基于肠道菌群-LPS/TLR4通路探讨野菊花提取物对“过食膏粱厚味”致高尿酸血症大鼠的作用

[目的]研究野菊花提取物(Chrysanthemi Indici Flos extract,CYM.E)对“过食膏粱厚味”致高尿酸血症(hyperuricemia,HUA)大鼠的治疗作用及机制。[方法]将64只大鼠随机分为正常对照组,模型对照组,别嘌醇10 mg·kg^(-1)组,苯溴马隆5 mg·kg^(-1)组,CYM.E15、30、60、90 mg·kg^(-1)组,每组8只。除正常对照组外,其余各组均灌胃脂肪乳剂,建立拟人“过食膏粱厚味”致HUA大鼠模型。实验期间测定血清尿酸(uric acid,UA)、总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-c)和低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-c)水平;酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)法测定血清白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和血浆脂多糖(lipopolysaccharide,LPS)水平;实时荧光定量聚合酶链式反应(Real-time quantitative polymerase chain reaction,qRT-PCR)法测定小肠中TNF-α、Toll样受体4(Toll like receptor 4,TLR4)的mRNA表达;免疫印迹法检测小肠中TLR4蛋白表达;应用16S rDNA测序技术,对肠道菌群进行操作分类单元(operational taxonomic units,OTU)分类,选择合适的OTU进行结构分析、LEfSe差异贡献分析等。[结果]CYM.E能显著降低模型大鼠血清UA、TC、LDL-c、IL-6和TNF-α水平;显著降低血浆LPS水平;同时显著下调小肠组织TNF-αmRNA、TLR4 mRNA表达和TLR4蛋白表达,改善肠道的微炎症状态;增加乳酸杆菌、双歧杆菌的丰度与种类,降低拟杆菌、大肠杆菌的丰度与种类。[结论]CYM.E具有降低血清UA水平、调节血脂、减低炎症损伤、减少LPS释放等作用,可能通过调节肠道微生态发挥作用。

Deletion of a 1,049 bp sequence from the 5′UTR upstream of the SiHEC3 gene induces a seed non-shattering mutation in sesame

Sesame is a labor intensive crop with limited mechanized harvesting mainly due to the seed shattering(SS)trait.In this study,we performed a genetic analysis of the seed-shattering resistance(SR)trait with a SR sesame mutant 12M07.Unlike the SS type,the parenchyma cells in the abscission zone of the 12M07 mutant are arranged loosely but adhere to the seed coat.Inheritance analysis of six generations derived from 12M07(SR)×Xiangcheng Dazibai(SS)showed that the SR trait is recessive and controlled by a single gene pair.Association mapping of the F2population with 888,619 variants(single-nucleotide polymorphisms(SNPs)and insertion-deletion(InDels))and 31,884 structural variations(SVs)determined that only SV12002 in the 5′upstream region of gene Sindi0765000(named SiHEC3)in Chr.3 was significantly associated with the SR trait.SiHEC3 encodes the bHLH transcription factor.A 1,049 bp deletion occurred in the 5′UTR of Sihec3 in 12M07.SiHEC3 is mainly expressed in developing placental tissues,with the expression peaking in capsules at 45 days after pollination.A dual-luciferase reporter assay in tobacco confirmed that the promoter activity of Sihec3 was reduced because of the deletion of the 1,049 bp promoter sequence.Protein–protein interaction network analysis showed that HEC3 is co-expressed with nine key proteins,such as SHATTERPROOF1(SHP1)and SEEDSTICK(STK)which participate in the secondary wall biosynthesis of the abscission layer in plants.The findings of this study show the important function of Sihec3corresponding with the SR trait and supply the genetic information for breeding new varieties that are amenable to mechanized harvesting in sesame and other crops.

Study on Extraction and Antioxidant Activity of Flavonoids from Dendranthema indicum

[Objectives] To determine the optimal extraction technology of flavonoids in Dendranthema indicum,and analyze the antioxidant activity of total flavonoids in D. indicum. [Methods]D. indicum was used as raw material to extract the total flavonoids by water extraction,alcohol extraction,freeze-thaw,microwave assisted extraction and ultrasonic extraction. [Results]Microwave assisted extraction had the highest extraction rate. Therefore,the microwave assisted extraction was selected as the main method,the ethanol aqueous solution was used as the solvent,and the single factor experiment was carried out with microwave power,ethanol volume fraction,solid-to-liquid ratio and microwave time as the variables. The optimal extraction process of flavonoids was microwave power 400 W,solid-to-liquid ratio 1 ∶ 30,extraction time60 s,ethanol volume fraction 70%,under this condition,the extraction yield was 10. 65 mg/g. The antioxidant activity of flavonoids in D. indicum was determined. The results showed that the highest scavenging rate of DPPH·free radicals was 47. 89%,and the highest scavenging rate of hydroxyl radicals was 98. 00%. [Conclusions]The flavonoids in D. indicum have excellent antioxidant activity. This study is expected to provide a basis for further development of D. indicum products.

野菊Dendranthema indicum(L.)Des Moul的大孢子发生和胚囊的发育

对菊科的野菊D.indicum进行了大孢子发生和胚囊发育的研究。单层的肥厚珠被起源于大孢子母细胞的基部附近。珠被内表皮早期就开始分化为珠被绒毡层。珠心属于瘦心式,单层的珠心表皮细胞在有效大孢子增大时开始解体。一个孢原细胞位于珠心表皮之下,直接起着大孢子母细胞的作用,大孢子四分体呈线形排列。胚囊的发育过程遵循正常的蓼型,但反足细胞核的产生有变化。胚胎学的资料不支持广义菊属Chrysanthemum L.的成立。著者在观察中发现因虫害形成胚囊吸器的例外情况。胚囊吸器的形成可能是因为胚珠不能满足胚囊发育所需营养而产生的一种生理适应。著者建议在今后的胚胎学研究中注意对珠被绒毡层进行观察和分类。

Typical HPLC Chromatogram of Petroleum Ether in Abutilon indicum(L.) Sweet from Different Habitats

[Objectives] To study the typical HPLC chromatogram of petroleum ether in Abutilon indicum (L.) Sweet from different habitats, in order to provide experimental basis for the quality control of A. indicum and the spectrum-efficacy relationship of petroleum ether.[Methods] The HPLC chromatogram of petroleum ether was obtained using Agilent SB-C 18 (4.6 mm×250 mm, 5 μm) and acetine-0.1% phosphoric acid solution for gradient elution at temperature of 30 ℃, flow rate of 1.0 mL/min and detection wavelength of 205 nm.[Results] The typical HPLC chromatogram of petroleum ether in A. indicum was preliminarily established, with 12 characteristic peaks. It was verified by A. indicum from different habitats, characterized by reproducibility and stability.[Conclusions] Typical HPLC chromatogram provides a stable new method for the quality control of petroleum ether in A. indicum . This study provides experimental basis for further study on the medicinal parts of A. indicum .

Combined Effects of Compost,Supraxone and Lambda-Super on Soil Microbial Activity under Pluvial Cultivation of Sesame(Sesamum indicum L.)in Burkina Faso

Sesame cultivation is strategic for the economy of Burkina Faso. Indeed, after cotton, sesame is the second most important agricultural product for exportation. However, its production is limited by plant diseases and pest attacks. Facing this situation, large amounts of pesticides are used to boost productivity. But, these chemicals have a negative impact on soil fertility. In this instance, the effects of Supraxone and Lambda-super in the presence of compost on soil microbial activity under rainfed sesame cultivation in a peasant environment were studied. The experiment design was a block of Fisher (BCR) comparing the non-treated parcel (TNT) to the parcel amended only with compost (C) and to the parcel amended with compost and treated with the pesticides (C + P), respectively. Soil samples were taken at depths of 0 - 20 cm before sowing at 41st and 81st days after sowing. Respiratory activity and soil microbial biomass were monitored respectively by respirometric tests and by the fumigation-incubation method. The results show that the addition of compost into the soil with or without supraxone treatment and Lambda-super resulted in an increase in microbial biomass and soil respiratory quotient. These results show that the provision of compost in sufficient quantity corrects the inhibitory effect of soil biological activity by pesticides.

A New Compound from the Bud of Chrysanthemum indicum L.

A new bicyclic spiroketone was isolated from the bud of Chrysanthemum indicum L. The chemical structure was elucidated as （1R, 9S, 10S）-10-hydroxyl-8 （2＇, 4＇-diynehexylidene）- 9-isovaleryloxy-2, 7-dioxaspiro [5, 4] decane based on the X-ray crystallography.

Genetic Diversity Assessment of Moroccan Sesame(Sesamum indicum L.)Populations Using Agro-morphological Traits

Genetic diversity of Moroccan sesame(Sesamum indicum L.)was analyzed for the first time using agro-morphological traits.Thirty-three sesame populations collected from Tadla region in Morocco were evaluated for 13 parameters in three contrasted environments,Afr-13,Afr-14 and Atj-14.Broad and significant variation among populations was observed only for three agro-morphologic traits,namely first capsule height(FCH),number of seeds per capsule(NSC)and thousand seeds weight(TSW).The respective overall averages were 71.82 cm(ranging from 65 cm to 77 cm),61.39(varying from 57 to 65)and 3.20 g(ranging from 3.0 g to 3.4 g).Environment had a highly significant effect on all studied parameters,except capsule size and oil content.The obtained mean values of parameters studied showed that environment Afr-14 was the most favorable for most of them,particularly seed yield(1.32 t/ha)and TSW(3.57 g).Significant and interesting correlations were observed between seed yield per plant(SYP)and other parameters.As relevant implication of this finding,number of capsules per plant(NCP)and NSC could be used as valuable selection criteria in sesame breeding program.However,limited and non-significant variation among sesame populations for most of parameters studied suggests that they are genetically very close and may be derived from a single cultivar or germplasm.Molecular characterization is needed to confirm or refute this hypothesis.Besides,when compared with sesame grown throughout the world,Moroccan sesame was found to be very promising,which open up the possibility to develop this crop in this country.

Effect of Microclimatic Modification on Growth and Yield of Sesame （Sesamum indicum L.） in a Semi-Desert Region of Northern Sudan

An investigation was carried out in the semi- desert region of northern Sudan, during 2000, to study the influence of different micrometeorological parameters on growth and yield of sesame under alley cropping system. Sesame, cv. Kenana-2, was grown in 6-m wide alleys formed by 3 year old Acacia ampliceps and Acacia stenophylla. Air temperature, relative humidity, wind speed, solar energy and shade behaviour were measured throughout the growing season. Irrigation was measured by water meter and soil moisture by gravimetric sampling at depths of 15, 30, 45 and 60 cm, in alley cropping and control plots. Sesame was evaluated for growth and yield performance in southern, central and northern parts of each alley. The results showed substantial reduction in wind runs, solar radiation and air temperature, while relative humidity was increased under alley conditions. Soil moisture content, especially in the upper 045 cm soil depth, was much higher in alley cropped plots than in the control plots. A. stenophylla resulted in the highest saving of irrigation water and considerably increased （40%） sesame seed yield. In contrast, sesame seed yield was reduced by 46% under A. ampliceps-alley cropping. Competition for light and water were highly responsible for variation in sesame seed yield in the different zones of the alley. A. stenophylla tree is, therefore, recommended in an alley cropping system in the semi-desert region of northern Sudan for its superiority in optimizing radiation and saving of irrigation water.

不同产地野菊花中黄酮类成分含量的HPLC分析

The content of linatin,luteolin,quercetin and apigenin in Dendranthema indicum(L.) Des Moul.flower from eleven locations was analyzed by HPLC.The results show that linatin and luteolin contents in samples are higher with a range of 0.0-13.3 and 0.5-2.1 mg·g-1,respectively,and suitable for quantitative determination,while apigenin and quercetin contents are trace and suitable for qualitative determination.The content of flavonoids is obviously affected by locations.No linatin is detected in samples from Jiangxi and Nanyang of He’nan.The content of linatin and luteolin is higher in samples from Chuzhou of Anhui,Nanjing of Jiangsu and Inner Mongolia.D.indicum flower from Chuzhou of Anhui is superior for having all four flavonoids and higher content of linatin and luteolin.

合成紫花茄皂苷诱导人肝癌BEL-7402细胞凋亡

目的:探讨合成紫花茄皂苷对BEL-7402细胞的增殖抑制及凋亡诱导作用。方法:采用酸性磷酸酶(APA)法、吖啶橙荧光染色法和流式细胞术检测合成紫花茄皂苷对人肝癌BEL-7402细胞增殖的影响以及诱导细胞凋亡的作用。利用蛋白印迹法检测药物作用后凋亡相关蛋白PARP的表达水平。结果:合成紫花茄皂苷对人肝癌BEL-7402细胞有明显的增殖抑制作用,其72h的IC50为4.2μg/ml。荧光显微镜下细胞呈现典型的凋亡形态。流式细胞术分析发现,细胞经皂苷作用6h、12h和24h后,凋亡率显著增加。蛋白印迹检测结果显示,合成紫花茄皂苷作用后肿瘤细胞内的PARP蛋白被剪切,Bcl-2蛋白的表达量下调。结论:合成紫花茄皂苷对人肝癌细胞的增殖抑制呈浓度相关性,具有诱导肿瘤细胞凋亡的作用,且这一作用可能与Bcl-2表达下调有关。

野菊花提取物对慢性支气管炎大鼠血清和支气管肺泡灌洗液中TNF-α、IL-6含量及肺组织病理形态学的影响

目的研究野菊花提取物对慢性支气管炎(CB)大鼠血清和支气管肺泡灌洗液(BALF)中TNF-α、IL-6含量以及肺组织病理形态学的影响,探讨其治疗CB的机制。方法制备野菊花提取物;Wistar大鼠随机分为空白对照组,CB模型组,喘息灵对照组,野菊花提取物低、中、高剂量组;气管内注入脂多糖建立CB大鼠模型,药物干预后,ELISA法测定大鼠血清及BALF中TNF-α及IL-6含量,观察肺组织的病理形态学变化。结果模型组大鼠血清及BALF中TNF-α及IL-6含量较空白对照组明显升高,肺组织发生明显病理损伤,野菊花提取物可显著降低上述异常升高的指标、改善肺组织的病理损伤。结论降低CB大鼠体内TNF-α、IL-6含量,改善肺组织的病理损伤,可能是野菊花提取物减轻CB炎症效应机制之一。

中药野菊花提取物与抗菌药联合对产ESBLs大肠杆菌抑制效果研究

试验旨在探讨野菊花提取物与抗菌药联合作用对产超广谱β-内酰胺酶(extended spectrumβ-lactamases,ESBLs)大肠杆菌的体外抑制效果,为野菊花的进一步开发利用提供理论依据。采用2倍微量稀释法和中西药联合诱导传代法测定野菊花提取物、抗菌药及联合作用后的最小抑菌浓度(minimal inhibitory concentration,MIC)。结果显示,野菊花水提物对该耐药大肠杆菌的MIC值为250mg/mL,且在野菊花水提物亚抑菌浓度(1/2 MIC)作用下进行中西药联合传代,其与头孢曲松钠、头孢噻呋钠、头孢噻肟钠、氟苯尼考、痢菌净、阿米卡星、林可霉素7种抗菌药联合作用后,抗菌药的MIC值明显降低。表明野菊花水提物能够显著降低产ESBLs大肠杆菌对部分抗菌药的敏感性。

芽期Al^(3+)胁迫对芝麻幼苗生长的影响及种质资源耐铝毒性评价

铝毒害是影响江西省红壤作物生长的重要因素之一,为了解铝对芝麻生长的毒害,发掘耐铝毒种质资源,用Al2(SO4)3·18H2O溶液模拟发芽期Al3+胁迫研究不同浓度Al3+对芝麻品种赣芝9号和赣芝12成苗及幼苗生长的影响。结果表明,Al3+浓度在0~100mg/L时,随着Al3+浓度的增加,成苗率、芽长、根长和苗鲜重均不断下降,且根长下降最为明显;而浓度在250~1 000mg/L时,芝麻难以成苗。Al3+浓度在50mg/L时,成苗率、芽长、根长和苗鲜重等测定指标均急剧下降,因此可认为50mg/L的Al3+浓度胁迫可以作为芝麻种质耐铝毒性筛选鉴定的参考浓度。进而在50mg/L Al3+胁迫下评价96份芝麻种质[含39个改良品种(系)和57份地方种质],可将它们聚为3类,即包含21份耐铝毒种质、37份中等耐铝毒种质和38份不耐铝毒种质;发掘出赣芝9号、金黄麻、武宁黑等21份耐铝毒种质。比较分析表明,地方种质资源的平均耐铝毒特性要优于改良品种(系),且除成苗率外,地方种质耐铝毒性的其他指标的变异系数和多样性指数均大于改良品种(系)。

芝麻耐湿性QTL定位及优异耐湿基因资源挖掘

【目的】芝麻是对湿害极其敏感的作物,湿害是影响中国芝麻生产发展和单产提高的主要障碍因素,然而,芝麻耐湿性分子生物学研究基础薄弱,迄今,国内外有关芝麻耐湿性基因定位的研究尚未见报道。利用重组自交系(RIL)群体进行芝麻耐湿性QTL定位,结合芝麻核心种质群体进行耐湿性相关分子标记研究,并挖掘优异耐湿基因资源。【方法】以高耐湿芝麻品种中芝13与极敏感种质宜阳白杂交后连续自交6代构建206个株系的RIL群体。利用113对多态性分子标记扫描RIL群体获得基因型数据,用MapMaker/EXP.3.0软件构建遗传连锁图谱。2009年和2010年在武汉和鄂州2地点通过人工淹水胁迫获得RIL群体盛花期湿害后正常株率和存活株率的表型数据,用Microsoft Excel 2010软件进行表型数据方差分析,用QTLNetwork 2.0软件基于复合区间作图法进行QTL定位,利用主效QTL紧密连锁的分子标记扫描核心种质群体,并结合耐湿性表型数据分析得到相关有效分子标记。通过盛花期耐湿性表型重复鉴定筛选,结合分子标记辅助选择,获得优异耐湿基因资源。【结果】构建的遗传连锁图谱全长592.4 cM,共有70个标记位点进入15个连锁群,标记间的平均距离为8.46 cM。共检测到与盛花期耐湿性相关的6个QTL位点,定位在第7、9、13和15连锁群上,分别解释5.67%—17.19%的表型变异,加性效应值2.7190—9.7302,贡献率最大的QTL为qWH10CHL09,加性效应3.9394,其增效等位基因来源于母本中芝13,SSR标记ZM428与其紧密连锁(遗传距离为0.7 cM)。标记ZM428在186份芝麻核心种质中验证结果表明,该标记2种基因型的资源间在耐湿表型上存在显著差异(P=0.0163),因此,标记ZM428可作为芝麻耐湿性分子辅助选择的有效标记。还挖掘出8份优异耐湿基因资源,湿害后其正常株率均>70%,存活株率均>80%。【结论】检测到6个芝麻耐湿性相关QTL,其中,贡献率最大的17.19%,获得1个有效分子标记,挖掘出8份优异耐湿基因资源。

木蝴蝶中黄酮类化合物的HPLC-ESI-MS分析

目的 建立木蝴蝶中黄酮类化合物的HPLC-ESI-MS分析方法,鉴定木蝴蝶中的黄酮类化合物.方法 色谱柱：Hypersil C18（250 mm×4.6 mm,5μm）;流动相：0.5%甲酸水溶液（A）、乙腈（B）,梯度洗脱;紫外检测波长范围：195～400 nm;采用正离子检测模式.结果 木蝴蝶中的6个黄酮类化合物获得了良好的分离与检测.通过与对照品的保留时间、正离子质谱比较鉴定和测定黄芩苷和白杨素的量,同时根据正离子质谱数据和文献数据分析推断了4个黄酮化合物.结论 本方法准确快速,适合木蝴蝶中黄酮类化合物的鉴定,可用于木蝴蝶原药材的质量控制.

合成紫花茄皂甙诱导人肝癌细胞株Bel-7402凋亡的作用机制

背景与目的:紫花茄(SolanumindicumL.)是一种有抗炎和治疗跌打损伤作用的传统中草药,含有丰富的结构独特的薯蓣皂甙,即紫花茄皂甙。我们的研究结果已证实,数种合成紫花茄皂甙有强抗肿瘤效果。本研究探讨合成紫花茄皂甙Ⅰ("-D-木吡喃糖基-(1→3)-[#-L-鼠李吡喃糖基-(1→2)]-"-D-半乳吡喃糖基薯蓣苷)的抗肿瘤机制。方法:以不同浓度紫花茄皂甙处理人肝癌细胞Bel-7402后,用酸性磷酸酶(acidphosphataseassay,APA)法测定增殖抑制率和IC50。用结晶紫染色显微镜观察细胞的形态学变化。用蛋白免疫印迹法检测凋亡过程中相关蛋白的变化。结果:紫花茄皂甙对Bel-7402细胞有明显的增殖抑制作用,在皂甙Ⅰ作用72h后的IC50为4.20μg/ml。光学显微镜下可见细胞密度降低,胞质中出现膜泡等形态学变化。蛋白免疫印迹检测结果显示,紫花茄皂甙Ⅰ作用后细胞质中的细胞色素c含量明显增加,Caspase-3被激活,细胞内的poly(ADR-ribose)polymerase(PARP)蛋白被剪切。结论:紫花茄皂甙对人肝癌细胞的增殖抑制呈浓度相关性,且通过线粒体依赖性通路诱导肿瘤细胞凋亡。

HPLC法测定神农香菊中蒙花苷的含量

目的:采用HPLC法测定神农香菊中蒙花苷的含量。方法:色谱柱为ZOBAX SB-C_(18)(250 mm×4.6 mm,5μm),流动相为甲醇-0.9%醋酸(56:44),流速为0.5 ml·min^(-1),柱温:30℃,检测波长为334 nm。结果:蒙花苷在1.2～12.0μg·ml^(-1)范围内有良好的线性关系,平均加样回收率为98.00%,RSD为1.61%。结论:本方法简便准确,重复性好,可用于神农香菊中蒙花苷的含量测定。

福田河白菊与其他菊花栽培品种HPLC指纹特征研究

目的比较不同产地各种菊花栽培品种的HPLC指纹特征,为构建福田河白菊的HPLC指纹图谱奠定基础。方法采用INERTSILC18柱,乙腈-水(含2%四氢呋喃,0.1%三氟醋酸)为流动相,梯度洗脱,体积流量1mL/min;柱温30℃,对不同菊花栽培品种以及同属其他相关种进行对比色谱分析。结果构建了菊花的HPLC指纹图谱模式,该模式显示不同产地菊花品种的HPLC图谱具有较高的相似性,有15个共有峰,其中4个为10强峰,2个为10强峰或次强峰。指纹数据反映了各品种菊花在地域和亲缘关系上的联系以及各自独特的特征。结论该色谱系统可以用于构建福田河白菊、湖北金菊等不同菊花品种的指纹图谱。

基于核糖体DNA的ITS序列和叶绿体trnT-trnL及trnL-trnF基因间区的菊花起源与中国菊属植物分子系统学研究(英文)

Several sequences were applied to resolve phylogenetic relationships within Dendranthema and clarify the origin of garden chrysanthemums including sequences of nrDNA ITS, trnT-trnL and trnL-trnF intergenic spacer regions in cpDNA. The relationships among the species are so close that the three sequences could only provide limited information. From the evidence presented, we suggest that: ① D.rhmobifolium be the chloroplast donor of D.vestitum (HN) with the resembling morphology and the same trnT/L IGS sequence. ② D.vestitum, a putative ancestor, may be not the chloroplast donor of garden chrysanthemums. D.lavand-ulifolium might be the chloroplast donor of the type population of D.indicum (HN) or the direct chloroplast donor of the ancient garden chrysanthemum cultivar. ③ D.zawaskii might be not the ancestor of garden chrysanthemums.