GC–MS analysis of bioactive compounds present in different extracts of an endemic plant Broussonetia luzonica (Blanco) (Moraceae) leaves

Objective: To investigate and characterize the chemical composition of the different crude extracts from the leaves of Broussonetia luzonica(Blanco)(Moraceae)(B. luzonica), an endemic plant in the Philippines.Methods: The air dried leaves were powdered and subjected to selective sequential extraction using solvents of increasing polarity through percolation, namely, n-hexane,ethyl acetate and methanol to obtain three different extracts. Then, each of the extracts was further subjected to gas chromatography–mass spectrometry.Results: Qualitative determination of the different biologically active compounds from crude extracts of B. luzonica using gas chromatography–mass spectrometry revealed different types of high and low molecular weight chemical entities with varying quantities present in each of the extracts. These chemical compounds are considered biologically and pharmacologically important. Furthermore, the three different extracts possess unique physicochemical characteristics which may be attributed to the compounds naturally present in significant quantities in the leaves of B. luzonica.Conclusions: The three extracts possess major bioactive compounds that were identified and characterized spectroscopically. Thus, identification of different biologically active compounds in the extracts of B. luzonica leaves warrants further biological and pharmacological studies.

Manual thinning increases fruit size and sugar content of Citrus reticulata Blanco and affects hormone synthesis and sugar transporter activity

Manual fruit thinning(MFT)in fruit trees has been previously shown to increase fruit size and enhance fruit quality,but the effect of MFT on Ponkan(Citrus reticulata Blanco)and the underlying mechanisms remain poorly understood.In this study,efforts were made to elucidate how MFT influences the fruit quality of Ponkan.The results showed that MFT substantially increased fruit size and elevated fruit total soluble solids in comparison with the fruit from the unthinned trees(used as control).Expression analyses demonstrated that m RNA abundance of three important sugar transporter genes,including CrSUT1,CrSTP1 and CrTMT1,was evidently elevated in the flesh of thinned fruit when compared with those of the control.In addition,MFT prominently up-regulated the transcript levels of various auxin and gibberellin(GA)biosynthesis and signaling genes,including CrYUC6,CrAUX/IAA,CrGA20ox1 and CrGA3ox1.Concurrently,the contents of endogenous IAA and GA3,measured at 90 d after fruit thinning,were notably elevated in the fruit from trees with the thinning treatment relative to the control,although no difference was detected in the two groups before the thinning manipulation.Taken together,these results indicate that manual fruit thinning could greatly improve fruit quality,which may be attributed to promoting fruit expansion due to the increased auxin levels and expediting sugar accumulation through the up-regulation of sugar transporter genes.

Lupeol:An antioxidant triterpene in Ficus pseudopalma Blanco(Moraceae)

Objective:To assess the antioxidant activity of Ficus pseudopalma Blanco(Moraceae)(F.pseudopalma) and characterize the active components present in it.Methods:Column chromatography of crude ethanol leaf extract of F.pseudopalma was performed and seven fractions were obtained,labeled as Ft.F2,F3,F4.F5,F6,F7.DPPH.FRAP,Criess,Kenton and superoxide radical scavenging assays were performed to assess the antioxidant ability of the fractions.Thin layer chromatography(TLC),high performance liquid chromatography and Fourier transfer infrared spectroscopy(FTIR) were performed to identify and characterize the bioactive component present in each fractions of F.pseudopalma.Results:DPPH and FRAP assay showed that F5.F6 and F7 exhibited the good proton accepting ability and reducing power as compared to the other fractions.All fractions exhibited a good nitric oxide radical scavenging activity wherein F1,F2 and F3 showed the highest inhibition.However,all of the fractions exhibited a stimulatory activity on hydroxyl and superoxide radicals.Lupeol matched one of the spots on the thin layer chromatography chromatogram of the fractions.Linear gradient high performance liquid chromatography and spiking of lupeol with the fraction revealed the presence of 5.84 mg/L lupeol in F6.Infrared spectra of the fractions revealed the presence of C-C,OH,aromatic C=C and C=O groups.Conclusions:The identified lupeol in F.pseudopalma may be responsible for the exhibited antioxidant property of the plant.Furthermore,knowing the antioxidant capability of the plant,F.pseudopalma can be developed into products which can help prevent the occurrence of oxidative stress related diseases.

In Vitro and In Silico Studies of Lunacridine from Lunasia Amara Blanco as Anticancer

Lunasia amara Blanco is a famous plant in South Sulawesi. It was used largely by local people as antibacteria and aphrodisiac. Quinoline alkaloid lunacridine was known as the active principle from Lunasia amara Blanco. Its activity was reported as a DNA Intercalating Topoisomerase II inhibitor. In this study, we have isolated and assayed the cytotoxic activity of lunacridine on P388 murine leukemia cells by MTT colorimetric assay （in vitro）. Lunacridine showed the less cytotoxic activity with the IC50 of 39.52 μg/mL. With the aim to explore the structural determinants responsible for this activity, molecular docking study have been carried out with DNA model using AutoDock 4.0 software with various total of energy evaluations （in silico）. The best docking reached at the total energy evaluations of 2.5 × 107 with the binding free energy of-6.63 kcal/mol. Analysis of the docking results was in accordance with the ability of lunacridine to intercalate between base pairs of DNA. Cytotoxic activity of lunacridine is less probably due to low affinity and molecular interaction. Therefore this study suggests to design and to develop lunacridine as a lead compound for anticancer drug.

陈皮精油对脂多糖诱导的RAW264.7细胞炎症的干预作用

基于脂多糖(lipopolysaccharide,LPS)诱导的RAW264.7细胞模型研究陈皮精油抗炎作用。通过Griess法检测一氧化氮(nitric oxide,NO)分泌水平,采用Real-time PCR法检测细胞中肿瘤坏死因子(tumor necrosis factorα,TNF-α)、白细胞介素6(interleukin-6,IL-6)、白细胞介素1β(interleukin-1β,IL-1β)和诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、环氧化酶2(cyclooxygenase 2,COX-2)mRNA的表达水平。结果显示,陈皮精油显著降低LPS诱导的RAW264.7细胞释放NO的含量,抑制iNOS、COX-2、IL-1β、IL-6和TNF-αmRNA的表达。综上所述,陈皮精油通过抑制炎症因子的分泌和表达,有效缓解由LPS诱导RAW264.7细胞炎症反应。

陈皮多糖提取工艺及其抗氧化活性研究

该研究以广陈皮为原料,利用超声波辅助提取法提取陈皮多糖,采用单因素试验优化陈皮多糖提取工艺并对其抗氧化活性进行研究,建立以葡萄糖为标准品,采用苯酚-硫酸法测定广陈皮多糖含量的定量分析方法。采用正交试验以多糖得率为指标优化工艺参数:料液比为1∶45(g/mL),提取时间为10 min,超声功率为350 W,提取温度为65℃,在此条件下,陈皮多糖得率高达40.06%。结果表明,广陈皮多糖对1,1-二苯基-2-三硝基苯肼自由基(DPPH·)的最大清除率为85.19%,对羟基自由基(·OH)的最大清除率为63.42%。

潮州柑果皮多糖对凝固型酸奶冷藏期品质的影响

以潮州柑果皮多糖为原料,通过评价凝固型酸奶的理化和感官特性,探讨潮州柑果皮多糖添加量对酸奶冷藏过程中品质的影响。研究结果表明:在7 d的冷藏期内,添加潮州柑果皮多糖可提高酸奶的可溶性固形物含量、黏度、持水力和悬浮稳定性,降低酸奶的乳清析出率;当多糖添加量为2.0%时,酸奶的综合品质最佳。冷藏第7天,添加2.0%多糖的酸奶pH值为4.12,可滴定酸度为96.61°T;相比对照组,可溶性固形物含量、黏度、持水力和悬浮稳定性分别提高了9.08%、40.59%、25.41%和145.83%,乳清析出率降低了55.37%。另外,相比其他组,添加1.5%和2.0%潮州柑果皮多糖的酸奶感官评分更高。因此,潮州柑果皮多糖可作为一种天然稳定剂可用于提高冷藏期凝固型酸奶品质。

橘叶化学成分及其抗炎活性研究

目的研究橘Citrus reticulata Blanco叶化学成分及其抗炎活性。方法橘叶85%乙醇提取物采用硅胶、D101大孔树脂、MCI、ODS、Sephadex LH-20进行分离纯化,根据理化性质及波谱数据鉴定所得化合物的结构。采用Griess法测定化合物对脂多糖诱导RAW 264.7细胞产生NO的抑制活性。建立角叉菜胶致小鼠足肿胀炎症模型,检测IL-1β、TNF-α水平。结果从中分离得到12个化合物,分别鉴定为川陈皮素(1)、橘红素(2)、5-去甲川陈皮素(3)、5,4′-二羟基-6,7,8,3′-四甲氧基黄酮(4)、5-羟基-7,8,3′,4′-四甲氧基黄酮(5)、3,5,6,7,8,3′,4′-七甲氧基黄酮(6)、橙皮素(7)、5-羟基-6,7,3′,4′-四甲氧基黄酮(8)、β-香树脂醇(9)、豆甾-5-烯-3β-醇(10)、对羟基苯甲醛(11)、香草醛(12)。化合物1~4、6~7、10、12对脂多糖诱导RAW 264.7细胞释放NO有较强抑制活性,IC_(50)值分别为(25.21±2.10)、(37.77±0.50)、(38.19±1.58)、(21.89±1.73)、(43.81±1.18)、(47.98±2.55)、(41.23±1.11)、(43.80±1.43)μmol/mL。化合物2~3可降低IL-1β、TNF-α水平(P<0.05,P<0.01)。结论化合物6~7、9为首次从该植物中分离得到。化合物1~4、8有较强的体外抗炎活性,化合物2~3有明显的体内抗炎活性。

广藿香^(60)Co-γ射线辐射诱变植株的筛选及评价

为应对广藿香药材需求量急速增加、无突出优点的品种且品种单一的情况,解决广藿香道地药材缺失及种质退化、种质来源混乱、易遭受病害虫害等问题,本研究通过辐照诱变育种,设置梯度剂量的^(60)Co-γ射线,对愈伤组织、丛生芽、生根苗3种不同生长阶段的广藿香进行辐照处理,观测再生植株外观形态,筛选出变异植株,测量其农艺性状和生理生化指标,采用气相色谱-质谱联用技术(GC-MS)测定其主要挥发性成分广藿香酮及百秋李醇含量。结果表明:愈伤组织和丛生芽对^(60)Co-γ射线较为敏感,死亡率较高,生根苗对^(60)Co-γ射线较为耐受,死亡率较低,最佳诱变育种的方式是采用100Gy^(60)Co-γ射线辐照处理生根苗,其诱变率高达10%。本研究最终获得29株广藿香变异植株,分别为三叶轮生、异型叶、深裂叶、狭叶、卷叶、下卷叶和紫色叶7种形态类型植株,通过比较分析渗透调节物质(可溶性糖、可溶性蛋白和游离脯氨酸)、抗氧化物质(丙二醛、过氧化物酶、过氧化氢酶)、广藿香酮和百秋李醇含量,最终筛选得到15株具有潜力植株,分别为:抗逆且高有效成分植株SY7和ZY1;对环境敏感且高有效成分植株SY15、SY18和XY1;抗逆植株SY2、SY6、SY19、YY2、YY9和XJY1;高有效成分植株SY10和SY17;“酮型”广藿香植株SY3和JY1。其中SY7和ZY1为双重优良特性新种质,2株“酮型”广藿香植株对于解决道地药材缺失及其种质退化等问题具有应用价值。本研究采用^(60)Co-γ射线辐照诱变方法选育的广藿香变异植株,具有较强抗逆性、高有效成分等优点,为广藿香诱变育种以及新品种选育提供成熟方法和潜在种质。

基于矿物元素指纹图谱鉴定陈皮产地的方法研究

采用电感耦合等离子体质谱法测定了新会、广西、湖南3地42批陈皮样品中51种矿物元素的含量,运用方差分析和偏最小二乘法-判别分析(PLS-DA)研究了不同产地陈皮中矿物元素的差别,以筛选得到的9种元素为溯源指标,将不同数量级矿物元素含量进行分类和系数缩放后,构建了陈皮中矿物元素的指纹图谱,并基于各产地矿物元素含量的平均值构建了新会、广西、湖南产地陈皮中矿物元素的标准指纹图谱。通过采用SPSS20.0计算18批新会陈皮矿物元素指纹图谱与其标准指纹图谱的相似度,确定新会陈皮矿物元素指纹图谱的相似度阈值为0.941,基于此阈值建立了新会陈皮的鉴别模型。采用24批样品对所建立的鉴别模型进行准确性验证,正确率为91.6%。该方法操作简单直观,满足实验分析要求,可为陈皮的产地溯源提供参考。

基于AHP-熵权法与Box-Behnken优化陈皮挥发油提取及包合工艺

目的:筛选陈皮挥发油最佳提取、包合工艺。方法:运用Box-Behnken响应面设计法优化陈皮挥发油提取工艺;采用层次分析法(AHP)-熵权法和L 9(34)正交实验,以羟丙基-β-环糊精与挥发油的比例、包合温度、包合时间为影响因素,以含油率、包合物得率和包合率为评价指标,优选陈皮挥发油最佳包合工艺。结果:挥发油最佳提取工艺为液料比为10∶1、浸泡时间为1.5 h、提取时间为4 h。挥发油最佳包合工艺为羟丙基-β-环糊精与挥发油的比例为6∶1,包合温度30℃,搅拌时间为1 h。结论:优化得到的陈皮挥发油提取及包合工艺,稳定可靠,能够为工业化生产提供依据。

狮头柑皮油成分分析及在卷烟加香中的应用

为开发天然烟用香料,采用压榨法提取了狮头柑皮油,利用顶空固相微萃取-气相色谱-质谱方法鉴定了其主要挥发性成分,并进行了卷烟加香评价。结果表明,狮头柑皮油中含有49种挥发性成分,加入到卷烟中能够显著改善卷烟的品质,增强柑橘果甜香的香韵特征,提高烟气的丰富甜润感,表明狮头柑皮油是可以利用的一种新型烟用香原料。

基于网络药理学和实验验证探讨臻呼畅组合物改善急性肺损伤的作用机制

该研究利用TCMSP数据库和Swiss Target Prediction平台获取臻呼畅组合物(简称SSD)中各中药的活性成分和靶点,利用GeneCards、DisGeNET、GEO数据库获取急性肺损伤相关靶点。通过Cytoscape 3.9.1软件、STRING数据库构建中药-活性成分-靶点-疾病网络、PPI网络,筛选出核心成分和核心靶点。利用David数据库对关键靶点进行GO和KEGG分析。应用AutoDock Vina 1.2.3软件对核心成分和核心靶点进行分子对接,PyMOL 2.6.0软件进行可视化处理。采用脂多糖(LPS)诱导的急性肺损伤(Acute lung injury,ALI)大鼠模型,验证SSD改善急性肺损伤的作用。结果表明:筛选出SSD活性成分30个,改善肺损伤相关靶点9个,其中核心成分主要为β-谷甾醇(β-sitosterol)、亚油酸乙酯(mandenol)、山奈酚(kaempferol)、26-O-β-D-glucopyranosyl-3β,26-dihydroxy-5-cholesten-16,22-dioxo-3-O-α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranoside_qt,核心靶点主要为白细胞介素-6(Interleukin-6,IL-6)、肿瘤坏死因子(Tumor necrosis factor,TNF)、环加氧酶-2(prostaglandin-endoperoxide synthase 2,PTGS2)、Jun原癌基因(Jun proto-oncogene,JUN)、纤溶酶原激活物抑制因子-1(Serpin E1,SERPINE1)。GO和KEGG分析结果显示,SSD主要通过白细胞介素-8(Interleukin-8,IL-8)生成的正向调节、白细胞与血管内皮细胞粘附、脂多糖反应、血管内皮生长因子产生,与蛋白酶结合、IL-6受体结合、转录调控区序列特异性DNA结合,调控糖尿病并发症中的AGE-RAGE信号通路、TNF信号通路等协同发挥改善急性肺损伤的作用。分子对接证明3个核心成分与5个核心靶点均结合良好。动物实验结果表明中高剂量的臻呼畅组合物可显著降低ALI大鼠肺泡灌洗液、肺组织中的炎症因子TNF-α、IL-1β和IL-10水平,病理学观察其可减轻肺组织损伤。该研究初步揭示了臻呼畅组合物改善急性肺损伤的多成分、多靶点、多途径的协同作用机制,为SSD应用于肺部健康的保健及ALI的预防提供理论依据。

橘园生草栽培的研究进展

柑橘(Citrus reticulata Blanco)生草栽培技术作为一种土壤管理技术,具有提高土壤肥力,改善土壤生态环境,提高果实产量及品质的功能。综述了生草对橘园病虫害、生态环境、土壤物理性质、土壤pH、土壤养分、土壤有机质、土壤微生物和土壤酶等果树不同生长环境因素的影响,并探讨了生草促进果树生长,提高果实产量和品质的作用,进而提高柑橘的生产性能和经济效益的作用方式。

湖北部分地区柑橘农药残留情况调查与分析

采用超高效液相色谱-串联质谱联用仪和气相色谱-串联质谱联用仪对湖北省宜昌市区、丹江口市、秭归县和兴山县的柑橘(Citrus reticulata Blanco)样品农药残留情况进行定量检测和分析。结果表明,柑橘样品农药残留的检出率高达91.9%,其中,禁用农药检出率为1.6%;限用农药检出率为9.5%;允许使用农药检出率为68.3%;未登记使用农药检出率为15.9%。农药使用情况较为普遍,不过由于禁、限用农药控制程度较好,因此柑橘质量整体安全性较高。

湖北柑橘主产区主要病虫害种类及发生期调查

为了解湖北省柑橘(Citrus reticulata Blanco)主产区主要病虫害种类及发生特点,2017—2018年连续两年采用实地调查,并采集标本进行种类鉴定。结果表明,主要病害有11种,以柑橘炭疽病、沙皮病、绿斑病(青苔病)、根腐病为害最严重;主要害虫有19种,以柑橘红蜘蛛、潜叶蛾、柑橘大实蝇为害最严重。

春季防虫网温湿度变化对砂糖橘春梢生长的影响

探讨了防虫网覆盖栽培技术对网室温湿度和砂糖橘(Citrus reticulata Blanco cv.Shatangju)春梢生长的影响。结果显示,40目透光防虫网对1-4月份(春季)砂糖橘春梢的生长具有促进作用;同时比较了防虫网内外温湿度变化对粉虱类、红蜘蛛类、潜叶蛾类数量的影响,认为防虫网对砂糖橘春梢生长的促进作用可能是通过对柑橘害虫和网室温湿度的调控这两个方面来实现的。

广藿香的基因序列与挥发油化学型的相关性分析

目的 探讨“南药”广藿香Pogostemoncablin (Blanco)Benth .不同产地间的叶绿体和核基因组的基因型与挥发油化学型的关系 ,为广藿香道地性品质评价、规范化种植提供分子依据。方法 用PCR直接测序技术对广藿香6个产地样本的叶绿体matK基因和核 18SrRNA基因核苷酸序列进行测序分析研究。结果 广藿香 6个样本的matK基因序列长均为 12 4 5bp ,编码 4 15个氨基酸成熟酶。 18SrRNA基因序列长为 180 3～ 180 5bp。根据排序比较 ,广藿香 6个样本间的matK基因序列存在 4 7个变异位点 ,18SrRNA基因存在 17个变异位点 ,非加权组平均法构建的系统分支树表明广藿香基因序列分化与其产地、所含挥发油化学变异类型呈良好的相关性。结论 结合挥发油分析数据 ,基因测序分析技术可作为广藿香道地性品质评价方法这一以及规范化种植过程关键技术“物种鉴定”

1961～2009年湖北省柑橘冻害等级及其特征分析

利用湖北省70个气象站点1961～2009年冬季气温、降水资料,分析了湖北省区域柑橘(Citrus reticulata Blanco)越冬期极端低温、低温最长持续时间、降水最长持续时间和降水量的时空变化特征,并以此为基础提出柑橘综合冻害指数和冻害等级的对应关系。结果表明,自1961年以来,湖北省冬季极端最低气温以每10年0.62℃的速率升高,全省一致上升,且鄂东南高于鄂西北;柑橘越冬期间低温最长持续时间每10年下降0.8 d,20世纪80年代后减少明显,但2008年出现历史次高值,属于反常年份;柑橘越冬期降水量每10年增加4.6 mm,而降水最长持续时间变化不大。49年来湖北省柑橘的综合冻害指数呈下降趋势,每10年缩短0.36;以20世纪60年代的综合冻害指数最高,至21世纪的头9年,综合冻害指数下降至最低值。在气候变暖的背景下,柑橘发生重度冻害以上的几率明显下降,但轻度、中度冻害的几率增加。

不同光照强度对广藿香中百秋李醇含量的影响

为探讨不同光照强度对广藿香[Pogostemon cablin(Blanco)Benth.]中百秋李醇含量变化的影响,采用气相色谱法对广藿香样品中的百秋李醇含量进行了测定分析。结果表明,遮阴率达80%时,广藿香中百秋李醇的含量为0.057 0%,高于全光照处理样品的0.032 5%,表明遮阴有利于广藿香中百秋李醇的积累。通过各个遮阴处理之间的多重比较,发现各遮阴处理的百秋李醇含量相互之间差异不显著(P>0.05)。因此,适当的遮阴处理对获得高品质的广藿香药材有一定的促进作用。