Total Saponins of Aralia elata （Miq.） Seem Leaves （TSAESL） Inhibits Tumor Cell Proliferation and Induces Apoptosis via activating MAPK signaling pathways in MCF-7 human breast cancer cells

Aim Aralia elata （Miq.） Seem is a medicinal plant with a variety of biological activities. Several trier- pene saponins extracted from the leaves of Aralia elata （Miq.） Seem possess potent cytotoxic activity, but the mechanisms remain unclear. Methods In the present study, we examined the anti-tumor activity of total saponins of Aralia elata （Miq.） Seem leaves （TSAESL） using both in vitro and in vivo experiments, and investigated the underlying molecular mechanisms. Results Briefly, TSAESL significantly inhibited the proliferation of five human cancer cell lines and induced apoptosis of MCF-7 cells in a concentration-dependent manner. TSAESL also in- creased the expression level of active caspase-3 and cleaved PARP. In addition, TSAESL dramatically activated the phosphorylation of ERK1/2, p38 and JNK in a concentration-dependent manner. Treatment with the ERK1/2 in- hibitor U0126, p38 inhibitor SB203580, or JNK inhibitor SP600125 prior to TSAESL exposure markedly attenuated TSAESL-induced phosphorylation of ERK1/2 and p38. In mice bearing MCF-7 xenografl, TSAESL markedly inhib- ited tumor growth without significantly affecting spleen coefficient and hematological parameters. Conclusion These results strongly suggest that TSAESL has anti-tumor effect via activating the MAPK-mediated signaling path- ways.

Comparison between Sulfuric Acid-phenol and Sulfuric Acid-anthrone Methods Used for Determination of Polysaccharides in Shoots of Aralia elata (Miq.) Seem.

Sulfuric acid-phenol and sulfuric acid-anthrone methods were used to detect polysaccharide content in shoots of Aralia elata（ Miq.） Seem.,and the conversion factor to glucose was measured with refined polysaccharides. Comprehensive evaluation was carried out by linear relationship,precision,reproducibility,stability and recovery rate. The results showed that the linear relationship between glucose concentration and absorbance was good when glucose concentration was0-40 μg/ml,and the average recovery rate was equal to or higher than 97. 00% with good reproducibility（ RSD 〈 1. 60%,n = 5）. It revealed that the two methods were accurate and reliable,and suitable for the determination of polysaccharide content in the shoots of A. elata. Polysaccharide content detected by sulfuric acid-phenol and sulfuric acid-anthrone methods was 19. 31% and 20. 40% respectively.

Geographical distribution of Aralia elata characteristics correlated with topography and forest structure in Heilongjiang and Jilin Provinces,Northeast China

The Araliaceae family consists of numerous species of medical plants of significant value as non-wood forest products.To conserve and culture these plants in natural forest stands is an important undertaking which should be implemented according to the relationship between forest structure and understory population.In this study,thirtyfive plots were established in natural A ralia elata stands.Taller individual and denser populations were found in the northern and in the eastern regions,respectively.Both population densities and individual leaf weight increased along longitude.In contrast,their relationship with elevation and DBH were negative.Along with the altitude gradient,both height and root-collar diameter increased but population density declined.Root-collar diameter and population density decreased with latitude and DBH,respectively.Overall,dominant trees unlikely enforced strong disturbance to the development of understory A.elata populations unless concerning some specific topographic factors.

Biopharmaceutical Identification of Adventitious Buds and Root Bark of Aralia elata Seem.

[Objectives]This article aimed to systematically study the biopharmaceutical characteristics of root bark and adventitious buds(tender stems and young leaves)of Aralia elata Seem.and to distinguish its root bark from that of Acanthopanax gracilistylus,in order to avoid confusion in clinical medication and guide root segment cutting technology of A.elata Seem.[Methods]Samples were obtained by field collection,and using trait and microstructure identification techniques,systematic biopharmaceutical identification of samples was carried out.[Results]The trait and microscopic characteristics of the root bark and adventitious buds(tender stems and young leaves)of A.elata Seem.were clarified.[Conclusions]The traits,cross-sections and powder of root bark,tender stems and young leaves of A.elata Seem.all have unique identification characteristics.The secondary phloem of A.elata Seem.is particularly developed,and it provides adventitious buds with sufficient nutrients for their meristem.

Aralia elata(Miq.)Seem.的木材解剖学研究

Ａｒａｌｉａ ｅｌａｔａ（ Ｍｉｑ ．）Ｓｅｅｍ 为Ａａｒｌｉａｃｅａｅ，Ａｒａｌｉａ 植物．主要分布在东北，对其进行木材解剖学研究，发现其为环孔材，射线为异形Ⅱ型。

Aralia elata inhibits neurodegeneration by downregulating O-GlcNAcylation of NF-κB in diabetic mice

AIM： To investigate the role of O-GIcNAcylation of nuclear factor-kappa B （NF-KB） in retinal ganglion cell （RGC） death and analysedthe effect of Aralia elata （AE） on neurodegen- eration in diabetic mice. METHODS： C57BL/6mice with streptozotocin-induced diabetes were fed daily with AE extract or control （CTL） diet at the onset of diabetes mellitus （DM）. Two months af- tar injection of streptozotocin or saline, the degree of cell death and the expression of O-GIcNAc transferase （OGT）, N-acetyl-b-D-glucosaminidase （OGA）, O-GIcNAcylated pro- teins, and O-GIcNAcylation of NF-KB were examined. RESULTS： AE did not affect the metabolic status of diabetic mice. The decrease in the inner retinal thickness （P〈0.001 vs CTL, P〈0.01 vs DM） and increases in RGCs with terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling （P〈0.001 vs CTL, P〈0.0001 vs DM）, glial activation, and active caspase-3 （P〈0.0001 vs CTL, P〈0.0001 vs DM） were blocked in diabetic retinas of AE extract-fed mice. Expression levels of protein O-GIcNAcylation and OGT were increased in diabetic retinas （P〈0.0001 vs CTL）, and the level of O-GIcNAcylation of the NF-KB p65 subunit was higher in diabetic retinas than in controls （P〈0.0001 vs CTL）. AE extract downregulated O-GIcNAcylation of NF-KB and prevented neurodegeneration induced by hyperglycemia （P〈0.0001 vs DM）. CONCLUSION： O-GIcNAcylation of NF-KB is concerned in neuronal degeneration and that AE prevents diabetes-in- duced RGC apoptosis via downregulation of NF-KB O-GI- cNAcylation. Hence, O-GIcNAcylation may be a new object for the treatment of DR, and AE may have therapeutic pos- sibility to prevent diabetes-induced neurodegeneration.

Three new triterpenoid saponins from Aralia echinocaulis

Objective:To study the active ingredients in the root bark of Aralia echinocaulis.Methods:Three triterpenoid saponins were separated from the 70%ethanol extracts and purified by column chromatography and their structures were determined by spectroscopic analysis.Compound 1 and 3 were evaluated for antioxidant activity by the in vitro DPPH free radical scavenging ability and the protective effect of OH-induced DNA oxidative damage.Results:Compound 1 was a new type of triterpenoid saponin,named as echinocaulisaglycone 3-O-β-Dglucopyranoside(echinocaulisaponin A),and it had good antioxidant activity.Compound 2 was similar to compound 1,named as 1-hydroxyl-echinocaulisaglycone 3-O-β-D-glucopyranoside(echinocaulisaponin B).Compound 3 was also a new type of triterpenoid saponin,named as echinocaulisaglyconeⅡ3-O-a-L-arabinopyranosyl-(1’’→4’)-β-D-glucopyranosiduronic acid(echinocaulisaponin C),and its antioxidant activity was weaker than compound 1.Conclusion:In this study,three new compounds were discovered and two of them were carried out in vitro anti-oxidation studies,laying the foundation for further research on the treatment of related diseases(cardiovascular disease,arthritis,age-related macular degeneration,etc.)through anti-oxidation or quenching free radical function.

辽东楤木叶总皂苷提取工艺研究

以总皂苷提取率、两种质量标志性皂苷成分的提取率为指标探索辽东楤木叶药材中皂苷的提取工艺。分别考察药材浸泡达饱和的时间、回流提取时乙醇溶液用量、提取时间、提取次数以及乙醇溶液浓度对辽东楤木叶皂苷提取率的影响,并采用正交实验法对辽东楤木叶总皂苷的提取工艺进行优化。结果表明,药材经乙醇水溶液浸泡3h以上达到吸收饱和;加8倍量的70%乙醇水溶液回流提取3次,每次2.5h,所得的辽东楤木叶总皂苷提取率及两种指标性皂苷提取率均较高。

热加工方式对辽东楤木营养品质及抗氧化活性的影响

采用真空蒸煮、微波、过热蒸汽3种处理方式,对辽东楤木3种抗氧化活性及总酚、总黄酮、脂肪酸、氨基酸含量进行分析。结果表明,过热蒸汽、真空蒸煮及微波处理的最佳热加工时间分别为8、20、3 min;微波加工感官评分最高;热加工处理均能提高辽东楤木抗氧化能力,过热蒸汽处理后抗氧化能力最强。与对照相比,热加工方式均显著提高辽东楤木总酚、总黄酮及总氨基酸的含量(P<0.05),其中过热蒸汽处理含量最高;微波加工的辽东楤木饱和脂肪酸含量较低,不饱和脂肪酸含量较高。总体来看,热加工处理可增加辽东楤木的营养水平;微波加工辽东楤木味道最佳,过热蒸汽加工营养品质及抗氧化能力最佳。

龙牙楤木多糖提取工艺优化及体外抗氧化活性

目的建立优化的龙牙楤木多糖水提取工艺,初步探讨其体外抗氧化活性。方法应用单因素试验结合响应面法分析料液比、提取温度、提取时间、提取次数对龙牙楤木多糖得率的影响;通过苯酚硫酸法、间羟基联苯法、考马斯亮蓝法检测龙牙楤木多糖各类成分组成;通过分析体外自由基清除能力考察龙牙楤木多糖的抗氧化活性,构建PC12细胞氧化损伤模型,考察龙牙楤木多糖体外抗氧化作用。结果按料液比m(g)V(mL)=110在91℃下水提3次,3.2 h/次,多糖得率预测值为4.38%,实际多糖得率为4.29%;响应面优化后所得多糖成分中多糖为21.39%,蛋白为7.9%,糖醛酸为1.08%;龙牙楤木多糖能够有效清除DPPH、超氧阴离子、羟基自由基,龙牙楤木多糖作用于氧化损伤的PC12细胞后,可显著提高细胞内SOD活性(P<0.05,P<0.01),降低MDA含量(P<0.05),提高细胞存活率。结论该提取方法稳定可靠,制备的龙牙楤木多糖具有明显的体外抗氧化活性,可为防治阿尔兹海默病(AD)提供一定的理论依据。

辽东楤木芽多糖乙酰化修饰工艺及其抗氧化活性研究

以辽东楤木芽为原料,采用超声波辅助提取法提取辽东楤木芽多糖,制备辽东楤木芽乙酰化多糖(Ac-AEBP)。以乙酰基取代度为指标,研究反应温度、反应时间和液料比(mL/g)对AEBP乙酰化修饰取代的影响。在单因素实验的基础上,运用Box-Behnken设计响应面实验对乙酰化工艺参数进行优化,并比较乙酰化修饰前后的辽东楤木芽多糖对·OH、·O^(2-)、DPPH·、ABTS+·的清除作用以及还原能力,探讨其抗氧化活性。结果表明辽东楤木芽多糖乙酰化最佳工艺条件为:反应温度52℃,反应时间4 h,液料比43∶1(mL/g),在此条件下,乙酰化辽东楤木芽多糖的取代度为0.4596。抗氧化研究表明,与未经修饰的AEBP相比,Ac-AEBP的抗氧化活性有较大的增强。该研究可为进一步开发利用辽东楤木芽提供科学依据。

水培茎杆规格对刺龙牙产量和营养品质的影响

为筛选出适宜刺龙牙反季节生产的水培茎杆规格,在温室内对不同长度和粗度的茎杆进行水培,研究其对顶芽萌发率、产量及营养物质(可溶性蛋白质、维生素C等)的影响。结果表明:随着水培茎杆长度和粗度的增加,顶芽萌发率和产量逐渐增加,营养成分含量逐渐提高,水培茎杆粗20 mm以上、长50 cm以上的顶芽均萌发,其中茎杆粗度达到15 mm以上、长60 cm以上时,顶芽产量均超过20 g;水培茎杆规格对顶芽多糖、可溶性糖、可溶性蛋白质和维生素C等营养成分含量的影响显著,各营养成分含量随水培茎杆粗度和长度的增加逐渐增加。结合刺龙牙整形修剪技术,建议在实际生产中水培茎杆规格以粗度(20±0.2)mm、长度50~60 cm为宜。

辽东楤木和黄花楤木的驯化栽培技术及楤木芽菜温室水培促成栽培技术

从地块选择、定植、水肥管理等方面总结辽东楤木和黄花楤木在宁夏驯化栽培技术,开展楤木嫩芽温室水培技术试验,以期为楤木人工驯化栽培和嫩芽生产提供参考。

A New Triterpenoid Saponin from Aralia subcapitata

A new oleanane-type saponin, named subcapitatoside A, has been isolated from the roots of Aralia subcapitata. On the basis of chemical and spectral evidence: the structure of subcapitatoside A has been established as: 3-O-beta-D-galactopyranosyl-(1-->2)-beta-D-galactopyranosyl oleanolic acid.

龙牙楤木总皂苷通过调控AMPK/Nrf2信号抑制缺氧/复氧诱导的心肌细胞铁死亡

目的:探究龙牙楤木总皂苷对AMPK/Nrf2信号通路的影响。方法:将人心肌细胞AC16进行体外培养,分为空白对照组、缺氧/复氧模型组、铁离子螯合剂对照组,铁离子螯合剂组、龙牙楤木总皂苷低(0.02μg/mL)浓度组、龙牙楤木总皂苷中(0.1μg/mL)浓度组、龙牙楤木总皂苷高(0.5μg/mL)浓度组,丹参滴丸(50μg/mL)阳性对照组,AMPK抑制剂Compound C(5μg/mL)+龙牙楤木总皂苷(0.5μg/mL)组,Nrf2抑制剂ML385(2μg/mL)+龙牙楤木总皂苷(0.5μg/mL)组。MTT法检测细胞存活率;试剂盒检测细胞内Fe^(2+)、ROS水平;免疫荧光法检测脂质过氧化物及GPX4、ACSL4蛋白表达;qRT-PCR检测GPX4、ACSL4 mRNA表达,Western Blot检测GPX4、ACSL4、p-AMPK、Nrf2蛋白表达;结果:与空白对照组比较,模型组细胞存活率、GPX4 mRNA表达显著降低,Fe^(2+)含量、ROS和脂质过氧化水平、ACSL4 mRNA表达显著升高,p-AMPK、Nrf2蛋白表达显著降低(P<0.01)。与模型组比较,铁离子螯合剂组细胞存活率、GPX4 mRNA表达显著升高,Fe^(2+)、ROS、脂质过氧化水平及ACSL4 mRNA表达显著降低(P<0.01);龙牙楤木总皂苷各组和丹参滴丸组细胞存活率及GPX4、p-AMPK、Nrf2蛋白表达显著升高,心肌细胞中Fe^(2+)、ROS、脂质过氧化水平及ACSL4蛋白表达显著降低(P<0.05或P<0.01)。与龙牙楤木总皂苷组比较,Compound C+龙牙楤木总皂苷组和ML385+龙牙楤木总皂苷组细胞存活率显著降低,Fe^(2+)含量显著升高,Nrf2蛋白表达显著降低,Compound C+龙牙楤木总皂苷组ROS和脂质过氧化水平显著升高,p-AMPK蛋白表达显著降低(P<0.05或P<0.01)。结论:龙牙楤木总皂苷可通过激活AMPK/Nrf2信号通路,抑制H/R诱导的心肌细胞铁死亡。

林下刺龙牙栽培技术初探

随着山野菜刺龙牙的市场需求不断增加,其野生资源面临枯竭,亟需建立林下刺龙牙栽培模式。该研究以林下人工栽植的刺龙牙为对象,探讨坡向、林分郁闭度及初植密度对刺龙牙生长的影响。结果表明:南坡栽植的刺龙牙株高、地径、顶芽基径、顶芽长度、枝梢粗度等生长指标均极显著高于北坡(P<0.01),不同坡向间成活率差异不显著(P>0.05);不同林分郁闭度下栽植的刺龙牙株高、地径、顶芽基径、顶芽长度、枝梢粗度等生长指标和成活率差异均显著(P<0.05),且郁闭度越大各指标越小;667m2栽植1400株刺龙牙的株高、地径、顶芽基径、顶芽长度、枝梢粗度等生长指标显著高于其它密度(P<0.05),各初植密度间成活率差异不显著(P>0.05)。

辽宁楤木嫩叶中三萜皂苷的分离与结构鉴定

目的:对辽宁楤木Aralia elata(Miq.)Seem.嫩叶中的三萜皂苷类成分进行分离与鉴定。方法:通过乙醇回流提取,运用硅胶、AB-8型大孔吸附树脂、ODS柱、制备HPLC进行分离纯化,利用NMR和MS谱进行化合物的结构鉴定。结果:从辽宁楤木嫩叶中分离得到15个化合物,分别鉴定为:常春藤皂苷元-3-O-β-D-吡喃葡萄糖基-(1→3)-α-L-吡喃阿拉伯糖苷(1)、楤木皂苷A(2)、刺囊酸-3-O-β-D-吡喃葡萄糖基-(1→3)-O-α-L-吡喃阿拉伯糖苷(3)、齐墩果酸-3-O-α-L-吡喃阿拉伯糖基-(1→2)-β-D-吡喃葡萄糖苷(4)、齐墩果酸-3-O-β-D-吡喃葡萄糖苷(5)、anchusosid 2(6)、齐墩果酸-3-O-β-D-吡喃葡萄糖基-(1→2)-β-D-吡喃葡萄糖苷(7)、常春藤皂苷元-3-O-β-D-吡喃葡萄糖基-(1→3)-β-D-吡喃葡萄糖苷(8)、竹节参皂苷Ⅳa(9)、randianin(10)、macranthoside A(11)、常春藤苷元-3-O-β-D-吡喃葡萄糖苷(12)、齐墩果酸-3-O-β-D-吡喃葡萄糖基-(1→2)-β-D-吡喃半乳糖苷(13)、常春藤皂苷元-3-O-α-L-呋喃阿拉伯糖基-(1→4)-β-D-吡喃葡萄糖基-28-O-β-D-吡喃葡萄糖酯苷(14)、刺囊酸-3-O-α-L-吡喃阿拉伯糖苷(15)。结论:其中,化合物4、8、11、13、14为首次从该属植物中分离得到。

龙牙楤木Aeβ-AS基因的表达对烟草中皂苷含量的影响

为探讨龙牙楤木(Aralia elata)三萜皂苷合成的关键酶基因β-香树素合成酶基因(Aeβ-AS)对三萜皂苷合成的影响。利用基因克隆技术对Aeβ-AS基因进行克隆,并对烟草进行遗传转化,分析转基因烟草中Aeβ-AS基因在各部位的表达差异,检测Aeβ-AS基因及上下游关键酶基因表达量和总三萜含量。结果表明:成功构建了植物过表达载体pROKⅡ-Aeβ-AS,并转入了野生型烟草,获得7个转基因株系,并在mRNA水平表达,且在叶子中的表达量要高于根和茎;在转基因烟草中,Aeβ-AS基因及其上下游的关键酶基因表达量均高于野生型,其中株系L21的NtFPS、NtSS基因的相对表达量最高,株系L30的NtSE、Aeβ-AS基因的相对表达量最高;与野生型烟草相比,转基因烟草的总三萜含量显著升高(1.1~1.6倍)。试验结果证明合成Aeβ-AS基因并在烟草中进行异源转化,可以使转基因烟草内的总三萜含量显著升高。

辽东楤木皂苷含量测定方法的研究

以辽东楤木为原料,探究辽东楤木总皂苷含量的测定方法,对硫酸-甲醇比色法、香草醛-浓硫酸比色法和香草醛-高氯酸比色法进行方法性能比较,实验证明硫酸-甲醇比色法操作简单,准确有效,可用于测定辽东楤木总皂苷含量,为后续辽东楤木的加工生产奠定了一定的理论基础。

基于PI3K/Akt信号通路探讨龙牙楤木总皂苷抗心肌缺血再灌注损伤的机制

目的基于磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路探讨龙牙楤木总皂苷抗心肌缺血再灌注损伤的机制。方法将60只大鼠随机分为假手术组、模型组、龙牙楤木总皂苷组、LY29004组、LY29004+龙牙楤木总皂苷组,每组10只。除假手术组外,其余组利用Langendorff离体心脏灌流装置,通过结扎左冠状动脉前降支30 min、复灌75 min的方法构建离体大鼠心脏再灌注模型。模型组采用K-H液恒速灌流;龙牙楤木总皂苷组和LY29004组分别采用含5 mg/L龙牙楤木总皂苷的K-H液和含15μmol/L LY29004的K-H液恒速灌流;LY29004+龙牙楤木总皂苷组先给予含15μmol/L LY29004的K-H液灌流15 min,再以含5 mg/L龙牙楤木总皂苷的K-H液灌流60 min。采用Western blot法检测心肌组织中PI3K/Akt信号通路相关蛋白[Akt、磷酸化蛋白激酶B(p-Akt)、糖原合酶激酶3β(GSK-3β)、磷酸化糖原合酶激酶3β(p-GSK-3β)]表达情况,免疫组化法检测心肌组织中凋亡相关蛋白[半胱氨酸天门冬氨酸酶3(Caspase-3)、B细胞淋巴瘤基因-2(Bcl-2)、Bcl-2关联蛋白X(Bax)]表达情况,HE染色观察心肌组织病理形态。结果模型组、龙牙楤木总皂苷组、LY29004组、LY29004+龙牙楤木总皂苷组Akt、p-Akt、GSK-3β、p-GSK-3β蛋白相对表达量和Caspase-3、Bax蛋白阳性表达面积均明显高于假手术组(P均<0.05),Bcl-2阳性表达面积均明显低于假手术组(P均<0.05);龙牙楤木总皂苷组Akt、p-Akt、GSK-3β、p-GSK-3β蛋白相对表达量和Bcl-2阳性表达面积均明显高于模型组、LY29004组和LY29004+龙牙楤木总皂苷组(P均<0.05),Caspase-3、Bax蛋白阳性表达面积均明显低于模型组、LY29004组和LY29004+龙牙楤木总皂苷组(P均<0.05)。假手术组心肌细胞排列有序,未见变性、坏死及炎性细胞浸润;模型组大量心肌细胞变性、坏死及炎性细胞浸润;龙牙楤木总皂苷组心肌细胞轻度肿胀,少量细胞变性、坏死,少量炎性细胞浸润;LY29004+龙牙楤木总皂苷组大量心肌细胞坏死和炎性细胞浸润。结论龙牙楤木总皂苷可能通过激活PI3K/Akt信号通路,上调抗凋亡蛋白表达和下调促凋亡蛋白表达而减轻心肌缺血再灌注损伤。