Spirostanol saponins derivated from the seeds of Trigonella foenum-graecum by β-glucosidase hydrolysis and their inhibitory effects on rat platelet aggregation

Nine spirostanol saponins (1-9) and seven mixtures of 25 R and 25 S spirostanol saponin isomers (10-16) were obtained from the seeds of Trigonella foenum-graecum after enzymatic hydrolysis of the furostanol saponin fraction by β-glucosidase. Their structures were determined by NMR and MS spectroscopy. Among them, 1- 4, 6, 8, and 9 were new compounds and five, 11B, 12A, 13B, 14A, and 14B, were new structures observed from seven mixtures. In addition, the inhibitory effects of all saponins on rat platelet aggregation were evaluated.     

Micellar aggregates of saponins from Chenopodium quinoa: characterization by dynamic light scattering and transmission electron microscopy

Entire seeds of Chenopodium quinoa Willd are a rich protein source and are also well-known for their high saponin content. Due to their amphiphily quinoa saponins are able to form intricate micellar aggregates in aqueous media. In this paper we study the aggregates formed by self-association of these compounds from two quinoa saponin fractions (FQ70 and FQ90) as well as several distinctive nanostructures obtained after their complexation with different ratios of cholesterol (CHOL) and phosphatidylcholine (PC). The FQ70 and FQ90 fractions were obtained by reversed-phase preparative chromatography. The structural features of their resulting aggregates were determined by Dynamic Light Scattering (DLS) and Transmission Electron Microscopy (TEM). Novel nanosized spherical vesicles formed by self-association with mean diameter about 100-200 nm were observed in FQ70 aqueous solutions whereas worm-like micelles an approximate width of 20 nm were detected in FQ90 aqueous solutions. Under experimental conditions similar to those reported for the preparation of Quillaja saponaria ISCOM matrices, tubular and ring-like micelles arose from FQ70:CHOL:PC and FQ90:CHOL:PC formulations, respectively. However, under these conditions no cage-like ISCOM matrices were observed. The saponin composition of FQ70 and FQ90 seems to determine the nanosized structures viewed by TEM. Phytolaccagenic acid, predominant in FQ70 and FQ90 fractions, is accountable for the formation of the nanosized vesicles and tubular structures observed by TEM in the aqueous solutions of both samples. Conversely, ring-like micelles observed in FQ90:CHOL:PC complexes can be attributed to the presence of less polar saponins present in FQ90, in particular those derived from oleanolic acid.     

Application of Electrospray Ionization Mass Spectrometry Combined with Sequential Tandem Mass Spectrometry Techniques for the Profiling of Steroidal Saponin Mixture Extracted from Tribulus terrestris

Using electrospray ionization mass spectrometry combined with sequential tandem mass spectrometry (ESI-MS (n)), the steroidal saponin mixture extracted from TRIBULUS TERRESTRIS was studied. The structures of five known steroidal saponins and two unknown steroidal saponins in the saponin mixture were investigated by sequential tandem mass spectrometry experiments. All of the steroidal saponins displayed similar fragmentation behavior in ESI-MS (n), and the characteristic cross ring cleavage pattern could be used as a fingerprint for the identification of steroidal saponins. The methodology has been established as a powerful tool for the profiling of mixtures, especially of crude plant extracts, and the structural elucidation of compounds.     

穿龙薯蓣总皂苷中甾体皂苷的分离与鉴定

目的研究穿龙薯蓣总皂苷中水溶性甾体皂苷,寻找新的生物活性化合物。方法用硅胶柱色谱、薄层色谱及高效液相色谱等进行分离,通过酸水解、理化常数和波谱学分析(IR,NMR,MS,HMQC,HMBC)鉴定化合物结构。结果从穿龙薯蓣总皂苷中分得2个甾体皂苷(1个水难溶性皂苷和1个水溶性皂苷),其化学结构分别鉴定为薯蓣皂苷元-3-O-{α-L-鼠李糖(1→2)-［β-D-葡萄糖(1→3)］}-β-D-葡糖皂苷(I),薯蓣皂苷元-3-O-［α-L-鼠李糖(1→3)-α-L-鼠李糖(1→4)-α-L-鼠李糖(1→4)］-β-D-葡糖皂苷(II)。结论II为首次从穿龙薯蓣中分离得到的新化合物,命名为穿龙薯蓣皂苷Dc。     

In vitro inhibitory effect of triterpenoidal saponins from platycodi radix on pancreatic lipase

In the process of investigating anti-obesity effect of Platycodi Radix, we found that aqueous extract of Platycodi Radix might inhibit intestinal absorption of dietary fat by inhibiting pancreatic lipase (PL) activity. In order to clarify the anti-obesity mechanism of Platycodi Radix, activity-guided isolation was performed to find active components. The total saponin fraction of Platycodi Radix appeared to have a potent inhibitory activity against the hydrolysis of triolein emulsified with phosphatidycholine by pancreatic lipase in vitro. Based on these results, further purification of active components yielded 10 known triterpenoidal saponins, among these compounds, platycodin A, C, D, and deapioplatycodin D exhibited significant inhibitory effects on PL at the concentration of 500 microg/mL with 3.3, 5.2, 34.8, and 11.67% pancreatic lipase activity vs control, respectively. Platycodin D was found to inhibit the PL activity in a dose-dependent manner. Therefore, the anti-obesity effect of Platycodi Radix might be due to the inhibition of pancreatic lipase by its saponins.     

吸收促进剂对三七总皂苷小肠吸收的影响

目的:研究三七总皂苷(panax notoginseng saponins,PNS)的小肠吸收动力学及吸收促进剂对PNS在小肠吸收速率和表观渗透系数的影响。方法:以紫外分光光度法、高效液相色谱法分别测定酚红和PNS的质量浓度,采用大鼠在体肠灌注实验考察吸收过程。结果:当药物质量浓度为40.084～400.840μg.mL-1时,大鼠小肠吸收速率常数和表观渗透系数无显著性差异(P>0.05);部分吸收促进剂能提高PNS在小肠的吸收水平,促吸收强弱顺序为:冰片>卡波姆>聚山梨酯-80,牛胆盐促吸收效果不明显,壳聚糖在一定程度上会延缓PNS的吸收。结论:PNS在大鼠小肠吸收较差,部分吸收促进剂对其吸收有一定的促进作用。     

Stimulation of LH release from cultured pituitary cells by saponins of Petersianthus macrocarpus: a permeabilizing effect.

Aqueous extracts from stem bark of Petersianthus macrocarpus contain substances exhibiting both estrogenic and antiestrogenic potency. Triterpenic saponins were identified and extracted as a bulk. Their action on the in vitro LH released by cultured rat pituitary cells was investigated. P. macrocarpus saponins stimulated the LH release in a dose-dependent manner (from 10 micrograms/ml to 300 micrograms/ml). When added simultaneously, saponins and LHRH exerted initial additive effects on LH release, demonstrating independent mechanisms of stimulation. If cells were pre-treated with saponins for 15 min, the amount of LH released under a subsequent LHRH stimulation was lowered, presumably due to a partial depletion of the cells in hormone (data not presented). However, the action of saponins on LH release did not appear specific since a general permeabilizing effect of the cell membrane was evidenced both by trypan blue exclusion and by analysis of the total protein output. When using low concentrations of saponins (10 micrograms/ml), scanning electron microscopy did not reveal any significant alteration of the cell structure, which explains why the cells remain responsive to LHRH after withdrawal of saponins. With higher concentrations (greater than 30 micrograms/ml), the same analytical studies evidenced numerous perforations of the cell membrane, with subsequent cell death. Two highly purified saponin species were tested on LH release by cultured cells; one of them (petersaponin I) appeared responsible for the observed biological effects in vitro. But as cells were shown to be efficiently protected from saponin effects by the presence of serum, it may be concluded that saponins of P. macrocarpus extracts are probably not candidate molecules promoting the in vivo estrogenic and antiestrogenic effects.     

苦瓜皂甙与黄连素混合物对Ⅱ型糖尿病小鼠的降血糖作用

目的:探讨苦瓜皂甙与黄连素混合物对实验性Ⅱ型糖尿病小鼠的降糖效果。方法:用高脂饲料喂养体重18～22g的健康昆明小鼠一周后,通过单次高剂量(150mg.kg-1)腹腔注射链脲佐菌素(STZ)的方法复制Ⅱ型糖尿病小鼠模型。将造模成功的小鼠随机分为正常空白对照组(A组)、糖尿病模型空白对照组(B组)、罗格列酮药物对照组(C组)、苦瓜皂甙组(D组),苦瓜皂甙黄连素混合物组(E组)。每天按实验设计灌胃,10天测定一次血糖,处理1个月后,分别进行糖耐量实验,观察苦瓜皂甙与黄连素混合物对Ⅱ型糖尿病小鼠血糖的影响。结果:苦瓜皂甙与黄连素混合物对Ⅱ型糖尿病小鼠有显著的降糖作用(P<0.05),在试验期内能明显改善糖耐量(P<0.05)。结论:苦瓜皂甙与黄连素混合物降糖效果明显,能明显改善糖耐量。     

Solubilizing effect and inclusion reaction of cyclic bisdesmosides from tubers of Bolbostemma paniculatum

Tubeimosides I, II and III (cyclic bisdesmosides) have been isolated from Chinese cucurbitaceous crude drug Tu-bei-mu, a tuber of Bolbostemma paniculatum (Maxim.) Franquet. Solubilizing effects of these cyclic bisdesmosides on water insoluble or less-soluble compounds were examined. It was revealed that cyclic bisdesmosides were effective on increasing the solubility of Yellow OB, dl-alpha-tocopherol and saponin A from Sapindus mukurossi. The critical micell concentration (cmc) and association number as well as diameter of micell of tubeimoside I in water were also measured. The interaction of tubeimoside I with 1-anilino-8-naphthalene-sulfonate (ANS) in aqueous solution was investigated photometrically. It was observed that tubeimoside I strongly enhanced the intensity of fluorescence of ANS, suggesting the significant formation of inclusion complex.     

Evaluation of toxicity of some saponins on brine shrimp

Toxicity of several types of saponins (1-11) against brine shrimp (Artemia salina) were evaluated. As a result, it was found that most tested compounds were not toxic to brine shrimp at high enough concentration. The most toxic saponin (1) to brine shrimp showed also cytotoxicity towards HL-60 tumor cell line using MTT assay. Brine shrimp model may thus be used as bench-top assay in finding cytotoxic components from saponin-containing fractions of plant extracts.     

Cytotoxic steroidal saponins from Agave sisalana

Two new steroidal saponins, 8 and 10, along with 7 known steroidal sapogenins and saponins (1-7) and a furostanol saponin (9) were isolated from Agave sisalana Perrine ex Engelm. The structures of these two new compounds were identified and characterized by 1D and 2D?NMR spectroscopy and mass spectrometry. In addition, acid hydrolysis and GC-FID were used to confirm the sugar moieties of 8 and 10. The cytotoxic effects of 1-10 on MCF-7, NCI-H460, and SF-268 cancer cells were evaluated, and among them, compound 10 proved to be the most cytotoxic with IC?? values of 1.2, 3.8, and 1.5 μM, respectively.     

Structure-activity relationships of saponins from Gleditsia sinensis in cytotoxicity and induction of apoptosis

Thirteen compounds were isolated from the anomalous fruits of Gleditsia sinensis on the basis of bioassay-guided fractionation. These saponins together with six analogues or related compounds were tested for their cytotoxicities against six tumor cell lines by the MTT method. The induction of apoptosis in HL-60 cells by these compounds was determined through flow cytometric analysis. Some structure-activity relationships in cytotoxicity and induction of apoptosis were identified. The evaluation of the cytotoxicity and the ability to induce apoptosis revealed that some important structural features are required for activity. A valuable model which enables prediction of their activities was established and may be employed for the drug design of new Gleditsia saponin analogues.     

Effects of quillaja saponin (Quillaja saponaria) on early embryonic zebrafish (Danio rerio) development

Although much attention has focused on environmental contamination by heavy metals, pesticides, and polychlorinated biphenyls, potential deleterious effects of naturally occurring organic compounds have received much less consideration. Saponins, which are glycosides found in many plants, are important, environmentally ubiquitous organic compounds. Saponins have both beneficial and deleterious effects in adults, but little is known about how saponins effect early vertebrate embryonic development. The authors tested the toxicity of quillaja saponin using a zebrafish embryo assay. Quillaja saponin, extracted from bark of the tree, Quillaja saponaria, is a common foaming agent used in foods and beverages. At 6 h post fertilization, zebrafish embryos were exposed to five concentrations (0 [negative control], 1, 5, 10 or 20 micro g) of quillaja saponin per milliliter of medium. Zebrafish embryos exposed to 2% ethanol were positive controls (100% embryonic death). Embryos were assessed at 30, 54, and 72 h post fertilization for changes in embryonic development, mortality, time of hatching, and morphological deformities. Embryos exposed to 1 and 5 micro g saponin were healthy, showed no obvious deformities, but exhibited shrinkage of the chorion. Hatching time for zebrafish embryos exposed to 1 and 5 micro g/ml saponin decreased by 18 h compared to unexposed embryos. Zebrafish embryos treated with 5 micro g/ml saponin responded less to touch than embryos treated with 1 micro g/ml saponin or controls. Zebrafish embryos exposed to more than 5 micro g/ml saponin exhibited 100% embryonic mortality. These results indicate that exposure to 5 micro g/ml or less of quillaja saponin acts as a growth promoter, whereas concentrations of 10 micro g/ml or greater are lethal.     

山茶种子总皂苷碱水解产物的化学成分

目的研究山茶[Camellia sinensis(L) O.Kuntze]种子总皂苷碱水解产物的化学成分。方法采用反复制备型反相HPLC法分离纯化,通过理化常数测定和波谱分析鉴定其化学结构。结果从山茶种子总皂苷的碱水解产物中分离得到5个去酰基茶皂苷类化合物,即desacyl-assamsaponin A(1)、desacyl-assamsaponin D(2)、desacyl-assamsaponin E(3)、desacyl-assamsaponin F(4)、desacyl-theasaponin E(5)。结论对山茶种子总皂苷碱水解产物化学成分系统研究尚属首次,可为山茶皂苷类化合物生物活性的构效关系研究提供参考。     

中国红参与朝鲜红参药理作用的比较——2.对糖代谢的影响

通过对不同产地的人参、不同给药剂量、给药后取材时间不同、不同动物、饱食与饥饿、去肾上腺与否等多种因素考察人参对肝糖代谢的影响。结果表明人参给药后4小时才能引起肝糖原降低,给药后8小时取材下降最为显著。引起肝糖原下降的程度高剂量比低剂量强。中国红参与朝鲜红参无论总提取物还是总皂甙均可使大鼠和小鼠的肝糖原含量下降,而中国红参提取物引起下降的幅度大于朝鲜红参。人参对肝糖代谢的途径不依赖于肾上腺,其作用强度似与糖皮质激素无关。根据机体机能状态不同(如饥饿与进食),人参对肝糖原具有双向作用(升高或降低),但是总朝着机体功能的正常化方向进行。     

Effects of saponins on the osmotic behavior of multilamellar liposomes

Effects of total ginseng saponin, 20-S-protopanaxadiol saponin, 20-S-protopanaxatriol saponin and platycodon saponin on the osmotic behavior of lipsomes were investigated by optical measurement. These saponins showed different activites on liposomal membrane, and cholesterol in liposomes was an important factor to this variation of saponin activities.     

木鳖子中总皂苷的提取分离工艺及其急性毒性的研究

目的优选木鳖子中总皂苷的最佳提取工艺,并观察木鳖子总皂苷对小鼠的急性毒性作用,评价其安全性。方法采用正交实验,以木鳖子总皂苷含量为指标,考察溶剂用量、提取次数、提取时间对木鳖子中总皂苷提取率的影响,并采用D101大孔吸附树脂结合正丁醇萃取分离提取液中木鳖子总皂苷。采用小鼠半数致死量(LD50)测定方法,观察小鼠灌胃给予木鳖子总皂苷的急性毒性反应。结果最佳提取工艺条件为:8倍量水提取三次,每次煮沸提取1 h。大孔树脂分离纯化的总皂苷纯度可达72.5%。小鼠单次灌胃给药的LD50为1.490 g.kg-1。结论优选的提取工艺结合大孔吸附树脂和正丁醇分离纯化木鳖子中总皂苷的方法,成本低且操作简便可行,适用于工业化生产木鳖子总皂苷。木鳖子总皂苷有一定的毒性。     

Modulating effects of Korean ginseng saponins on ovarian function in immature rats

The modulating effects of Korean ginseng saponins on ovarian functions were investigated in immature rats superovulated with pregnant mare serum gonadotropin (PMSG). A single dose of 1 mg (0.1 ml/head) of Korean ginseng total saponin (GTS), Korean ginseng protopanaxatriol saponin (GPT), Korean ginseng protopanaxadiol saponin (GPD), or ginsenoside-Rb1 (Gin-Rb1) was intravenously injected via jugular vein catheter three times at 1 h (early follicular phase), 25 h (middle follicular phase), and 50 h (late follicular phase) after 30 IU PMSG administration. GPD and Gin-Rb1 significantly suppressed excessive ovulatory response caused by PMSG (p<0.05). All Korean ginseng saponins significantly improved oocyte quality by decreasing the proportion of abnormal oocytes (p<0.05). Gin-Rb1 significantly decreased preovulatory serum levels of androgens and 17beta-estradiol, while GPD increased preovulatory serum progesterone level (p<0.05). GPD significantly the increased postovulatory serum progesterone level (p<0.05). These results provide strong evidence that Korean ginseng saponins have a curative effect on ovarian dysfunction caused by excessive stimulation with PMSG.     

Triterpenoid saponins from Dipsacus asper and their activities in vitro

Two new triterpenoid saponins 1 and 2, along with six known saponins 3-8, were isolated from the roots of Dipsacus asper. The structures of new compounds were established as 3-O-β-d-glucopyranosyl-(1?→?4)-[α-l-rhamnopyranosyl-(1?→?3)]-β-d-glucopyranosyl-(1?→?3)-α-l-rhamnopyranosyl-(1?→?2)-α-l-arabinopyranosyl-hederagenin-28-O-β-d-glucopyranoside (dipsacus saponin J, 1) and 3-O-α-l-arabinopyranosyl-hederagenin-28-O-β-d-glucopyranosyl-(1?→?6)-β-d-glucopyranosyl-(1?→?6)-β-d-glucopyranoside (dipsacus saponin K, 2). The structures were determined by extensive analysis of their spectroscopic data. Compounds 6 and 7 could significantly stimulate UMR106 cell proliferation and increase alkaline phosphatase activities in UMR106 cell at the concentration of 4?μM.