Anti Cancer Effects of Cnidium officinale Makino Extract Mediated through Apoptosis and Cell Cycle Arrest in the HT-29 Human Colorectal Cancer Cell Line

The anti cancer properties and underlying cell death mechanism induced by the extract of the roots of Cnidium officinale Makino(COM) were investigated. The ethanolic extract of COM inhibited the proliferation of human colon cancer cells (HT-29) in both dose-dependent and time-dependent manners. Analysis of the cell cycle after treatment of HT-29 cells with various concentrations of COM showed that COM extract inhibited the cellular proliferation of HT-29 cells via G1 phase arrest of the cell cycle. The apoptotic effect of COM on HT-29 cells was confirmed by Annexin V- propidium iodide apoptosis test. RT-PCR and Western blot analysis both revealed that COM extract dose-dependently increased the expressions of p53, p21, Bax and Caspase-3. Anti-apoptotic factor Bcl-2 expression was down regulated as well as cyclin D1 and CDK4. These data suggest that COM has anti cancer properties by inducing apoptosis and cell cycle arrest in HT-29 cells and could have possible therapeutic potential against human colon adenocarcinoma.     

蛇床子中的色原酮类化学成分及其对UMR106细胞增殖的影响

目的对蛇床Cnidium monnieri干燥成熟果实中的色原酮类化学成分进行研究,并探讨其对类成骨细胞UMR106增殖的影响。方法采用多种色谱柱技术进行分离纯化,通过波谱分析鉴定化合物结构。对所分离得到的化合物进行类成骨细胞UMR106增殖活性的测试。结果从蛇床子75%乙醇提取物中分离得到10个色原酮类化合物,分别鉴定为cnidimoside A(1)、cnidimol B(2)、前胡色原酮(3)、5,7-二羟基色原酮(4)、5-O-甲基维斯阿米醇(5)、5-O-甲基维斯阿米醇苷(6)、亥茅酚(7)、2,5-二甲基-7-羟基色原酮(8)、升麻素(9)、5-羟基色原酮-7-O-β-葡萄糖苷(10)。在样品溶液浓度为0.10 nmol/L条件下,化合物1、5、9、10对类成骨细胞UMR106的增殖促进率分别是30.23%、31.56%、35.29%和33.36%。结论化合物3~10为首次从蛇床属植物中分离得到。化合物1、5、9、10能一定程度上促进UMR106细胞的增殖。     

蛇床子超临界萃取工艺的正交设计试验研究

目的:研究蛇床子CO2超临界萃取的工艺条件。方法:以蛇床子CO2超临界萃取物中蛇床子素的含量为指标,用正交设计的方法优化选择其超临界萃取的工艺条件。结果:萃取压力、CO2流量、萃取温度三种因素对蛇床子超临界萃取物中蛇床子素的含量无显著性影响(P>0.05)。结论:萃取压力对超临界萃取的影响最大,以25mPa为宜;其次为CO2流量,以70kg/h为宜;萃取温度对超临界萃取率的影响最小。     

蛇床果实中香豆素类成分的变异及其规律

目的:探讨蛇床果实种内香豆素成分的变异与地理分布的关系及其变异规律。方法:用毛细管GC 法对蛇床果实48 个居群59 个样品中蛇床子素、佛手柑内酯、欧前胡素、O-乙酰哥伦比亚苷元等9 个香豆素成分进行含量测定与定性分析。结果与结论:根据蛇床果实中香豆素成分化学结构的差异及各成分的有无与含量的高低结合地理分布,可将蛇床分为3 个类型,即化学型I:以蛇床子素和线型呋喃香豆素为主要成分;分布于福建、浙江、江苏等亚热带常绿阔叶林区域。化学型II:以角型呋喃香豆素为主要成分;分布于辽宁、黑龙江、内蒙古等温带针阔叶混交林区域。化学型III:蛇床子素、线型和角型呋喃香豆素同时存在,属于混合的过渡类群;分布于河南、河北、山西等暖温带落叶阔叶林区域的过渡地带。     

苦参碱、氧化苦参碱与蛇床子提取物联合抗菌作用研究

目的:探讨苦参碱、氧化苦参碱和蛇床子提取物单独以及联合使用时对大肠杆菌、金黄色葡萄球菌和白色念球菌的抑制作用。方法:以苦参碱、氧化苦参碱和蛇床子提取物为研究对象,通过纸片扩散法测试其单独使用及联合使用时对上述菌种的抑菌圈直径。结果:苦参碱、氧化苦参碱单独使用时对大肠杆菌、金黄色葡萄球菌和白色念球菌均没有抑菌效果,蛇床子提取物对金黄色葡萄球菌有微弱的抑制作用,而苦参碱、氧化苦参碱与蛇床子提取物联用对金黄色葡萄球菌和白色念球菌均有很好的抑制作用。结论:本实验中苦参碱和氧化苦参碱与蛇床子提取物联合使用对测试菌种产生增效作用。     

至宝三鞭酒的薄层色谱鉴别及马钱苷的含量测定

目的:建立有效控制至宝三鞭酒质量的薄层色谱鉴别及含量测定。方法:用薄层色谱法(TLC)鉴别处方中的当归、蛇床子、何首乌、补骨脂、白芍、山茱萸、枸杞子、白术。用高效液相色谱法(HPLC)测定至宝三鞭酒中马钱苷的含量:色谱柱:Hypersi柱(250mm×4.6mm,5μm);流动相:四氢呋喃-乙晴-甲醇-0.05%磷酸溶液(1:8:4:48);检测波长:236nm;进样量:对照品进样10μL,样品进样20μL。结果:在薄层色谱上可检出当归、蛇床子2种药材的特征斑点并且无阴性干扰,可收入质量标准中。HPLC法测定中,在此色谱条件下,马钱苷在进样量为0.0185-0.7380μg范围内与峰面积呈良好的线性关系(r=0.9999,n=6)。平均加样回收率为99.6%,RSD为1.55%(n=9)。结论:TLC法鉴别至宝三鞭酒中当归、蛇床子准确,重现性好,可用于至宝三鞭酒的质量控制。HPLC法可控制至宝三鞭酒中马钱苷的含量,准确度高,重现性好,操作简便。     

川芎和引种日本川芎的RAPD分析与鉴定

目的从DNA分子水平上探讨川芎和引种日本川芎的亲缘关系及不同居群的遗传变异。方法提取35个样本的基因组DNA,并进行随机扩增多态性DNA(RAPD)分析和聚类分析。所选15个引物的RAPD,得出35个样本的聚类树状图。结果川芎和引种的日本川芎有明显的种间遗传差异,居群内遗传变异小,小生境对遗传性状有影响。结论所用方法可为川芎和日本川芎的鉴定及其道地性研究提供分子生物学依据。     

Osthole improves alcohol-induced fatty liver in mice by reduction of hepatic oxidative stress

The aim of our study was to examine the therapeutic effect of osthole, an active constituent isolated from the fruit of Cnidium monnieri (L.) Cusson, on alcohol‐induced fatty liver in mice and investigate its potential mechanisms of treatment. A mouse alcoholic fatty liver model was established by feeding 52% alcohol for 4 weeks. These experimental mice were then treated with osthole 10, 20 and 40 mg/kg for 6 weeks. The levels of serum total cholesterol (TC), triglyceride (TG), low density lipoprotein‐cholesterol (LDL‐C) and hepatic tissue contents of TC, TG and malondialdehyde (MDA) in osthole‐treated groups were significantly decreased, while the level of superoxide dismutase (SOD) was significantly increased compared with the model group. Moreover, the cytochrome P450 (CYP) 2E1 and diacylglycerol acyltransferase (DGAT) mRNA expressions in mouse liver were significantly decreased, and the carnitine palmitoyltransferase (CPT) 1A mRNA expression was increased by osthole treatment. Importantly, the histological evaluation of liver demonstrated that osthole dramatically decreased lipid accumulation. It was concluded that osthole was effective in treating mouse alcoholic fatty liver, and its main mechanisms might be related to reduction of hepatic oxidative stress, including the inhibition of reactive oxygen species (ROS) production, enhancement of antioxidative enzyme activity, and reduction of lipid accumulation and peroxidation. Copyright © 2010 John Wiley & Sons, Ltd.     

Fingerprint analysis of the fruits of Cnidium monnieri extract by high-performance liquid chromatography-diode array detection-electrospray ionization tandem mass spectrometry

A method incorporating high-performance liquid chromatography (HPLC) with electrospray ionization (ESI) and tandem mass spectrometry (MS), with parallel analysis by HPLC with UV detection using a diode-array detector (DAD) was developed for the qualitative characterization of coumarin and chromone constituents in the fruits of Cnidium monnieri. The chromatographic separations were performed on a Diamonsil C18 column (4.6 mm x 200 mm, 5 microm) with water with 50 mM ammonium acetate and 2% acetic acid (A) and acetonitrile (B) as the mobile phase. According to the characteristic UV spectra, the information of molecular weight and structure provided by ESI-MS/MS, 13 coumarin and 7 chromone components were detected and identified. This method is rapid and reliable for identification of the constituents in the complex herbal system, and the fragmentation patterns proposed could be extended to the similar compounds.     

蛇床子中喔斯脑、欧芹属素乙的提取分离及鉴定

从蛇床果实的乙醇提取物中经柱层析、薄板层析、高效液相层析证明有八种成分,已测定其中喔斯脑(Qsthol,欧芹酚—7—甲醚)及欧芹属素乙(imperatonin)为主要成分,并分离得到单体,其它六种化合物因含量少,未进一步进行分离。