Daphnoretin induces reactive oxygen species-mediated apoptosis in melanoma cells

Research suggests that daphnoretin exhibits a diverse array of antitumor mechanisms and pharmacological activities. However, there is no definitive explanation for the antitumor mechanisms of daphnoretin in malignant melanoma. In the present study, MTT and colony formation assays demonstrated that daphnoretin significantly inhibited the proliferation of melanoma A375 and B16 cells. Following treatment with daphnoretin, apoptotic bodies were observed in A375 and B16 cells via Hoechst 33258 staining. Furthermore, western blot analysis revealed that the apoptosis-related proteins cleaved caspase-3, cleaved caspase-9, Bax, cytochrome c and apoptotic protease-activating factor 1 were significantly upregulated, while the expression levels of caspase-3, caspase-9 and Bcl-2 were downregulated in A375 and B16 cells. Flow cytometry and fluorescence microscopy revealed that daphnoretin induced higher levels of reactive oxygen species (ROS). Therefore, the results of the present study indicated that daphnoretin induced ROS-mediated mitochondria apoptosis in human (A375) and murine (B16) malignant melanoma cells.     

Within-Plant Distribution of Coumarins in Coronilla and Securigera Species

The within-plant distribution of the coumarin compounds was investigated in Coronilla (C. vaginalis Lam., C. scorpioides (L.) Koch, C. viminalis Salisb.) and Securigera (S. varia (L.) Lassen) species with regard to improving the extraction procedure allowing better furocoumarin recovery. The hydroxycoumarins umbelliferone, scopoletin, and daphnoretin, the dihydrofurocoumarin marmesin, and the furocoumarin psoralen were detected. The hydroxycoumarins umbelliferone, scopoletin, and daphnoretin were found in all investigated species although they were not present in all the different plant organs. The dihydrofurocoumarin marmesin, never previously detected in vivo in the genus Coronilla, occurs, only in the bound form, in leaves and roots of both C. scorpioides and C. vaginalis. Also in these two species the furocoumarin psoralen was detected in all the plant parts in both free and bound forms. On the contrary, the biosynthetic pathway of furocoumarin is not operative in C. viminalis and in S. varia. The preliminary acid hydrolysis of the plant material allowed psoralen recoveries substantially higher than the simple methanolic extraction, showing that in C. scorpioides and C. vaginalis the bulk of the psoralen occurs in the bound form. With regards to the total psoralen contents, C. scorpioides appears to be a better natural source of this furocoumarin.     

Inhibition of oncogene product enzyme activity as an approach to cancer chemoprevention. Tyrosine-specific protein kinase inhibition by daphnoretin from Thymelaea hirsuta root

Inhibitors of oncogene product enzyme activity were sought as a prescreen for potential cancer chemopreventive agents. Daphnoretin, a dicoumaryl ether from Thymelaea hirsuta root, inhibited the erb-b oncogene product, the tyrosine-specific protein kinase of human epidermal growth factor receptor (IC₅₀ = 97.5 μM ). Daphnoretin was moderately cytotoxic (IC₅₀ = 21–114 μM ) only in rapidly proliferating cultured mammalian cells. © 1998 John Wiley & Sons, Ltd.     

瑞香狼毒花中香豆素成分及生物活性研究

目的 研究瑞香狼毒Stellerachamaejasme花中的香豆素类化学成分及其抗菌和抗氧化活性.方法 综合运用多种色谱分离技术进行分离纯化,并采用现代波谱学分析技术结合文献报道数据进行鉴定.对分离得到的9个香豆素类化合物进行了体内外抗氧化活性和体外抗菌活性研究.首先,采用1,1-二苯基-2-三硝基苯肼(DPPH)、...     

正交试验法优选了哥王中西瑞香素的提取工艺

目的:优选了哥王中西瑞香素的提取工艺.方法:采用单因素及正交试验,以西瑞香素含量为指标,考察超声功率、甲醇用量、提取时间、提取次数4个因素对提取工艺的影响,优选了哥王中西瑞香素的提取条件.结果:了哥王中西瑞香素的最佳提取工艺为16倍量甲醇超声提取2次,每次1.5h,超声功率160 W.结论:提取工艺简单合理可行,可用于了哥王中西瑞香素的提取.     

LC-MS/MS同时测定大鼠血浆中瑞香素、西瑞香素和瑞香新素及其药代动力学研究

建立同时测定大鼠灌胃祖师麻提取物后血浆中瑞香素、西瑞香素和瑞香新素的HPLC-MS/MS分析方法,并用于计算3个成分在大鼠体内的药代动力学参数。试验采用6只SD大鼠,口服给予祖师麻提取物(瑞香素、西瑞香素和瑞香新素的给药量分别是88.40,3.24,4.28 mg·kg~(-1)),以五味子醇甲为内标,用LC-MS/MS测定给药后血浆中药物浓度,绘制血药浓度-时间曲线,并用Kinetica 4.4软件计算相关药代动力学参数。试验结果显示,瑞香素、西瑞香素和瑞香新素的线性范围均为5~1 000μg·L~(-1),方法学考察符合要求。日内、日间变异系数(RSD)均小于15%,精密度和准确度符合生物样品分析要求。瑞香素、西瑞香素和瑞香新素口服给药后的T_(max)分别为4,2.92,2 h;C_(max)分别为858.96,178.00,36.67μg·L~(-1);AUC_(0-t)分别为10 566.4,905.89,355.11μg·L~(-1)·h;半衰期t1/2分别为5.19,3.50,4.95 h;平均驻留时间MRT分别为9.43,6.95,8.27 h。因此,该研究建立的LC-MS/MS分析方法准确灵敏,适于对SD大鼠口服祖师麻提取物后的瑞香素、西瑞香素和瑞香新素的药代动力学研究。     

了哥王的化学成分研究

目的研究了哥王Wikstroemia indica(L.)C.A.Mey.化学成分。方法运用硅胶、凝胶等多种色谱技术对了哥王的化学成分进行研究,并根据理化性质和波谱数据鉴定化合物结构。结果从了哥王85%乙醇提取物的醋酸乙酯和正丁醇萃取部位分离并鉴定了9个化合物,分别为西瑞香素(daphnoretin)、柚皮素(naringin)、3,5,7-二羟基-4’-甲氧基二氢黄酮醇(3,5,7-trihydroxy-4′-methoxy-dihydroflavonol)、对羟基苯甲酸(4-hydroxybenzoic acid)、苯甲酸(benzoic-acid)、伞形花内酯(umbdheforne)、daphnogitin、sikokianin B、芫花素(genkwanin)。结论化合物2~5为首次从该植物中分离得到。     

天山假狼毒根的化学成分研究

采用柱色谱方法对天山假狼毒Stelleropsis tianschanica根的95%乙醇提取物进行分离纯化,利用波谱技术对分得到的化合物进行结构鉴定。从天山假狼毒中分离得到了17个化合物,分别鉴定为5'-甲氧基落叶松脂醇(1)、松脂素(2)、双白瑞香素(3)、acutissimalignan B(4)、(+)-开环异落叶松树脂酚(5)、(+)-表松脂酚(6)、7-甲氧基西瑞香素(7)、thero-8S-7-methoxysyringylglycerol(8)、1-O-methyl-guaiacylglycerol(9)、2R-22'-阿魏酸酯-2,3-二羟基丙基酯(10)、vesiculosin(11)、4β,5βHguai-9,7(11)-dien-12,8-olide-1α,8α-diol(12)、(-)-nortrachelogenin(13)、4α,5βH-guai-9,7(11)-dien-12,8-olide-1α,8α-diol(14)、罗汉松脂素(15)、落叶松树脂素(16)和异落叶松树脂醇(17)。其中化合物1~13为首次从该属植物中分离得到。采用免疫荧光检测技术化合物3,7,10~14对孤儿核受体TR3的激活作用进行了考察,结果表明在50μmol·L~(-1)下,化合物10的TR3激活率高达76.38%,化合物3,7,11~14的能力较弱,均在50.0%左右。     

不同产地芫花根皮药材中西瑞香素含量比较

目的采用高效液相色谱（HPLC）法测定不同产地芫花根皮中西瑞香素含量,确定何种产地芫花根皮中西瑞香素含量最高。方法采用Prodigy 5u 100A C18柱（250 mm×4.60 mm,5μm）,流动相为甲醇-0.2%磷酸水溶液（55∶45）,流速为1.0 m L/min,柱温30℃,紫外检测波长346 nm,进样量20μL。结果西瑞香素质量浓度在0.5-20μg/m L范围内与峰面积具有良好的线性关系（r=0.998 4）,平均回收率为98.62%,RSD=4.08%（n=6）。不同产地芫花根皮药材中西瑞香素含量差异较大。结论该法可准确地对芫花根皮药材中有效成分西瑞香素进行定量定性测定,对于保证其内在质量和临床疗效具有重要的意义。     

不同剂量西瑞香素及其代谢产物在大鼠尿中的排泄

目的： 研究不同剂量西瑞香素及其葡萄糖醛酸化代谢产物在大鼠尿中的排泄动力学。 方法： 将西瑞香素制成混悬液,按50,100,200 mg·kg^-1给大鼠灌胃,于灌胃后不同时间收集尿液,用β-葡糖醛酸苷酶溶液处理尿液。采用高效液相色谱法测定尿液中西瑞香素及其葡萄糖醛酸化代谢产物的浓度。 结果： 50,100,200 mg·kg^-13种给药剂量时,累积经尿液排泄的原形药物量为（23.13±2.36）,（41.74±2.02）和（80.01±3.21）μg,累积经尿液排泄的葡萄糖醛酸结合形药物为（37.51±4.51）,（61.17±3.90）和（131.25±13.85）μg。 结论： 西瑞香素在大鼠尿液中葡萄糖醛酸化代谢产物的排泄量大于原形排泄量,原形及其葡萄糖醛酸化代谢产物的排泄呈现明显的剂量依赖性特征。