Cardiotonic effect of <Emphasis Type="Italic">Apocynum venetum</Emphasis> L. extracts on isolated guinea pig atrium

The effects on guinea-pig heart muscle of extracts of Apocynum venetum L. leaf, root, stem, old stem and Venetron—a polyphenol-rich extract of leaves—were studied by recording the mechanical activity and heart rate of isolated right atria. Cymarin—a cardiac glycoside—was also determined in A. venetum extracts by LC–MS/MS analysis. All extracts examined here showed a weak cardiotonic effect, i.e., induced a contractile response of the isolated atria and increased the pulse at a concentration of 1 mg/mL, which was not inhibited by propranolol (1 μM)—a β-adrenoceptor blocker. The cymarin content in extracts of A. venetum was ranked as follows: old stem >> stem > root > leaf >> Venetron. Since the cardiotonic effects of A. venetum extracts did not reflect the cymarin content, a possible mechanism other than that of cardiac glycosides was investigated. The inhibitory effects on phosphodiesterase 3 (PDE3) were studied in a cell-free enzyme assay; all extracts of various parts of A. venetum inhibited PDE purified from human platelets. These results suggest that PDE3 inhibition may contribute to the cardiotonic effects of A. venetum extracts.     

PCR-RFLP法鉴别罗布麻与混淆品白麻 及其psbA-trnH序列分析

目的:基于叶绿体psbA-trnH基因间隔区序列,建立一种PCR-RFLP(聚合酶链式反应-限制性片段长度多态性)方法鉴别罗布麻(Apocynum venetum L.)及其混淆品白麻(Apocynum pictum),并验证psbA-trnH序列酶切位点种间特异性和种内保守性.方法:将罗布麻和白麻提取总DNA,扩增psbA-trnH序列,扩增产物双向测序.序列分析发现,罗布麻含有特异性酶切位点SspⅠ.结果:扩增产物psbA-trnH为300～400 bp.罗布麻psbA-trnH产物均可以被SspⅠ酶切成两条条带,白麻却不能被SspⅠ酶切.生物信息分析结果表明,在罗布麻属中,罗布麻psbA-trnH序列的SspⅠ酶切位点具有种间特异性和种内保守性,在夹竹桃族进化树中,罗布麻与白麻距离最近.结论:本实验建立的PCR-RFLP方法可以有效鉴别罗布麻和白麻.     

Study on chemical constituents of Apocynum venetum L. leaves by LC/MS and determination of the best harvest season

The dried leaves of Apocynum venetum L. (AVL) is a traditional Chinese medicine widely used as antihypertensive medicine in China. Accurate determination of its best harvest season is important for its effective and safe use. In the present study, we developed a reliable method based on high-performance liquid chromatography-diode array detector-electro spray ionization-ion trap-time of flight (HPLC-DAD-ESI-IT-TOF) mass spectrometry and HPLC-DAD for the identification and quantification of major components in AVL leaves. Quantitative analysis of 24 samples collected weekly helped monitor the changes of compounds dynamically in AVL leaves to determine the best harvest season. A total of 30 compounds were identified, including quinic acid, five phenolic acids and 24 flavonoids. For the first time, 16 compounds were selected as marker compounds and simultaneously monitored weekly instead of monthly during the growth of the plant. The results showed that in May the leaves had the highest amount of phenolic acids, flavonoids and total compounds. Therefore, May should be the best harvest season for AVL leaves, which was distinct from previous studies. The established method was validated to be simple, accurate and precise, and thus it was of great importance for determination of the best harvest season.     

白麻叶和罗布麻叶的化学成分与抗炎活性比较研究

目的 比较研究白麻Apocynum pictum叶和罗布麻Apocynum venetum叶的化学成分及抗炎活性差异。方法 取白麻叶和罗布麻叶,20倍量60%乙醇溶液回流提取 2 次 ,每次 1.5 h ,合并提取液,浓缩、干燥,即得白麻叶提取物（APLE）、罗布麻叶提取物（AVLE）;以每毫升药液0.03 g药材的浓度过HPD300大孔树脂柱,水洗除杂后以60%乙醇溶液洗脱,收集洗脱液,浓缩、干燥,即得白麻叶精制物（APLP）、罗布麻叶精制物（AVLP）。利用UPLC/Q-TOF/MS鉴定白麻叶和罗布麻叶提取物及精制物的化学成分,进而以HPLC检测白麻苷、芦丁、异槲皮苷、金丝桃苷和紫云英苷的含量。在进行急性毒性实验确定最大耐受剂量的基础上,采用小鼠棉球肉芽肿模型考察4种样品对小鼠肉芽肿指数、抑制率和肾上腺指数的影响,并检测血清组胺、5-羟色胺和前列腺素E₂（PGE₂）水平。结果 以UPLC/Q-TOF/MS从4种样品中鉴定出化学成分97个,其中白麻叶和罗布麻叶的共有化学成分82个,主要是黄酮、黄酮糖苷和黄烷酮。HPLC检测结果显示,APLE、APLP中的白麻苷质量分数分别为 2.43%和 11.98%,罗布麻样品中未检出白麻苷,但 AVLP中芦丁、异槲皮苷、金丝桃苷和紫云英苷的质量分数分别是APLP的30.37、1.75、8.64、1.58倍。动物实验表明,与模型组比较,1.50、0.75 g·kg^-1的APLP可显著降低小鼠棉球肉芽肿指数（P＜0.05）,抑制率分别为 25.79%、21.75%;1.50 g·kg^-1剂量可显著降低血清组胺和 5-羟色胺水平（P＜0.05）,0.75 g·kg^-1剂量可显著降低 5-羟色胺水平（P＜0.01）;APLE、APLP 下调 PGE₂水平,差异不显著;AVLE、AVLP 显著升高PGE₂水平（P＜0.05）。结论 白麻叶和罗布麻叶的化学成分同中有异,相同方法制备的样品在抗炎活性上有较为明显的差别,其中APLP的抗炎活性最强,作用机制可能与调节血管活性胺的释放相关。     

罗布麻叶水提取物对链脲佐菌素致糖尿病小鼠的治疗作用及对肠道GLP-1蛋白表达的影响

目的 探讨罗布麻叶水提取物（WEAVL）对链脲佐菌素（STZ）致糖尿病模型小鼠的治疗作用。方法 随机将ICR小鼠分为对照组和WEAVL（2.34 g/kg）组,每天ig给药1次,连续30 d,检测空腹血糖和体质量。ICR小鼠按200 mg/kg ip 2% STZ溶液制备糖尿病模型,造模成功的小鼠按血糖值随机分为模型组、盐酸二甲双胍（阳性药,130 mg/kg）组和WEAVL低、中、高剂量（0.58、1.17、2.34 g/kg）组,另设对照组,连续给药30 d,测定体质量、空腹血糖、糖耐量及血糖曲线下面积（AUC）;称取肝、肾质量,计算脏器系数;取胰腺组织进行HE染色、组织病理学检查;Western blotting检测肠道组织胰高血糖素样肽-1（GLP-1）蛋白的表达水平。结果 与对照组比较,单纯给予WEAVL对正常动物空腹血糖无明显影响,可显著降低小鼠体质量（P<0.01）。与对照组比较,模型组小鼠血糖显著升高（P<0.01）,各组小鼠体质量均显著降低（P<0.01）;与模型组比较,WEAVL高剂量组给药第2~4周体质量显著降低（P<0.05、0.01）,3个剂量组2 h血糖、血糖AUC均显著降低（P<0.01）,高剂量组肝脏系数显著降低（P<0.05）、肾脏系数显著升高（P<0.05）,中、高剂量组动物胰岛细胞空泡变性例数减少,中、高剂量组肠道组织GLP-1蛋白的表达显著上调（P<0.05）。结论 WEAVL对STZ致糖尿病小鼠有一定治疗作用,其作用机制可能与上调肠道组织中GLP-1的表达有关。     

罗布麻叶水提物镇静催眠作用研究

目的 探讨罗布麻叶水提物（water extract of Apocynum venetum leaves,AVLWE）镇静催眠作用及机制。方法 随机将ICR小鼠分为对照组、右佐匹克隆（阳性药,0.40 mg·kg^-1）组和AVLWE低、中、高剂量（0.58、1.17、2.34 g·kg^-1）组,每天ig给药1次,连续4周,每周称体质量;于末次给药前1 d给药60 min后,应用旷场视频分析系统检测各组小鼠5 min自主活动;于末次给药45 min后,各组动物ip 1%戊巴比妥钠（35 mg·kg^-1,阈下催眠剂量）,记录30 min内入睡潜伏期、入睡动物数、睡眠时间;结束后麻醉处死动物,取脑称质量并计算脑系数。SD大鼠分为对照组、模型组、右佐匹克隆（阳性药,0.27 mg·kg^-1）组和AVLWE低、中、高剂量（0.40、0.81、1.62 g·kg^-1）组,每天ig给药1次,连续4周,每周称体质量;除对照组外,给药第28、29天ip对氯苯丙氨酸（PCPA）制备大鼠失眠模型,给药结束称取脑质量并计算脑系数,取下丘脑,ELISA试剂盒法测定5-羟色胺（5-HT）、多巴胺（DA）和5-羟吲哚乙酸（5-HIAA）的含量。结果 小鼠实验结果表明,与对照组比较,各给药组第1~4周体质量均显著降低（P<0.01）;右佐匹克隆片和AVLWE中、高剂量组睡眠发生率均显著增加（P<0.05、0.01）;右佐匹克隆片和AVLWE中、高剂量组睡眠时间显著延长（P<0.05）;AVLWE低、中、高剂量组脑系数均显著升高（P<0.01）。大鼠实验结果表明,与对照组比较,AVLWE高剂量组第1~4周体质量均显著降低（P<0.05、0.01）;与模型组比较,AVLWE高剂量组脑系数显著升高（P<0.05）;右佐匹克隆片组、AVLWE高剂量组DA水平显著降低（P<0.05）、5-HIAA水平显著升高（P<0.01）,AVLWE低、中、高剂量组5-HT水平显著升高（P<0.05、0.01）。结论 AVLWE具有改善睡眠的作用,机制可能与上调下丘脑5-HT水平、下调DA水平有关。     

Constituents of the leaves and roots of Ligularia stenocephala MATSUM. et KOIDZ.

Constituents of the leaves and roots of Ligularia stenocephala MATSUM. et KOIDZ. (Compositae) were investigated. Three compounds, (2E,6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyl-2,6,10,14,18,22-tetracosahexaen-1-ol, (3E,6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyl-3,6,10,14,18,22-tetracosahexaen-2-ol and neophytadiene, were isolated from the leaves of L. stenocephala. Six compounds, 5-acetyl-6-hydroxy-2α-isopropenyl-3β-methoxy-2,3H-benzofuran, 3β-acetoxy-6-acetyl-5-hydroxy-2α-isopropenyl-2,3H-benzofuran, p-hydroxybenzaldehyde, vanillin, 4-hydroxyacetophenone and 4-hydroxy-3-methoxyacetophenone, were isolated from the roots of L. stenocephala. The structures were determined on the basis of spectral data.     

金钱草中黄酮苷的分离与结构鉴定金钱草中黄酮苷的分离与结构鉴定

目的从金钱草中分离制备标准品。方法用硅胶柱色谱进行分离,通过理化方法及IR,UV,MS,¹HNMR,¹³CNMR,DEPT,HMQC,HMBC等光谱分析确定化合物结构。结果从金钱草中分离得到2个化合物,分别鉴定为鼠李酮酸γ-内酯(I)、山奈酚-3-O-α-L-鼠李糖(1→2)-β-D-木糖苷(II)。结论II为新化合物,命名为金钱草素,I为首次从该属植物中分离得到。     

Rapid identification of triterpenoid saponins in the roots of <Emphasis Type="Italic">Codonopsis lanceolata</Emphasis> by liquid chromatography–mass spectrometry

Liquid chromatography coupled with sequential mass spectrometry (LC–MS ⁿ ) has been used to identify 3,28-bidesmosidic triterpenoid saponins, lancemaside A (1), foetidissimoside A (2), aster saponin Hb (3), lancemaside E (4), lancemaside B (5), lancemaside F (6), lancemaside G (7), lancemaside C (8), and lancemaside D (9) in the roots of Codonopsis lanceolata. Structural information about both the aglycone and the sugar moiety at the C-3 position of saponins was obtained in the negative-ion mode. On the other hand, positive-ion spectra mainly provide structural information about the sugar chains of saponins, especially the oligosaccharide moiety at the C-28 position. During subsequent fragmentation of the product ions derived from the oligosaccharide moiety at the C-28 position, fragments produced by sequential loss of a monosaccharide unit were observed. Furthermore, the structural features of two unknown saponins in the roots of C. lanceolata were assigned on the basis of the fragmentation patterns of the known saponins. These studies demonstrate that LC–MS ⁿ analysis has great potential for the identification and characterization of triterpenoid saponins in plant extracts.     

Polyphenolic compounds isolated from the leaves of Myrtus communis

Four hydrolyzable tannins [oenothein B (1), eugeniflorin D₂ (2), and tellimagrandins I (3) and II (4)], two related polyphenolic compounds [gallic acid (5) and quinic acid 3,5-di-O-gallate (6)], and four myricetin glycosides [myricetins 3-O-β-d-xyloside (7), 3-O-β-d-galactoside (8), 3-O-β-d-galactoside 6″-O-gallate (9), and 3-O-α-l-rhamnoside (10)] were isolated from the leaves of Myrtus communis. Antioxidant activities of the isolated compounds were evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay.     

Simultaneous determination of seven saponins in the roots of <Emphasis Type="Italic">Codonopsis lanceolata</Emphasis> by liquid chromatography–mass spectrometry

We developed a rapid and simple analytical method for the simultaneous determination of seven 3,28-bidesmosidic triterpenoid saponins in the roots of Codonopsis lanceolata. The saponins are lancemaside A, lancemaside B, lancemaside C, lancemaside E, lancemaside G, foetidissimoside A, and aster saponin Hb. Root samples were extracted with 50% methanol and prepared for analysis. Saponins were detected by reversed-phase high-performance liquid chromatography with electrospray ionization mass spectrometry, and ginsenoside Rb₁ was used as an internal standard. The overall recoveries of all saponins were 92–116%, and the relative standard deviation values of intra- and inter-day precision were lower than 3.7 and 7.7%, respectively. Eight root samples collected from Korea and Japan were analyzed using the developed method. Lancemaside A was the most abundant saponin in the root samples from Korea, ranging from 2.65 to 3.64 mg/g dry root. However, the maximum content of lancemaside A among Japanese samples was 0.101 mg/g dry root.     

Antioxidants and inhibitor of matrix metalloproteinase-1 expression from leaves of<Emphasis Type="Italic">zostera marina</Emphasis> L.

In order to develop new anti-photoaging agents, we examined the antioxidative activity and the inhibition effect of matrix metalloproteinase-1 (MMP-1) on the extracts of a marine product, Zostera marina L., which is known for its potent activity. Three compounds (compounds 1, 2, and 3) were isolated from an ethyl acetate (EtOAc) soluble fraction of the product; they were identified as apigenin-7-O-beta-D-glucoside (1), chrysoeriol (2), and luteolin (3). These compounds were found to scavenge radicals and reactive oxygen species (ROS) and were measured to have SC50 values of 0.18 mM, 0.68 mM, and 0.01 mM against the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and 0.04 mM, 0.03 mM, and 0.01 mM against the superoxide radical in the xanthine/xanthine oxidase system, respectively. Compound 3 suppressed the expression of MMP-1 by up to 44% at 4.0 microM and inhibited the production of interleukin 6 (IL-6), which is known as a cytokine that induces MMP-1 expression. From these results, compound 3 and the other compounds were determined to have antioxidative activity and to inhibit MMP-1 expression. Thus, the three compounds are expected to be useful for preventing the photoaging of skin.     

戈宝红麻总黄酮酶法提取工艺研究

目的优化复合酶酶解辅助水提取戈宝红麻叶总黄酮的小试工艺条件。方法对提取时间、温度、料液比、酶用量、p H值等条件进行单因素考察,根据单因素考察结果设计正交试验,以总黄酮得率为指标进行考察,并进行验证试验。结果优化的提取条件:温度50℃,酶量0.16 m L·g-1,p H值为5.5,料液比为1:20,时间1.5 h,在此条件下总黄酮提取率可达到2.74%。结论酶解辅助提取戈宝红麻叶总黄酮效果较好,应用前景广阔。     

岗松中的一个新黄酮醇苷类化合物

为了研究岗松的化学成分，采用硅胶、聚酰胺柱层析色谱和重结晶的方法对化合物进行分离纯化，通过理化性质和波谱技术鉴定化合物结构。分离并鉴定了8个化合物：6，8-二甲基山柰酚-3-O-α-L-鼠李糖苷(1)、槲皮素(2)、槲皮素-3-O-α-L-鼠李糖苷(3)、杨梅素(4)、杨梅素-3-O-α-L-鼠李糖苷(5)、没食子酸(6)、熊果酸(7)和1,3-二羟基-2-(2′-甲基丙酰基)-5-甲氧基-6-甲基苯(8)。其中，化合物1为新黄酮醇苷类化合物，化合物2～7为首次从该植物中分离得到，化合物8为首次从植物中分离得到。     

Lignan glycosides from the leaves of <Emphasis Type="Italic">Osmanthus heterophyllus</Emphasis>

Seven known lignan glycosides were isolated from the leaves of Osmanthus heterophyllus: (+)-syringaresinol 4-O-β-d-glucopyranoside, (+)-syringaresinol 4, 4′-O-di-β-d-glucopyranoside, (+)-medioresinol 4, 4′-O-di-β-d-glucopyranoside, (+)-medioresinol 4-O-β-d-glucopyranoside, (+)-pinoresinol 4, 4′-O-β-d-glucopyranoside, (+)-epipinoresinol 4-O-β-d-glucopyranoside and phillyrin. Their structures were determined on the basis of spectral data.     

Two new neolignan glycosides from leaves of <Emphasis Type="Italic">Osmanthus heterophyllus</Emphasis>

Two new neolignan glycosides, (7R, 8R)-threo-guaiacylglycerol-8-O-4′-sinapyl ether 7-O-β-d-glucopyranoside (1) and (7S, 8R)-5-methoxydehydrodiconiferyl alcohol 4-O-β-d-glucopyranoside (2), and four known ones (3–6), were isolated from the leaves of Osmanthus heterophyllus. The structures of compounds 1–6 were established on the basis of spectral and chemical data.     

Two new flavonol glycosides from the leaves of Cleome chelidonii L.f.

From the leaves of Cleome chelidonii L.f., two new flavonol glycosides, named cleomesides A (1) and B (2), and four known compounds, quercetin 3-O-β-glucopyranosyl(1 → 2)-α-rhamnoside-7-O-α-rhamnoside (3), ethyl α-galactopyranoside (4), adenine (5) and glycerol monostearate (6), were isolated. The structures of all isolated compounds (1–6) were determined by NMR spectroscopy and mass spectrometry. The data of known compounds (3–6) were further compared with the reported data for these compounds.     

石松的化学成分研究

目的 研究石松全草的化学成分。方法 石松全草采用80%的乙醇回流提取,乙醇提取物经过酸溶碱沉后,用乙酸乙酯进行萃取,乙酸乙酯相经过硅胶柱层析、中压制备液相色谱及HPLC等进行分离纯化,采用现代波谱技术结合理化性质鉴定化合物结构。结果 从石松乙醇提取物中分离并鉴定了11个化合物,包括8个生物碱：α-异玉柏碱（1）、α-玉柏碱（2）、去-N-甲基-α-玉柏碱（3）、石松灵碱（4）、lycoposerramine M（5）、玉柏宁碱（6）、lyconadin D（7）、石松佛利星碱（8）;2个三萜类化合物：石松三醇（9）,α-芒柄花萜醇（10）;1个酚苷类化合物：4-glucosyloxy-3-methoxyphenl trans-propenoicethyl ester（11）。结论 化合物1为新化合物,命名为α-异玉柏碱（α-isoobscurine）。化合物7和11为首次从该植物中分离得到。     

In Vitro Studies of Intestinal Permeability and Hepatic and Intestinal Metabolism of 8-Prenylnaringenin, a Potent Phytoestrogen from Hops (Humulus lupulus L.)

Purpose The absorption potential and metabolism of 8-prenylnaringenin (8-PN) from hops (Humulus lupulus L.) were investigated. 8-PN is a potent estrogen with the potential to be used for the relief of menopausal symptoms in women. Methods Monolayers of the human intestinal epithelial cancer cell line Caco-2 and human hepatocytes were incubated with 8-PN to model its intestinal absorption and hepatic metabolism, respectively. Results The apparent permeability coefficients for 8-PN in the apical-to-basolateral and basolateral-to-apical directions of a Caco-2 monolayer were 5.2 ± 0.7 × 10⁻⁵ and 4.9 ± 0.5 × 10⁻⁵ cm/s, respectively, indicating good intestinal absorption via passive diffusion. Both glucuronide and sulfate conjugates of 8-PN were detected in the Caco-2 cell incubations. The 4′-O-glucuronide was the predominant Caco-2 cell metabolite, followed by 7-O-sulfate and 4′-O-sulfate. Both phase I and phase II metabolites of 8-PN were formed by human hepatocytes. The 7-O-glucuronide was the most abundant hepatocyte metabolite, and no sulfate conjugates were detected. Incubations with various cDNA-expressed UDP-glucuronosyltransferases indicated that the isozymes UGT1A1, UGT1A6, UGT1A8, and UGT1A9 were responsible for glucuronidation of 8-PN. Conclusions Although orally administered 8-PN should be readily absorbed from the intestine, its bioavailability should be reduced significantly by intestinal and hepatic metabolism.     

Structures of two acylated flavonol glucorhamnosides fromGinkgo biloba leaves

The position of interglycosidic linkages of two acylated flavonol glucorhamnosides fromGinkgo biloba leaves was unambiguously determined as 1→2 linkages rather than 1→4 ones on the basis of spectroscopic and chemical evidence.