Antioxidative flavonoids from leaves of Carthamus tinctorius

A total of eight flavonoids (1-8), including a novel quercetin-7-O-(6'-O-acetyl)-beta-D-glucopyranoside (6) and seven known flavonoids, luteolin (1), quercetin (2), luteolin 7-O-beta-D-glucopyranoside (3), luteolin-7-O-(6'-O-acetyl)-beta-D-glucopyranoside (4) quercetin 7-O-beta-D-glucopyranoside (5), acacetin 7-O-beta-D-glucuronide (7) and apigenin-6-C-beta-D-glucopyrano syl-8-C-beta-D-glucopyranoside (8), have been isolated from the leaves of the safflower (Carthamus tinctorius L.) and identified on the basis of spectroscopic and chemical studies. The antioxidative activity of these flavonoids was evaluated against 2-deoxyribose degradation and rat liver microsomal lipid peroxidation induced by hydroxyl radicals generated via a Fenton-type reaction. Among these flavonoids, luteolin-acetyl-glucoside (4) and quercetin-acetyl-glucoside (6) showed potent antioxidative activities against 2-deoxyribose degradation and lipid peroxidation in rat liver microsomes. Luteolin (1), quercetin (2), and their corresponding glycosides (3 & 5) also exhibited strong antioxidative activity, while acacetin glucuronide (7) and apigenin-6,8-di-C-glucoside (8) were relatively less active.     

红花光调控信号途径关键基因CtHY5的克隆及表达分析

目的 以药用植物红花Carthamus tinctorius为研究对象,克隆光信号途径关键转录因子HY5基因,并对其进行生物信息学和表达模式分析,为红花HY5基因的功能研究提供参考。方法 以红花转录数据为参考,设计引物,采用PCR扩增方法从红花中克隆得到HY5的全长cDNA和DNA序列。运用生物信息学方法对该基因进行分析,预测编码蛋白的结构与功能,并通过荧光定量PCR方法检测CtHY5基因在红花不同组织及花发育不同时期的表达情况。结果 CtHY5基因的cDNA全长为462 bp,编码153个氨基酸,DNA全长为1941 bp,包含4个外显子和3个内含子。生物信息学分析表明,CtHY5为亲水性蛋白,定位于细胞核中。系统进化树及模体结构分析结果表明CtHY5与来自菊科的刺菜蓟、黄花蒿、薇甘菊、向日葵、莴苣中的HY5进化关系较近。定量分析表明,在白色红花和红色红花中,CtHY5基因均在花中表达量最高,其次为茎和苞片,在根中表达量最低。此外,除了根外,CtHY5基因在白色红花各组织中的表达量要明显高于红色红花。结论 首次从红花中克隆得到了光信号途径关键转录因子CtHY5基因,并研究了该基因在不同花色红花品系中的表达模式,为进一步研究其生物学功能奠定了基础。     

HPLC法测定不同来源红花中对香豆酸的含量

目的:采用高效液相色谱法测定不同来源红花中对香豆酸含量。方法:采用Diamonsil C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈-0.5%磷酸溶液(18∶82)为流动相,流速1.0 mL.min-1,检测波长308 nm,柱温30℃。结果:对香豆酸线性范围为0.0211~0.422μg.mL-1;平均回收率(n=6)为99.98%,RSD为1.2%。结论:该方法简便、准确,重复性好,可作为红花的质量控制方法。     

Systematic screening and characterization of flavonoid glycosides in Carthamus tinctorius L. by liquid chromatography/UV diode-array detection/electrospray ionization tandem mass spectrometry

The traditional Chinese medicine (TCM) is a complex system, which always consists of numerous compounds with significant difference in the content and physical and chemical properties. In this paper, a screening method based on target molecular weights was developed to characterize the flavonoid glycosides in the flower of Carthamus tinctorius L. The screening tables of aglycone and glycan were designed, respectively, in order to select and combine freely. The multiple reaction monitoring (MRM) scan mode with higher sensitivity and selectivity was adopted in the screening, which benefit the characterization for the minor components. Seventy-seven flavonoid glycosides were screened out finally, and their structures were characterized by tandem mass spectrometric method in both positive and negative ion modes. The glycosylation mode, aglycone, sequence and/or the interglycosidic linkages of the glycan portion and glycosylation position were elucidated by the fragmentation rule in the MS. Numerous compounds screened out with this method showed the structure variety in secondary plant metabolites, and the purposeful screening systemically and subsequent structure characterization offered more information about the chemical constitutions of TCM.     

不同粒度红花粉体理化性质的比较研究

 目的 比较研究不同粒度红花粉体的理化性质,确定红花超微粉体的粒度。方法 分别考察4种不同粒度红花粉体的形态结构、吸湿性、比表面积及孔容,采用HPLC测定红花粉体中羟基红花黄色素A的含量、液相图谱及体外溶出行为。结果 随红花粉体粒度的减小,其细胞破壁率增大,比表面积及孔容增加,吸湿性增加;4种粒度红花粉体中羟基红花黄色素A的含量相近,HPLC谱图相似;D₉₀为65.4与29.4 μm的红花粉体溶出度相似。结论 疏松的细胞间连接而非破壁是增加红花羟基红花黄色素A溶出的必要条件,红花超微粉体D₉₀宜控制在29.4~65.4 μm内。     

寒热性中药的成分对薄荷醇受体离子通道蛋白功能的影响

目的:探讨寒热中药成分对瞬变感受器离子通道蛋白(TRP)家族中薄荷醇受体离子通道蛋白(TRPM8)功能的影响。方法:以薄荷醇为通道激动剂,采用共聚焦显微成像法在体外观察原代培养背根神经节(DRG)神经元胞内[Ca2+]i的变化。结果:寒性中药的成分黄芩甙和大黄素均可显著上调TRPM8通道蛋白的功能;热性中药的成分桂皮醛可显著下调TR-PM8通道蛋白的功能,热性成分吴茱萸碱对TRPM8通道蛋白功能虽未见显著影响,但有下调趋势。结论:寒热性中药成分对TRPM8功能的调节可能与中药的寒热药性相关,这可能是寒热性中药临床上发挥寒热调节作用的分子机制之一。     

红花RP-HPLC指纹图谱的建立及其质量研究红花RP-HPLC指纹图谱的建立及其质量研究

目的建立红花RP-HPLC指纹图谱分析法,研究不同产地红花药材的质量。方法采用梯度洗脱的方法进行色谱分离,使用“计算机辅助相似度评价软件”进行数据处理,对不同产地红花药材指纹图谱的相似度进行比较分析。结果不同产地红花药材指纹图谱相似度较好,但仍有少数产地的药材指纹图谱中有明显差别。结论采用RP-HPLC方法控制药材的指纹图谱,方法重现性好,用于红花的质量评价切实可行。不同产地红花药材化学组成相似,其相对比例较稳定。     

红花抗心肌缺血研究进展

综述了红花抗心肌缺血的研究进展,红花制剂对治疗冠心病具明显疗效,动物整体及离体实验研究表明,红花制剂可明显改善多种动物的心肌缺血,它可减慢心率、适度降低心肌收缩力而有利于降低心肌耗氧量、改善心肌营养性供血而增加冠脉流量、抑制血小板聚集,促进血小板解聚,抑制凝血而抗血栓形成,这些作用机制可能与红花抗心肌缺血作用有关,研究红花抗心肌缺血作用机制具有重要的理论与应用前景。     

Carthamus tinctorius flower extract prevents H2O2‐induced dysfunction and oxidative damage in osteoblastic MC3T3‐E1 cells

The flowers of Carthamus tinctorius L. (Compositae) have been widely used for enhancing blood circulation and postmenopausal disorder in women. In the present study, the potential protective effects of C. tinctorius flower extract (CFE) against reactive oxygen species (ROS) induced osteoblast dysfunction were investigated using osteoblastic MC3T3‐E1 cells. The osteoblast function was assessed by measuring alkaline phosphatase activity, collagen content, calcium deposition, and RANKL production, and the oxidative status was assessed by measuring intracellular lipid peroxidation, and protein oxidation in osteoblastic MC3T3‐E1 cells. A significant reduction in the alkaline phosphatase activity, collagen, and calcium deposition and an increase in the production of receptor activator of nuclear factor‐kB ligand (RANKL) were observed after 0.3 mM H₂O₂ addition. The H₂O₂‐induced alterations were prevented by pre‐incubating the osteoblasts with 2–10 μg/ml CFE for 48 h. When the oxidative stress was induced by H₂O₂, the increased production of protein carbonyl and malondialdehyde was also reduced at the same CFE concentration. These results demonstrate that C. tinctorius flower can act as a biological antioxidant in a cell culture experimental model and protect osteoblasts from oxidative stress‐induced toxicity. Copyright © 2009 John Wiley & Sons, Ltd.     

红花化学成分研究

目的：研究红花Carthamus tinctorius的化学成分。方法：通过各种柱色谱进行分离纯化，根据理化性质和波谱数据进行结构鉴定。结果：分离并鉴定了10个化合物：7,8-dimethylpyrazino［2,3-g］quinazolin-2,4-(1H,3H)- dione（1），腺苷（2），腺嘌呤（3），尿苷（4），胸腺嘧啶（5），尿嘧啶（6），roseoside（7），4′-O-二氢红花菜豆酸-β-D-葡萄糖苷甲酯（8），4-O-β-D-吡喃葡萄糖氧基苯甲酸（9）和对羟基苯甲酸（10）。结论：化合物1和8为首次得到的天然产物，化合物7，9，10为首次从红花中分离得到。