The diterpenoid esters from Euphorbia lathyris L. induce intestinal inflammation via LXRα/ABCA1-regulated lipid rafts and TLR4-mediated pathway

Background:Diterpenoid esters are considered to be the main toxic components and bioactive constituents of Euphorbia lathyris L.(EL).Euphorbia factors L1(EF1),L2(EF2),and L3(EF3),the main diterpenoid esters of EL,have been found to cause intestinal diarrhea and induce intestinal inflammation in mice.This research aimed to explore the effects of major diterpenoid esters from EL on intestinal inflammation,as well as to clarify their possible targets and molecular mechanisms in vivo and vitro.Methods:Caco-2 cells and BALB/c mice were intervened with EFL1,EFL2,and EFL3,respectively.The expressions of TLR4,NLRP3,NF-κB p65,LXRα,ABCA1,TNF-αand IL-1βwere measured by Real-time PCR and ELISA.Cholesterol efflux levels were examined using cholesterol efflux kit.Flow cytometry was applied to detect lipid rafts abundance.Confocal microscopy was applied to investigate co-localization of lipid rafts and TLR4.Results:Our results revealed that EFL1,EFL2,and EFL3 inhibited LXRα,ABCA1 expression,and cholesterol efflux,promoted colocalization of TLR4 and lipid rafts,and up-regulated TLR4,NLRP3,NF-κB p65,TNF-αand IL-1βexpressions.Conclusion:These findings reveal that the mechanisms by which EFL1,EFL2,and EFL3 induce intestinal inflammation may be associated with LXRα/ABCA1-regulated lipid rafts and TLR4-mediated pathways.

Occurrence and Control of Brown Patch in Energy Oil Plants Euphorbia lathyris L.

[ Objective] The purpose was to study the occurrence law and control effect of brown patch of E. lathyris. [ Method ] The pathogen of brown patch in E. lathyris was isolated and identified in Xingyi of Guizhou Province in 2008 -2009, and disease symptom observation and field fungicide efficacy test were conducted. [ Result ] The pathogen of brown patch in E. lathyris was fungus in Pseudoc ercospora sp., its suitable growth temperature was 20 -35 ℃, its occurrence peak was from June to September, and its field incidence were 30% -60%. Field efficacy test showed that 5 kinds of fungicides such as Thiophanate-methyl, Mancozeb, etc. had certain control effect on the disease, the relative control effect was 54. 3% -82.5%, in which mancozeb had better control effect, while Thiediazole copper had worse control effect. [Conclusion] The study would provide theoretical reference for further research on the cultivation of E. lathyris with hiah vield and aualitv and control of brown hatch.

Acid Value Determination and Pre-Esterification of Crude Euphorbia lathyris L. Oil

There were various problems in the determination of oil acid value of the national standard method, and this paper developed a low cost, simple and effective way to determine the acid value of oil and grease. Furthermore, the esterification of high acid Euphorbia lathyris L. oil (ELO) with methanol could be efficiently catalyzed by hydrochloric acid to produce biodiesel, and the influencing factors such as the amount of catalyst, reaction time, reaction temperature and molar ratio of oil to methanol were also studied. Under the optimized conditions with the oil to methanol molar ratio of 1:30 and a reaction temperature of 70℃, 95.8% oil conversion was obtained within 40 min in the presence of only 2.0 wt% of catalyst. Therefore, the low-cost non-edible Euphorbia lathyris L. oil as a raw material had good potential for the synthesis of biodiesel in industry.

Analysis and determination of diterpenoids in unprocessed and processed Euphorbia lathyris seeds by HPLC-ESI-MS

Euphorbia lathyris （Caper spurge） is a toxic and potent Chinese materia medica （T/PCMM）. This study sought a method for identifying five diterpenoids （Euphorbia factors LI-L3, L7a, and Ls） with the spectra of UV and mass, quantifying three diterpenoids L1, L2, and L8 in crude extracts of unprocessed and processed E. lathyris seeds by liquid chromatography/ electrospray ionization mass spectrometry （LC-ESI-MS）. The analysis was achieved on an Agilent Eclipse XDB-C18 column （4.6 mm× 150mm i.d., 5 μm） with an isocratic elution with a mobile phase consisting of water and acetonitrile at a flow rate of 0.25 mL/min at column temperature of 30 ℃ and UV detection was set at 272 nm. An ESI source was used with a positive ionization mode. The calibration curve was linear in the ranges of 9.9-79 μg/mL for Euphorbia factor Lb 3.8-30.5μg/mL for Euphorbia factor L2, and 1.0-20.6 μg/mL for Euphorbia factor LB. The average recoveries （n=6） of three diterpenoids were 98.39%, 91.10% and 96.94%, respectively, with RSD of 2.5%, 2.4% and 2.1%, respectively. The contents of the three diterpenoids in processed E. lathyris seeds were 3.435, 1.367 and 0.286 mg/g, respectively, which decreased more sharply than those in unprocessed E. lathyris seeds which were 4.915, 1.944 and 0.425 mg/g, respectively. The method is simple, accurate, reliable and reproducible, and it can be applied to control the quality of unprocessed and processed E. lathyris seeds.

小飞扬草(Euphorbia thymifolia L.)中镉的亚细胞分布及化学形态

采用差速离心法和逐步提取法,分析Cd超积累植物小飞扬草根、茎、叶中Cd的亚细胞分布和化学形态。培养液中Cd浓度5mg/L和10mg/L条件下,在小飞扬草根、茎、叶中,细胞壁和细胞膜是Cd主要的结合场所,含量分别占总量的47.1%～49.0%、39.7%～41.4%、42.9%～56.2%,其次是胞液组分,含量分别占37.5%～41.4%、28.2%～40.7%、35.2%～46.8%,细胞器组分中Cd含量较少。Cd在小飞扬草根、茎、叶中均以氯化钠提取态为主,含量分别占70.5%～84.8%、72.4%～83.0%、50.0%～74.8%。根部各提取态含量依次为CNaCl>CHAC>CEtOH>CHCl>CW>CR,茎、叶中各提取态含量依次为CNaCl>CHCl>CHAC>CEtOH>CW>CR。在小飞扬草中,Cd与细胞壁和细胞膜中的果胶酸和蛋白质等物质结合固定,而进入细胞的Cd大部分与有机酸络合而隔离于液泡内,这可能是小飞扬草忍耐并超积累Cd的机制。

Antioxidant activity and phytochemical screening of the methanol extracts of Euphorbia hirta L

Objective:To assess antioxidant activities of different parts of Euphorbia hirta（E.hirta）,and to search for new sources of safe and inexpensive antioxidants.Methods:Samples of leaves,stems, flowers and roots from E.hirta were tested for total phenolic content,and flavonoids content and in vitro antioxidant activity by diphenyl-1-picrylhydrazyl（DPPH） assay and reducing power was measured using cyanoferrate method.Results:The leaves extract exhibited a maximum DPPH scavenging activity of（72.96±0.78）%followed by the flowers,roots and stems whose scavenging activities were（52.45±0.66）%,（48.59±0.97）%,and（44.42±0.94）%,respectively.The standard butylated hydroxytoluene（BHT） was（75.13±0.75）%.The IC50,for leaves,flowers,roots,stems and BHT were 0.803,0.972,0.989,1.358 and 0.794 mg/mL,respectively.The reducing power of the leaves extract was comparable with that of ascorbic acid and found to be dose dependent. Leaves extract had the highest total phenolic content[（206.17±1.95） mg GAE/g],followed by flowers,roots and stems extracts which were（117.08±3.10） mg GAE/g,（83.15±1.19） mg GAE/g,and （65.70±1.72） mg GAE/g,respectively.On the other hand,total flavonoids content also from leave had the highest value[（37.970±0.003） mg CEQ/g],followed by flowers,roots and stems extracts which were（35.200±0.002） mg CEQ/g,（24.350±0.006） mg CEQ/g,and（24.120±0.004） mg CEQ/ g,respectively.HPTLC bioautography analysis of phenolic and antioxidant substance revealed phenolic compounds.Phytochemical screening of E.hirta leaf extract revealed the presence of reducing sugars,terpenoids,alkaloids,steroids,tannins,flavanoids and phenolic compounds. Conclusions:These results suggeste that E.hirta have strong antioxidant potential.Further study is necessary for isolation and characterization of the active antioxidant agents,which can be used to treat various oxidative stress-related diseases.

A new aryl glycoside from Euphorbia helioscopia L.

A new aryl glycoside, 3″-O-galloyl-benzyl-O-α-L-rharnnopyranosyl-（1 → 6）-β-D-glucopyranoside, was isolated from Euphorbia helioscopia L., and its structure was elucidated on the basis of various spectroscopic data analysis.

A transmission electron microscopy study of the diversity of Candida albicans cells induced by Euphorbia hirta L.leaf extract in vitro

Objective:To determine the major changes in the microstructure of Candida albicans(C. albicans) after treatment with Euphorbia hirta(E.hirta) L.leaf extract.Methods:Transmission electron microscopy was used to study the ultrastructural changes caused by E.hirta extract on C. albicans cells al various exposure time.Results:It was found that the main abnormalities were the alterations in morphology,lysis and complete collapse of the yeast cells after 36 h of exposure to the extract.Whereas the control cultures showed a typical morphology of Candida with a uniform central density,typically structured nucleus,and a cytoplasm with several elements of endomembrane system and enveloped by a regular,intact cell wall.Conclusions:The significant antifungal activity shown by this methanol extract of E.hirta L.suggests its potential against infections caused by C.albicans.The extract may be developed as an anticandidal agent.

A new lathyrane diterpene glycoside from Euphorbia helioscopia L.

A new lathyrane diterpene glycoside,named 3β,7β,15β-trihydroxy-14-oxolathyra-5E,12E-dienyl-16-O-β-D-glucopyranoside, was isolated from Euphorbia helioscopia L.Its structure was established by spectroscopic techniques including 2D NMR.

Configuration Confirmation of Euphorbia Factor L_1 by Single-crystal X-ray Diffraction

A 6,（17）-epoxylathyrol diterpenoid （（2S＊3S＊4R＊5R＊6S＊9S＊11S＊15R＊）-5,15-diacetoxy-3- phenylacetoxy-14-oxolathyra-6（17）,（12E）-diene-6（17）-epoxide） was isolated from the seeds of Euhporbia lathyris L. Its configuration was puzzled because of the incomplete X-ray results reported before. In this work, the atom connectivity and configuration were confirmed by single-crystal X-ray diffraction together with ESI-MS, ^1H-, and ^13C-NMR spectroscopy. The compound crystallizes in monoclinic, space group P21 with a = 11.386（1）, b = 8.2839（7）, c = 17.192（2） A, β = 108.305（2）°, Z = 2, V = 1539.5（2）A^3, C32H40O8, Mr = 552.64, Dc = 1.192 g/m^3, /7（000） = 592,μ（MoKα） = 0.085 mm^-l, T = 293（2） K, the final R = 0.0398 and wR = 0.0950 for 2057 observed reflections with Ⅰ 〉 2α（Ⅰ）. The molecule shows a tricyclic terpenoid skeleton, consisting of fused five-, eleven- and three-membered rings. The configuration at C（5） is R^＊ and that at C（6） S^＊.

Antimicrobial activity of Tunisian Euphorbia paralias L.

Objective: To examine the potential antimicrobial activity of Euphorbia paralias L. (Euphorbiaeae) leaves and stems extracts. Methods: The antimicrobial activity was tested against six microbial strains:Escherichia coli ATCC 8739, Bacillus subtilis ATCC 6633, Salmonella enterica CIP 8039, Staphy-lococcus aureus ATCC 6538, Pseudomonas aeruginosa ATCC 9027 and Candida albicans ATCC 90028 by two different methods, the disk method and the dilution method. Results: Our results showed the important antimicrobial activity of the chloroform extract of the stems towards the majority of the strains by using both methods. Bacillus subtilis was the most sensitive strain (MIC=MBC=15μg/mL). Conclusion: Thus, some extracts of Euphorbia paralias can be used in the treatment of infectious diseases caused by microbes.

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.

NaCl胁迫对续随子离子吸收及荧光特性的影响

以续随子为材料采用温室盆栽法,研究了NaCl胁迫处理对其生长发育、离子分布和叶绿素荧光特性的影响。结果表明:(1)25mmol.L-1NaCl胁迫时续随子幼苗的鲜重、干重和株高比对照略有下降,但不显著;随着NaCl浓度的继续增加,三指标均显著降低。(2)盐胁迫影响植物组织的离子分布,Na+浓度增加时,K+、Ca2+和Mg2+浓度均持续下降,K+/Na+、Ca2+/Na+、Mg2+/Na+也急剧下降。(3)NaCl胁迫下,续随子幼苗叶片的初始荧光(Fo)、最大荧光(Fm)、PSⅡ实际光化学效率均降低。但在0～25mmol.L-1NaCl浓度下,Fo、Fm与对照差异不显著。

复方泽漆散对小鼠Lewis肺癌形态学影响

目的:探讨复方泽漆散对小鼠Lewis肺癌的作用。方法:本研究以移植性C57BL/6小鼠Lewis肺癌为动物模型,观察复方泽漆散对小鼠Lewis肺癌的抑制作用,及其对肿瘤细胞病理形态学的影响。结果:复方泽漆散治疗小鼠Lewis肺癌,能够抑制肿瘤细胞生长,稳定病灶;中剂量组能诱导肿瘤细胞凋亡;本方不同浓度组皆可导致肿瘤细胞坏死,线粒体在形态学上皆有明显改变,或肿胀,或空泡变,或消失。结论:复方泽漆散对小鼠Lewis肺癌具有一定治疗作用,其作用机理可能与促进肿瘤细胞坏死和凋亡,影响线粒体形态和功能有关。

黔产千金子HPLC指纹图谱研究

文章采用国家药典委员会《中药色谱指纹图谱相似度评价系统》2004年A版对所得HPLC图谱进行相关分析,建立了黔产千金子药材的指纹图谱并确定了8个特征峰,该方法简便、可靠、重现性好,可用于对黔产千金子药材的质量评价。

续随子ElFAD2基因的序列及表达特性分析

为了研究油酸脱氢酶(FAD2)基因 ElFAD 2对续随子( Euphorbia lathyris L.)中不饱和脂肪酸合成的调控作用,该研究在续随子转录组数据的基础上经筛选获得 ElFAD 2基因序列,并对其序列及表达特性进行分析。序列分析结果显示, ElFAD 2基因全长1 907 bp,ORF长1 152 bp,共编码383个氨基酸,包含有典型的脂肪酸去饱和酶结构域。续随子ElFAD2蛋白理论等电点为8.08,属于稳定蛋白,包含4个跨膜区和3个保守的组氨酸簇。基于FAD2的系统发育分析表明,续随子与同科植物乌桕( Triadica sebifera L.)的亲缘关系最近。荧光定量PCR分析发现, ElFAD 2基因在不同器官中均有表达,且在花后15 d的种子中表达量最高,在叶与花后30 d及45 d种子中的表达量相当,而在根、茎、花中的表达量最低。该研究结果为深入探讨续随子 ElFAD 2基因的生物学功能提供了基础数据,也为解析续随子种子中脂肪酸合成的分子机制奠定了基础。

续随子ElDGAT1基因的克隆、序列分析及表达特性

二酰甘油酰基转移酶(Diacylglycerol acyltransferase,DGAT)是植物三酰甘油(TAG)合成最后一步反应的关键酶和限速酶,属于酰基转移酶超家族。为探究续随子(Euphorbia lathyrisL.)中二酰甘油酰基转移酶1(DGAT1)对于调控种子油脂合成的作用机理,依据续随子转录组数据,采用RT-PCR技术分离和鉴定出一个DGAT1基因(命名为ElDGAT1),运用生物信息学方法对获得的序列进行分析,通过qPCR技术研究ElDGAT1基因在续随子不同器官中的表达特性。结果表明:从续随子中克隆获得ElDGAT1基因的cDNA序列,编码区长为1530bp,共编码509个氨基酸;预测ElDGAT1蛋白定位于内质网上,且为疏水性膜结合蛋白;其二级结构以α-螺旋和无规则卷曲为主,编码的蛋白含有9个典型的跨膜螺旋区。基于DGAT1蛋白的系统发育分析表明:续随子与乌桕的亲缘关系最近,与橡胶树和木薯的同源性次之。qPCR检测结果发现,ElDGAT1基因在续随子根中表达量最低,种子中的表达量相对较高,在花后15d、30d、45d的表达量呈递增趋势。研究结果为进一步分析续随子种子油脂合成积累的调控机理及ElDGAT1基因的生物学功能提供依据。

UPLC-MS测定千金子提取物的活性成分及其抗氧化性研究

目的:鉴定千金子乙醇提取物化学组成,探究其抗氧化活性,筛选活性化合物。方法:基于超高效液相色谱-飞行时间质谱(UPLC-TOF-MS),数据库检索和文献比对等手段对其活性成分进行分析;采用DPPH和ABTS自由基清除法、β-胡萝卜素漂白法对提取物进行抗氧化活性评价。结果:共鉴定出17个化合物,包括青蒿亭、槲皮素-3-O-葡萄糖醛酸苷、山奈酚-3-O-葡萄糖醛酸苷、蔓荆子黄酮4个黄酮类化合物,瑞香素和双七叶内酯2个香豆素类化合物;千金子乙醇提取物对DPPH自由基和ABTS自由基的清除能力远大于BHT;对β-胡萝卜素的漂白作用弱于BHT。结论:千金子乙醇提取物中含有较丰富的水溶性黄酮类化合物和香豆素类化合物,表现出一定的抗氧化活性。

续随子ElPDAT1基因生物信息学及表达特性分析

磷脂:二酰甘油酰基转移酶1(PDAT1)是不依赖于脂酰-CoA催化二酰甘油(DAG)合成三酰甘油(TAG)的关键酶,在种子油脂代谢过程中起关键作用。为了研究ElPDAT1基因在续随子种子油脂合成过程中的调控作用,对ElPDAT1进行了生物信息学分析,并采用qRT-PCR技术研究了该基因在续随子不同器官中的表达特性。结果表明,ElPDAT1基因c DNA全长2 968 bp,其中,开放阅读框为2 037 bp,共编码678个氨基酸,预测等电点和相对分子质量分别是5.94,75.38 ku。ElPDAT1蛋白二级结构的主要结构元件是α-螺旋(34.81%)和无规则卷曲(43.95%)。ElPDAT1基因系统进化树分析表明,续随子与同科植物蓖麻、木薯、麻疯树的亲缘关系较近。qRT-PCR研究发现,ElPDAT1基因在各个器官中均有表达,且在种子中的表达量明显高于其他器官;ElPDAT1基因在种子发育中期表达量达到最大,其表达量为叶的5.82倍,说明ElPDAT1基因可能在续随子种子油脂合成过程中起调控作用。研究结果可为揭示该基因的生物学功能及全面解析续随子油脂合成机制提供科学依据。

续随子转录因子ElWRI1的鉴定及功能分析

续随子(Euphorbia lathyris L.)种子可积累大量油脂且富含油酸,WRINKLED1(WRI1)为AP2/EREBP类转录因子,参与调控脂肪酸的合成及种子发育等生命过程。本研究通过对续随子的全基因组鉴定,获得了11个续随子ElWRI1转录因子基因,命名为ElWRI1-1~ElWRI1-11。结果显示,它们均具有两个AP2结构域;其编码蛋白均属于不稳定亲水蛋白,无跨膜结构;亚细胞定位预测显示所有WRI1蛋白均位于细胞核。表达分析结果表明,ElWRI1-6与ElWRI1-8在种子发育时期的表达量较高,且后者的表达模式与种子油脂积累模式一致。在本氏烟草(Nicotiana benthamiana L.)叶片中异源过表达ElWRI1-8,叶片中总油脂含量增加,且油酸含量显著上升。研究结果表明ElWRI1-8可能是调控续随子种子油脂和油酸生物合成的一个重要转录因子。