中甸前胡的化学成分研究

从伞形科植物中甸前胡(Peucedonum sp.)根中分离鉴定了1个新的香豆素化合物,命名为中旬前胡素(d-laserpitin) (1),经各项光谱测定,其结构确定为3′(S)—羟基—4′(S)—当归酰氧基—二氢邪蒿素。另外,还分离鉴定了silinidin(2),蝉翼素(Pteryxin(3),佛手柑内酯(bergapten(4),ammijin (5)等4个已知香豆素化合物和β—谷甾醇(β—sitosterol)。     

紫花前胡的提取工艺优化研究

目的:采用多指标综合评分法结合响应面法优选紫花前胡香豆素类成分的最佳提取工艺参数.方法:通过HPLC检测紫花前胡中的化学成分伞形花内酯、紫花前胡苷、补骨脂素、花椒毒素和佛手柑内酯的含量,以综合评分值作为评价指标,并对液料比、甲醇浓度和超声时间进行考察,且在单因素试验基础上,运用Box-Behnken设计响应面法优化提取...     

广西前胡的香豆素成分

从广西前胡Peucedaum guangxieuse Shan et Sheh根及根茎的乙醇提取物中,经硅胶柱层析分析8个化合物,分别鉴定为：香豆素化合物广西前胡素（peguangxenin)(1),白花前胡丁素[( )anomalin](2),伞形花内酯(umbellferone)(3),欧芹属乙素（imperatorin）(4),虎耳草素(pimpinellin)(5);其它类型化合物,falcarindiol(6),阿魏酸(ferulic acid)(7),β-谷甾醇（β-sitosterol）.其中广西前胡素（1）是新化合物,经光谱及化学方法推证其结构为3′(S)-千里光酰氧基-4′（S）-羟基-二氢邪蒿素。     

中药前胡的化学基础研究

Ⅰ.对中药前胡在植物来源、化学成分和生理活性方面的研究作了综述。Ⅱ.研究了中国西南地区应用的四种前胡品种:长前胡Peucedanum trugeniifolium)、云前胡(P. rubricaule)、中甸前胡(P.zhongdianensis)、旱前胡(Ligusticum dauoides),一种近缘植物俯卧前胡(Peucedanum decrumbens)的化学成分。从这五种药用植物中分到了33个香豆素化合物,其中有8个新香豆素,结构经光谱和化学方法证实。通过和《中华人民共和国药典》收载的法宝品种白花前胡(Peucedauum pareruporum)和紫花前胡(P. decursivum)在化学成分上的比较,我们认为,中甸前胡、长前胡和白花衣胡类似,均是比较好的前胡代用品种;旱前胡和白花前胡、紫花前胡均不类似。有待进一步的研究;草药俯卧胡香豆素含量很低,不能(也没有)作前胡应用。以上的研究为这几种药用植物的应用提供了化学成分的依据。     

俯卧前胡的化学成分

从伞形科植物俯卧前胡(Peucedanum decumbens Maxiam)的根中分离鉴定了13个成分,其中化合物(1)是一个新的二氢呋喃香豆素,经各项光谱测定,确定其结构为:顺式-2′-(1″-甲基,1″-千里光酰氧基-乙基)-3′-羟基-线型二氢呋喃香豆素,命名为俯卧前胡素(decumbensol)。     

芽前胡的化学成分

从成都产芽前胡Peucedanum turgeniifolium Wolff.中分离鉴定了12个化合物,分别为香豆素化合物佛手柑内酯(bergapten)(1),异欧芹属乙素(isoimperatorin)(2),(±)diisovaleryl-cis-khellactone(3),(±)dihydrosamidin(4),(±)peuformosin(5),(±)cis-khellactone(6),8-(2’,3’-二羟基,3’-甲基-丁基)-伞形花内酯[8-(2’3’-dihydroxy,3’-methyl-butyl)-umbelliferone](7),(±)selinidin(8),turgeniifolin A(9)以及非香豆素化合物硬脂酸(stearic acid),β-谷甾醇(β-sitosterol),甘露醇(d-mannitol)。     

伞形科中药的研究(Ⅵ) 长前胡的化学成分(第二报)

长前胡(Peucedanum turgeniifolium Wolff)全草供药用,能宣散风热,祛痰镇咳,下气;治感冒,咳嗽,痰稠,头痛及胸闷等症。为研究其药效成分,我们已从全草的甲醇抽提物中分离鉴定了五种新的二氢邪蒿素型(dihydroseselin type)二氢吡喃香豆素和甘露糖醇。其母液,我们又经硅胶柱和制备薄层层析分离,得到了另外四个香豆素化合物,经IR,MS,~1H NMR和~(13)C NMR等光谱数据解析,分别被鉴定为(±)7-羟基-8-(2′,3′-二羟基-3′-甲基-丁基)-香豆素〔(±)7-hydroxy-8-(2′,3′-dihy-droxy-3′-methyl-butyl)-coumarin〕(1),异氧化前胡素(isooxypeucedanin)(2),     

细裂前胡的香豆素成分

细裂前胡Peucedanum macilentum Franch。(伞形科)是云南西部应用的前胡地方品种,从其根的乙醇提取物中经硅胶柱层析得到6个化合物,分别鉴定为香豆素类化合物:伞形花内酯(umbelliferone)(1),佛手柑内酯(bergapten)(2),laserpitin(3),异白花前胡丁素(anomalin)(4);其它化合物:阿魏酸(ferulic acid)(5),β-谷甾醇(β-sitosterol)。     

中药前胡核磁共振氢谱法鉴定

使用＾１Ｈ－ＮＭＲ法对１９种前胡乙醚提取物进行了测试和解析,其中９种前胡的化学成分未见报道。根据主要化学成分香豆素的类型,将含角型二氢吡喃香豆素的１０种前胡归入白花前胡类;含线型二氢吡喃香豆素或线型二氢呋喃香豆素的８种前胡归秋紫花前胡类。研究结果表明,＾１Ｈ－ＮＭＲ法是鉴别中药前胡的快速、简便而可靠的检测方法。     

旱前胡的化学成分

从云南产早前胡(Ligusticum daucoides Franch.)(伞形科)根的脂溶性部分分离了11个化合物,分别为香豆素化合物:columbianadin,isoedultin,archangelicin,旱前胡甲素(daucoidin A),旱前胡乙素(dsucoidin B),佛手柑内酯(bergapten),非香豆素化合物:falcarindiol(7),阿魏酸(ferulic acid),蜡酸(cerotic acid),β-谷甾醇(β-sitosterol),胡萝卜甙(daucosterol)。其中,化合物4和5是新的香豆素化合物,经化学方法及光谱测定,证明其结构为:4是3′去乙酰基isoedultin,即:2′(S)-(1″,1″-二甲基,1″-当归酰氧基)-3′(R)-羟基-角型二氢呋喃香豆素[2′(S)-(1″,1″-dimethvl,1″-angelovloxv)—3′(R)-hydroxy angular dihydrofuranocoumarin];5是2′-(1″-甲基,1″-羟基,1″-当归酰氧亚甲基)-角型二氢呋喃香豆素[2′-(1″-methyl,1″-hydroXy,1″-angeloyloxy-methylene)angular dihydrofuranocoumarin]。     

Suppression of LPS-induced inflammatory and NF-κB responses by anomalin in RAW 264.7 macrophages

The treatment of inflammatory diseases today is largely based on interrupting the synthesis or action of the mediators that drive the host's response to injury. It is on the basis of this concept that most of the anti-inflammatory drugs have been developed. In our continuous search for novel anti-inflammatory agents from traditional medicinal plants, Saposhnikovia divaricata has been a focus of our investigations. Anomalin, a pyranocoumarin constituent of S. divaricata, exhibits potent anti-inflammatory activity. To clarify the cellular signaling mechanisms underlying the anti-inflammatory action of anomalin, we investigated the effect of anomalin on the production of inflammatory molecules in LPS-stimulated murine macrophages. The anomalin dose-dependently inhibited inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNA and protein expression in LPS-stimulated RAW 264.7 macrophage. Molecular analysis using quantitative real time polymerase chain reaction (qRT-PCR) revealed that several pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), were reduced by anomalin, and this reduction correlated with the down-regulation of the NF-κB signaling pathway. In addition, anomalin suppressed the LPS-induced phosphorylation and degradation of IκBα. To further study the mechanisms underlying its anti-inflammatory activity, an electrophoretic mobility shift assay (EMSA) using a (32) P-labeled NF-κB probe was conducted. LPS-induced NF-κB DNA binding was drastically abolished by anomalin. The present data suggest that anomalin is a major anti-inflammatory agent and may be a potential therapeutic candidate for the treatment of inflammatory disorders.     

东当归化学成分研究及定量分析

从当归属植物东当归的根的乙醇溶液中分离得到了4个化合物,通过理化特性和波谱分析分别鉴定为双(5-甲酰基糠基)醚(bis(5-formylfurfuryl)ether,1)、5-羟甲基糠醛(5-hydroxymethyl-2-furaldehyde,2)、豆甾醇(stigmasterol,3)、十七烷酸(heptadecanoic acid4,)。所有化合物均为首次从该植物中分离得到,其中1为首次从该属植物中分离得到。采用高效液相色谱法对东当归所含该属特征活性成分紫花前胡素进行了定量分析。     

Inhibitory effects of coumarin and acetylene constituents from the roots of Angelica furcijuga on D-galactosamine/lipopolysaccharide-induced liver injury in mice and on nitric oxide production in lipopolysaccharide-activated mouse peritoneal macrophages

The methanolic extract (200 mg/kg, p.o. and i.p.), principal coumarin constituents (isoepoxypteryxin, anomalin, and praeroside IV), and a polyacetylene constituent (falcarindiol) (25 mg/kg, i.p.) from the roots of Angelica furcijuga protected the liver injury induced by D-galactosamine (D-GalN)/lipopolysaccharide (LPS) in mice. In in vitro experiments, coumarin constituents (hyuganins A-D, anomalin, pteryxin, isopteryxin, and suksdorfin) and polyacetylene constituents [(-)-falcarinol and falcarindiol] substantially inhibited LPS-induced NO and/or TNF-alpha production in mouse peritoneal macrophages, and isoepoxypteryxin inhibited D-GalN-induced cytotoxicity in primary cultured rat hepatocytes. Furthermore, hyuganin A, anomalin, and isopteryxin inhibited the decrease in cell viability by TNF-alpha in L929 cells.     

Arthrinins A-D: novel diterpenoids and further constituents from the sponge derived fungus Arthrinium sp

Bioassay-guided fractionation of a methanolic extract of the fungus Arthrinium sp., isolated from the Mediterranean sponge Geodia cydonium, afforded 10 natural products including five new diterpenoids, arthrinins A-D (1-4) and myrocin D (5). In addition, five known compounds were obtained, which included myrocin A (6), norlichexanthone (7), anomalin A (8), decarboxycitrinone (9) and 2,5-dimethyl-7-hydroxychromone (10). The structures of all isolated compounds were unambiguously elucidated based on extensive 1D and 2D NMR and HR-MS analyzes. The absolute configuration of arthrinins A-D (1-4) was established by the convenient Mosher method performed in NMR tubes and by interpretation of the ROESY spectra. Antiproliferative activity of the isolated compounds was assessed in vitro against four different tumor cell lines, including mouse lymphoma (L5178Y), human chronic myelogenous leukemia (K562), human ovarian cancer (A2780) and cisplatin-resistant ovarian cancer cells (A2780CisR), using the MTT assay. Norlichexanthone (7) and anomalin A (8) exhibited the strongest activities with IC?? values ranging from 0.40 to 74.0 μM depending on the cell line investigated. This was paralleled by the inhibitory activity of both compounds against 16 cancer related protein kinases including aurora-B, PIM1, and VEGF-R2. In vitro IC?? values of 7 and 8 against these three protein kinases ranged from 0.3 to 11.7 μM. Further investigation of the potential antitumoral activity of compounds 5-8 was performed in an in vitro angiogenesis assay against human umbilical vascular endothelial cells (HUVEC) sprouting induced by vascular endothelial growth factor A (VEGF-A). Anomalin A (8), myrocin D (5) and myrocin A (6) inhibited VEGF-A dependent endothelial cell sprouting with IC?? values of 1.8, 2.6 and 3.7 μM, respectively, whereas norlichexanthone (7) was inactive.     

Decursin promotes HIF-1α proteasomal degradation and immune responses in hypoxic tumour microenvironment

BackgroundHypoxia and HIF-1α are important regulators of tumour growth and angiogenesis and could be attractive targets for cancer therapeutics. Decursin is an active compound extracted from the roots of Angelica gigas and has been shown to have potent anti-cancer and anti-angiogenic activities. However, whether decursin regulates HIF-1α activity and immune responses under hypoxic conditions is not yet understood.PurposeThe aim of this study was to identify whether decursin exhibits anti-cancer activity by targeting HIF-1α.Study designWe investigated whether decursin regulates HIF-1α protein stability and increases its degradation. In addition, we determined if decursin increases immune responses in tumour microenvironment to identify its hypoxia-associated anti-cancer activities.Materials and methodsWe performed the hypoxia-responsive element promoter–reporter assay, Western blot analysis, immune-fluorescence assay, semi-quantitative RT-PCR and ELISA for VEGF secretion, CCK-8 assay for cell proliferation, TUNEL assay for apoptosis and invasion assay in A549 human lung cancer or HCT116 human colon cancer cells. In vivo Lewis lung carcinoma (LLC) allograft mouse model was used to check tumour growth and immune responses in tumour microenvironment by immunohistochemistry analysis.ResultsWe observed that decursin inhibited HIF-1 activation under hypoxia by down-regulating the protein level of its subunit HIF-1α. It increased oxygen-dependant hydroxylation and ubiquitination of HIF-1α to promote HIF-1α degradation. Decursin also decreased mRNA expression of HIF-1α target genes. Decursin suppressed cancer cell proliferation, induced apoptosis and inhibited cancer cell invasion under hypoxia in cancer cells. In the allograft mouse tumour model, decursin reduced the hypoxic area and HIF-1α and PD-L1 expression. Infiltrating T cells (CD3+), helper T cells (CD4+) and cytotoxic (CD8+) T cells were accumulated, but regulatory T cells (Foxp3) and myeloid-derived suppressor cell-mediated immune suppressors (Arg1) were attenuated by decursin.ConclusionOur results suggest that decursin is a novel HIF-1α inhibitor that functions by promoting its proteasomal degradation and that it also helps improve T cell activation in tumour microenvironment; these findings provide new explanations about its anti-cancer and anti-angiogenic activity mechanisms.     

Decursin attenuates hepatic fibrogenesis through interrupting TGF-beta-mediated NAD(P)H oxidase activation and Smad signaling in vivo and in vitro

Aims

We studied that a potent antifibrotic effect of decursin on in vivo liver damage model and the mechanism in inhibiting which transforming growth factor (TGF)-β1-induced human hepatic stellate cells (HSCs) activation.

Main methods

Liver injury was induced in vivo by intraperitoneal injection of carbon tetrachloride (CCl₄) with or without decursin for 4 weeks in mice. Human hepatic stellate cell line, an immortalized human HSC line, was used in in vitro assay system. The effects of decursin on HSC activation were measured by analyzing the expression of α-smooth muscle actin (α-SMA) and collagen I in liver tissue and human HSCs.

Key findings

Decursin treatment significantly reduced the ratio of liver/body weight, α-SMA activation, and type I collagen overexpression in CCl₄ treated mice liver. The elevated serum levels, including ALT, AST, and ALP, were also decreased by decursin treatment. Treatment of decursin markedly proved the generation of reactive oxygen species, NAD(P)H oxidase (NOX) protein (1, 2, and 4) upregulation, NOX activity, and superoxide anion production in HSCs by TGF-β1. It also significantly reduced TGF-β1-induced Smad 2/3 phosphorylation, nuclear translocation of Smad 4, and association of Smad 2/3–Smad 4 complex. Consistent with in vitro results, decursin treatment effectively blocked the levels of NOX protein, and Smad 2/3 phosphorylation in injured mice liver.

Significance

Decursin blocked CCl₄-induced liver fibrosis and inhibited TGF-β1-mediated HSC activation in vitro. These data demonstrated that decursin exhibited hepatoprotective effects on experimental fibrosis, potentially by inhibiting the TGF-β1 induced NOX activation and Smad signaling.     

Decursin inhibits vasculogenesis in early tumor progression by suppression of endothelial progenitor cell differentiation and function

Endothelial progenitor cells (EPCs) contribute to the tumor vasculature during tumor progression. Decursin isolated from the herb Angelica gigas is known to possess potent anti‐inflammatory activities. Recently, we reported that decursin is a novel candidate for an angiogenesis inhibitor [Jung et al., 2009 ]. In this study, we investigated whether decursin regulates EPC differentiation and function to inhibit tumor vasculogenesis. We isolated AC133+ cells from human cord blood and decursin significantly decreased the number of EPC colony forming units of human cord blood‐derived AC133+ cells that produce functional EPC progenies. Decursin dose‐dependently decreased the cell number of EPC committing cells as demonstrated by EPC expansion studies. Decursin inhibited EPC differentiation from progenitor cells into spindle‐shaped EPC colonies. Additionally, decursin inhibited proliferation and migration of early EPCs isolated from mouse bone marrow. Furthermore, decursin suppressed expression of angiopoietin‐2, angiopoietin receptor Tie‐2, Flk‐1 (vascular endothelial growth factor receptor‐2), and endothelial nitric oxide synthase in mouse BM derived EPCs in a dose‐dependent manner. Decursin suppressed tube formation ability of EPCs in collaboration with HUVEC. Decursin (4 mg/kg) inhibited tumor‐induced mobilization of circulating EPCs (CD34 + /VEGFR‐2+ cells) from bone marrow and early incorporation of Dil‐Ac‐LDL‐labeled or green fluorescent protein (GFP)+ EPCs into neovessels of xenograft Lewis lung carcinoma tumors in wild‐type‐ or bone‐marrow‐transplanted mice. Accordingly, decursin attenuated EPC‐derived endothelial cells in neovessels of Lewis lung carcinoma tumor masses grown in mice. Together, decursin likely affects EPC differentiation and function, thereby inhibiting tumor vasculogenesis in early tumorigenesis. J. Cell. Biochem. 113: 1478–1487, 2012. © 2012 Wiley Periodicals, Inc.     

Inhibition of angiogenic attributes by decursin in endothelial cells and ex vivo rat aortic ring angiogenesis model

The present study was undertaken to observe the inhibition of angiogenesis by decursin. It was the first time to show that decursin offered strong anti-angiogenic activities under the biologically relevant growth (with serum) conditions. Decursin significantly inhibited human umbilical vein endothelial cell (HUVEC) proliferation concomitant with G1 phase cell cycle arrest. Decursin also inhibited HUVEC-capillary tube formation and invasion/migration in a dose-dependant manner which was associated with the suppression of matrix metalloproteinase (MMP) -2 and -9 activities. Decursin suppressed angiogenesis in ex vivo rat aortic ring angiogenesis model where it significantly inhibited blood capillary-network sprouting from rat aortic sections. Taken together, these findings suggested anti-angiogenic activity of decursin in biologically relevant condition, and warrants further pre-clinical studies for its potential clinical usefulness.     

紫伞芹的化学成分

紫伞芹(Melanosciadium pimpinelloideum de Boiss.)为伞形科紫伞芹属植物,是我国西南地区的特有种,产于四川东部及贵州西北部,生于海拔1400～1800m荫蔽潮湿的竹林或林缘草地,其化学成分未见报道。 作者从紫伞芹根部的乙醚提取物中,分离得到二个主要化学成分:川白芷素(库页当归素,anomalin)和3′当归酰基-4′-羟基-反式-凯林酮(3′-angeloyl-4′-hydroxyl-trans-khellactone)。