingredients,potential targets and pathways from Elephantopus scaber L.against liver cancer based on network pharmacology

Elephantopus scaber L.,belonging to Elephantopus plant of Compositae,is widely used as the common Chinese herbal medicine in folk.According to the phytochemical investigations,E.scaber is used to treat several cancers,anemia and diarrhea on account of the property of the complex components and multi-targets.However,the underlying molecular mechanisms of herb remain unclear.In this report,network pharmacology method is used to analyze the relation among the small-molecule compounds of E.scaber,target proteins,biological pathways and liver cancer.The potential active compounds from the E.scaber,potential target proteins and the key pathways were predicted by network pharmacology models.We believe that our present study is useful to find novel lead compounds for liver cancer.

Research review on the main chemical constituents and pharmacological effects of Elephantopus scaber L.

Elephantopus scaber L.belongs to Elephantopus plant of Compositae,is widely used as the common Chinese herbal medicine in folk.Phytochemical investigations of the E.scaber have resulted in the isolation of triterpennoids,sesquiterpene lactones,sesquiterpene phenols,steroids,flavones,anthraquinones,peptides,lipid compounds and aryl compounds.It possesses the effects of antibacterial,clearing away heat and detoxification,detumescence and diuresis,and is used for treating lung cancer,cervical cancer,liver cancer,nasopharyngeal cancer,breast cancer,anemia and diarrhea.The research progress in the chemical constituents ofE scaber and pharmacological effects are reviewed for better explore and utilize theE scaber resource.

Isolation and identification of chemical constituents from Elephantopus scaber Linn.

Elephantopus scaber L.,as a popular Chinese medicinal herb of the Compositae family,is widely distributed in America,Africa and Asia.In the present study,five compounds were isolated from the 70%EtOH extract of the whole herb from Elephantopus scaber L.,including two lignans and three flavones.Their chemical structures were identified based on NMR spectroscopic analysis and comparison with the data reported in literature.It is noteworthy that compounds 1-5 were isolated from Elephantopus scaber L.for the first time and their chemotaxonomic significance was discussed.

Antineoplastic effects of deoxyelephantopin,a sesquiterpene lactone from Elephantopusscaber, on lung adenocarcinoma (A549) cells

OBJECTIVE： Deoxyelephantopin, a sesquiterpene lactone from Elephantopus scaber, showed inhibition of the growth of various tumor cells in vitro. In the present study, we investigated the cytotoxicity and apoptosis-inducing capacity of deoxyelephantopin on lung adenocarcinoma （A549） cells. METHODS： The cytotoxic effect of deoxyelephantopin on A549 cells and normal lymphocytes was evaluated using 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay and 50% inhibitory concentration （IC50） value was determined. The self-renewal and proliferating potential of A549 cells after treatment with deoxyelephantopin were examined by colony formation assay. Cellular morphology of deoxyelephantopin-treated cells was observed using phase- contrast microscopy. The induction of apoptosis was evaluated using acddine orange and ethidium bromide staining, Hoechst 33342 staining, terminal deoxynucleotidyl transferase-mediated dUTP biotin nick end-labeling （TUNEL） assay, DNA fragmentation analysis and Annexin V-fluorescein isothiocyanate staining by flow cytometry. Activation of caspases was detected using fluorogenic substrate specific to caspases 2, 3, 8 and 9 and flow cytometric analysis. The total cellular DNA content and expression of cleaved poly （ADP-ribose） polymerase was also analyzed. RESULTS： Deoxyelephantopin exhibited cytotoxicity to A549 cells （IC50 = 12.287 μg/mL）, however, there was no toxicity towards normal human lymphocytes. Deoxyelephantopin suppressed the colony-forming ability of A549 cells in a dose-dependent manner. Acridine orange, ethidium bromide and Hoechst 33342 staining showed cell shrinkage, chromosomal condensation and nuclear fragmentation, indicating induction of apoptosis. Deoxyelephantopin increased apoptosis of A549 cells, as evidenced by more TUNEL-positive cells. DNA fragmentation and Annexin V staining revealed late-stage apoptotic cell population. Deoxyelephantopin inhibited A549 cell growth by cell cycle arrest at G2/M phase and induced apoptosis through both extrinsic and intrinsic pathways. CONCLUSION： These results suggest that deoxyelephantopin has great potential as a new chemotherapeutic agent to be developed further for the treatment of lung cancer.

Sesquiterpene Lactones from Elephantopus scaber

A new germacranolide sesquiterpene lactone, isoscabertopin, was isolated from Elephantopus scaber together with the known scabertopin. Their structures were determined by spectroscopic methods.

Effect of Elephantopus scaber Linn on community acquired pneumonia through TLR4/NF-κB signaling pathway

Objective:To observe the effect of Elephantopus scaber Linn on community-acquired pneumonia and its relationship with the expression of TLR4/NF-κB pathway and the downstream inflammatory cytokines IL-6,IL-8,TNF-α.and to explore its molecular mechanism for the treatment of community-acquired pneumonia,so as to provide new ideas and theoretical basis for the treatment of community-acquired pneumonia.Methods:140 patients with community-acquired pneumonia were randomly divided into observation group and control group,with 70 cases in each group.Observation group:Elephantopus scaber Linn+NS atomized inhalation+intravenous injection of ceftazidine;Control group:intravenous injection of ceftazidine;Meanwhile,70 healthy subjects were selected as the healthy control group.The expressions of TLR4,NF-kB,IL-6,IL-8 and TNF-αin serum of observation group,control group before and after treatment and healthy control group were detected by enzyme linked immunosorbent assay(ELISA),and the efficacy of Elephantopus scaber Linn in the treatment of community acquired pneumonia was observed.Results:Compared with the control group,the observation group was significantly better than the control group in recovery rate,total effective rate,number of days for the disappearance of symptoms such as fever,cough,expectoration,with statistical significance(P<0.05);Compared with the healthy control group,the serum TLR4,NF-κB,IL-6,IL-8,TNF-αwere significantly different(P<0.05)between the observation group and the control group;Serum TLR4,NF-κB,IL-6,IL-8,TNF-αwere significantly different(P<0.05)in the observation group before and after treatment with Elephantopus scaber Linn;Compared with the control group after treatment,there were significantly different(P<0.05)in serum TLR4,NF-κB,IL-6,IL-8 and TNF-αin the observation group after treatment.Conclusion:Serum TLR4,NF-κB,IL-6,IL-8,TNF-αwere highly expressed in patients with community-acquired pneumonia,Elephantopus scaber Linn can decreased the expression of TLR4 and NF-κB in blood,and its downstream inflammatory factors also decreased significantly.There were significantly different(P<0.05)in blood TLR4 and NF-κB between the observation group and the control group after treatment.It is suggested that Elephantopus scaber Linn may inhibit TLR4/NF-kB signal pathway to exert anti-inflammatory effect.Elephantopus scaber Linn has a definite effect and can significantly improve the clinical symptoms,and effectively shorten the treatment cycle.

Virtual screening of the active compounds and potential targets of Elephantopus tomentosus L. for the treatment of liver cancer based on a network pharmacology approach

Liver cancer is the leading cause of cancer-related death.As the clinical treatment for this disease is not satisfactory,it is urgent to discover new medications to treat liver cancer.Elephantopus tomentosus L.is a traditional whole grass herb,which contains sesquiterpene lactones,triterpenoids,aldehyde caffeic with good anti-tumor effect.In this study,molecular docking and network analysis were carried out to identify the active compounds,and predict the potential targets of E.tomentosus for the treatment of liver cancer.These attempts may provide insights for finding novel lead compounds for the treatment of liver cancer.

基于TLR4/NF-κB信号通路探讨地胆头干预社区获得性肺炎机制研究及临床观察

目的:观察地胆头对社区获得性肺炎的疗效,探讨其与TLR4/NF-κB信号转导通路和下游炎性因子IL-6、IL-8、TNF-α表达的关系,以期为社区获得性肺炎的治疗提供新的思路和理论依据。方法:将140例社区获得性肺炎患者,随机分为观察组及对照组各70例,观察组:地胆头液+生理盐水雾化吸入+头孢他啶针静脉注射;对照组:头孢他啶针静脉注射治疗,选择同期70名健康体检者为健康对照组;用酶联免疫吸附法(ELISA)检测观察组、对照组治疗前后及健康对照组血清中TLR4、NF-κB、IL-6、IL-8、TNF-α表达,并对照观察地胆头治疗社区获得性肺炎的效果。结果:与对照组相比,观察组痊愈率、总有效率、发热、咳嗽、咳痰症状消失天数明显优于对照组,有统计学意义(P<0.05);与健康对照相比,观察组及对照组血清TLR4、NF-κB、IL-6、IL-8、TNF-α有明显差异(P<0.05);地胆头治疗前后,观察组血清TLR4、NF-κB、IL-6、IL-8、TNF-α有明显差异(P<0.05);与对照组治疗后比,观察组治疗后血清TLR4、NF-κB、IL-6、IL-8、TNF-α有明显差异(P<0.05)。结论:社区获得性肺炎患者血清TLR4、NF-κB、IL-6、IL-8、TNF-α呈高表达状态,地胆头液降低血液中TLR4、NF-κB的表达,其下游炎症因子也明显下降,观察组、对照组治疗后血液中TLR4、NF-κB存在明显差异(P<0.05),说明地胆头可能是通过抑制TLR4/NF-κB信号通路来发挥抗炎作用;地胆头治疗社区获得性肺炎疗效确切,能显著改善临床症状,有效缩短治疗周期。

Apelin/APJ在人支气管上皮细胞炎症反应中的作用及地胆头对其影响的研究

目的:探讨Apelin/APJ系统在急性肺损伤(ALI)细胞模型的作用,研究中药地胆头(草)对支气管上皮细胞Apelin/APJ系统及炎症介质的影响。方法:依据对人支气管上皮细胞(16HBE)施加条件不同(n=8),分为正常对照组(阴性对照组),模型组(LPS组),阳性对照组(LPS+Apelin-13组),低、中、高地胆头组[LPS+ESE(20μg/ml)组、LPS+ESE(40μg/ml)组、LPS+ESE(80μg/ml)组]。激光共聚焦扫描显微镜法检测16HBE细胞及炎症模型Apelin/APJ表达与定位;CCK8法检测地胆头对LPS诱导的细胞损伤的影响;ELISA法测定各组支气管上皮细胞上清液炎症因子TNF-α、IL-1β、IL-6的分泌水平;RT-PCR及Western blot法检测16HBE中Apelin/APJ的转录及表达。结果:Apelin、APJ均在16HBE细胞膜上表达;10μg/ml LPS干预后Apelin/APJ细胞膜表达明显增多(P<0.05),IL-1β、IL-6和TNF-α细胞因子水平明显升高(P<0.05),Apelin-13+LPS组中16HBE的Apelin/APJ表达上调,支气管上皮细胞炎症反应减轻(P<0.05),地胆头预处理可明显提高Apelin/APJ表达,抑制LPS诱导的炎症细胞因子的产生(P<0.05)。结论:Apelin/APJ系统在急性肺损伤/急性呼吸窘迫综合征(ALI/ARDS)过程中被激活,对抗炎症反应。地胆头通过上调16HBE细胞Apelin/APJ系统,减轻炎症反应,从而发挥抗炎作用。

地胆草中羽扇豆醇的提取及采用反相HPLC法测定其含量

我们采用化学提取法从地胆草中提取了羽扇豆醇,并建立了反相-HPLC法测定地胆草中羽扇豆醇含量的方法,为地胆草的质量分析和评价提供科学依据。采用了Wondasil C18柱(4.6 mm × 150 mm, 5 μm),柱温为30?C,甲醇-水(9:1)为流动相,在流速1.0 mL?min?1和紫外检测波长206 nm下进样20 μL对地胆草石油醚提取物进行HPLC分析。结果发现:羽扇豆醇在0.25~5.0 μg?mL?1 (r = 0.9998)呈良好的线性关系,平均加样回收率为98.1% (RSD 1.87%)。本方法简便,重复性好,为评价地胆草药材的质量提供科学依据。

基于氧化应激和TLR4/NF-κB炎症信号通路研究地胆头治疗慢性阻塞性肺疾病急性加重疗效

目的观察地胆头治疗慢性阻塞性肺疾病急性加重(AECOPD)患者的疗效及对氧化应激和Toll样受体4(TLR4)/核转录因子κB(NF-κB)炎症信号通路及下游炎性介质表达的影响,以期为AECOPD防治提供新依据。方法选择2018年10月—2019年6月在海南医学院第一附属医院住院治疗的AECOPD患者160例,按随机数字表法分成观察组与对照组各80例,对照组给予抗菌、平喘等常规治疗,观察组在对照组治疗基础上给予地胆头液雾化吸入,2组均治疗14 d。比较2组治疗效果,检测2组治疗前后血清炎症相关指标[TLR4、NF-κB、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)]、氧化应激指标[活性氧自由基(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)]水平和肺功能[第1秒用力呼气容积(FEV_(1))、FEV_(1)/用力肺活量(FVC)]。结果观察组总有效率为95.0%(76/80),对照组为81.2%(65/80),观察组明显高于对照组(P<0.05)。2组治疗后血清TLR4、IL-6、IL-8、TNF-α、ROS、MDA水平及观察组NF-κB水平均明显降低(P均<0.05),且观察组各指标水平均明显低于对照组(P均<0.05);2组治疗后血清SOD水平及FEV_(1)、FEV_(1)/FVC均明显增高(P均<0.05),且观察组各指标均明显高于对照组(P均<0.05)。结论地胆头可通过抑制氧化应激及TLR4/NF-κB炎症信号通路,减少炎症因子释放,从而提高AECOPD治疗效果,明显改善肺功能。

Interventional study of neutrophil elastase-mediated pathway for biliary head to interfere with high secretion of airway mucus in patients with chronic obstructive pulmonary disease

Objective:To observe the effect of Elephantopus scaber Linn on chronic obstructive pulmonary disease(COPD),to investigate its effect on the MUC5AC hypersecretion of airway mucus mediated by neutrophil elastin in patients with acute exacerbation of COPD,and to observe its effect on lung function(FEV1,FEV1/FVC),so as to provide new ideas and theoretical basis for the prevention and treatment of COPD.Methods:160 patients with acute exacerbation of COPD were randomly divided into observation group and control group with 80 cases each according to the numerical table method.The observation group was treated with aerosol inhalation of Elephantopus scaber Linn on the basis of conventional treatment.Both groups were treated for 2 weeks.During the same period,80 healthy patients were selected as the healthy control group.The expression of NE and MUC5AC in serum of observation group,control group and healthy control group before and after treatment was detected by enzymelinked immunosorbent assay(ELISA).The changes of lung function(FEV1,FEV1/FVC)after treatment in the observation group and the control group were compared and analyzed,and the efficacy of Elephantopus scaber Linn in patients with acute exacerbation of copd was observed.Results:Compared with the control group,the total effective rate of the observation group was significantly better than that of the control group,with statistical significance(P<0.05).Compared with the healthy control group,there were significant differences in serum NE and MUC5AC between the observation group and the control group(P<0.05).Serum NE,MUC5AC,FEV1 and FEV1/FVC were significantly different in the observation group before and after treatment(P<0.05).Serum NE,MUC5AC,FEV1 and FEV1/FVC were significantly different in the observation group before and after treatment(P<0.05).Compared with the control group after treatment,there were significant differences in serum NE and MUC5AC in the observation group after treatment(P<0.05),and the increase of FEV1/FVC and FEV1 was significantly better than that of the control group after treatment(P<0.05).Conclusion:Elephantopus scaber Linn can significantly improve the clinical treatment effect of patients with acute exacerbation of COPD,improve lung function and airway mucus hypersecretion.It can effectively inhibit the expression of NE and MUC5AC in the blood.There were significant differences in serum NE and MUC5AC between the observation group and the control group after treatment(P<0.05),showing a positive correlation,suggesting that inhibition of MUC5AC secretion by Elephantopus scaber Linn in patients with COPD may be related to NE-mediated pathways.

UPLC-MS/MS同时测定地胆草中黄酮类成分含量

地胆草中黄酮成分经乙醇溶液提取后,通过大孔树脂纯化,以Waters BEH C18柱分离。以1%甲酸的水溶液和1%甲酸的乙腈溶液为流动相,流速为0.2m L·min^(-1),柱温为30℃,进样体积为1μL。采用UPLC/MS/MS电喷雾电离,多反应监测模式检测,以保留时间和子离子比定性,外标法定量。在该色谱条件下,地胆草中的5种黄酮类成分能得到较好的分离并检测。结果表明,地胆草中木犀草素平均含量为8.75g·kg^(^(-1)),山奈酚平均含量为0.86g·kg^(-1),儿茶素平均含量为4.29g·kg^(-1),芹菜素平均含量为2.43g·kg^(-1),芦丁平均含量为10.71g·kg^(-1)。地胆草中黄酮类化合物种类丰富,含量较高,该研究为地胆草黄酮类化合物的含量测定及中草药地胆草的进一步开发应用提供理论依据。

双标多测法同时测定地胆草中11种成分的含量

目的建立双标多测法同时测定地胆草中新绿原酸、绿原酸、隐绿原酸、木犀草苷-7-O-葡萄糖醛酸苷、异绿原酸B、异绿原酸A、异绿原酸C、去氧地胆草素、异去氧地胆草素、异地胆草种内酯、地胆草种内酯的含量。方法分析采用Waters Symmetry C_(18)、Phenomenex C_(18)、Agilent ZORBAX Eclipse Plus C_(18)色谱柱(4.6 mm×250 mm,5.0μm);流动相乙腈-0.1%磷酸,梯度洗脱;体积流量1.0 mL/min;柱温35℃;检测波长220、326 nm。以绿原酸为内标,计算新绿原酸、隐绿原酸、木犀草苷-7-O-葡萄糖醛酸苷、异绿原酸B、异绿原酸A、异绿原酸C相对校正因子;以异去氧地胆草素为内标,计算去氧地胆草素、地胆草种内酯、异地胆草种内酯相对校正因子,测定其含量。再进行聚类分析、主成分分析、正交偏最小二乘法-判别分析。结果11种成分在各自范围内线性关系良好(r≥0.9990),平均加样回收率95.3%~103.4%,RSD 0.32%~3.45%。双标多测法所得结果与外标法接近。异绿原酸A、异绿原酸C、异绿原酸B、绿原酸、木犀草苷-7-O-葡萄糖醛酸苷、隐绿原酸为质量差异成分。结论该方法稳定可靠,可用于地胆草的质量控制。

地胆草化学成分的研究

采用硅胶、葡聚糖凝胶SephadexLH-20和十八烷基硅烷键合硅胶(ODS)等多种色谱技术,对地胆草根部的化学成分进行分离纯化,从地胆草醇提物的乙酸乙酯萃取部位中分离了9个化合物,通过理化性质和波谱数据鉴定化合物1-9的化学结构分别为:大黄素甲醚(1),异香草酸(2),香豆酸(3),对羟基苯甲酸(4),阿魏酸(5),3-甲氧基-4-羟基-桂皮醛(6),豆甾醇-3-O-β-D-葡萄糖(7),豆甾醇(8),吲唑(9).化合物1、2、4、5、6、9为首次从该种植物中分离得到,其中化合物9为一新的天然产物.

地胆草的化学成分研究

对地胆草全草的化学成分进行研究。采用硅胶柱色谱、SephadexLH-20以及半制备型高效液相色谱技术进行分离纯化，通过1D、2DNMR，MS等方法鉴定化合物的结构。结果从地胆草95％乙醇提取物中分离并鉴定了9个化合物，分别为1-[（2R＊，3．S＊）-3-ethoxy-2，3-dihydro-6-hydroxy-2一（1-methylethenyl）-1-benzofuran-5-y1]ethanone（1）、7-hydroxy-6．acetyl-2．methylchromone（2）、matriisobenzofrrran（3）、桦木酸（betulinicacid，4）、木犀草素（1uteolin，S）、木犀草素-7-D-β-D-葡萄糖苷（1uteolin-7-01B-D—ghcoside，6）、对香豆酸（P-coumaricacid，7）、3，4-二羟基苯甲醛（3，4-dihydroxybenzaldehyde，8）、豆甾醇-3-O-β-D-葡萄糖苷（stigmasterol-3-O-β-D—ghcoside，9）。其中化合物1—3均为首次从该属植物中分离得到，化合物3在国内未见从其它植物中分离报道。

地胆草三萜成分的研究

目的分离鉴定地胆草的化学成分,为PC-12细胞活性筛选提供样品。方法用体积分数80%工业酒精提取,硅胶柱色谱分离纯化,根据理化性质和光谱分析确定结构。结果分得8个三萜类成分,分别为无羁萜酮(friedlin,E-6),表无羁萜醇(epifriedlinol,E-7),羽扇豆醇(lupeol,E-8),羽扇豆醇乙酸酯(lupeol acetate,E-9),乌苏酸(ursolic acid,E-10),乌苏-12-烯-3β-十七酸酯(ursa-12-ene-3β-heptadecanoate,E-11),桦木酸(betulinic acid,E-12),30-羟基羽扇豆醇(30-hydroxylupeol,E-13)。结论化合物E-11为一个新的天然产物;化合物E-6,E-10,E-12,E-13为首次从该属植物中得到。

遮光与定植方式对地胆草生长及产量的影响

目的：了解地胆草在不同光照条件下的生长与产量表现,为地胆草的人工种植提供参考。方法：以全光照的露地栽培为对照,对遮光率分别为42%、50%、60%、69%、85%的遮光处理下穴播、苗植两种方式种植的地胆草生长状况及产量进行了研究。结果：适度遮光促进地胆草株高、叶片数、根系长度的增加并提高根冠比;随遮光率的增加,地胆草由定植到抽薹盛期、开花盛期所需时间减少;单位面积地胆草种子产量、全草产量、根头产量均为全光照的最高,并随遮光率增加而下降,各处理单位面积种子产量在8.46-31.10 kg/667 m^2之间,全草干品产量在327.28-800.95 kg/667 m^2之间,根头干品产量在30.65-70.72 kg/667 m^2之间。结论：地胆草对光照的适应能力较强,属偏喜阳,但可耐受较大程度的荫蔽;人工种植地胆草以穴播方式为好,在热带经济林下间种建议荫蔽度不超过60%为宜。