中药蟾酥抗肿瘤有效/有毒物质基础及作用机制研究

项目名称： 中药蟾酥抗肿瘤有效/有毒物质基础及作用机制研究

项目编号： No.30801512

项目类型： 青年科学基金项目

立项/批准年度： 2009

项目学科： 金属学与金属工艺

项目作者： 高慧敏

作者单位： 中国中医科学院中药研究所

项目金额： 20万元

中文摘要： 蟾酥来源于蟾蜍科动物中华大蟾蜍Bufo bufo gargarizans Cantor或黑眶蟾蜍B. melanosticus Schneider耳后腺及皮肤腺的干燥分泌物。由于显著的强心作用，传统上用于心衰疾病的治疗。近年来，随着钠泵改变与肿瘤发生、发展过程相关性理论研究的深入，蟾酥及其有效成分蟾毒配基类物质的抗肿瘤作用逐渐成为关注的热点。但作为组成相对复杂、发挥多成分综合作用的中药而言，蟾酥发挥抗肿瘤作用的物质基础和作用强度尚不清晰；另外，限于狭窄的安全窗，蟾酥临床应用产生毒性的原因也有待深入研究。本课题在分析总结蟾酥及其有效成分蟾毒配基类国内外研究概况的基础上，对不同来源的蟾酥样品进行了系统的化学分离和质量分析研究；采用MTT法进行了蟾酥提取物、不同部位及单体成分的体外抗肿瘤活性筛选；针对优选出的活性部位开展了体内抗肿瘤活性评价和急性毒性研究，并采用HPLC和LC-MS/MS分析了蟾蜍总甾烯部位在小鼠和大鼠血浆及不同组织部位的代谢物。本课题的实施，有利于揭示中药蟾酥临床有效的科学内涵和发生毒副反应的作用机制，同时，为其它有毒中药的研究提供可借鉴的思路和方法。

中文关键词： 蟾酥；有毒中药；蟾毒配基；抗肿瘤；物质基础

英文摘要： Toad venom, a famous Traditional Chinese Medicine, was derived from the secretion of Bufo bufo gargarizans Cantor or B. melanostictus Schneider and widely used for the treatment of heart failure in clinic practice. Due to the finding of the function of Na+-K+-ATPase in the cancer development, the anticancer activity of toad venom and its main constituent, bufadienolides, has been increasingly paid much attention. However, As a multi-constituent TCM, toad venom included a series of compounds, such as bufadienolides and indole alkaloids. Accordingly, the effective constituents and their active mechanism of toad venom are to be investigated. Also, there is a certain risk of intoxication due to the potent cardiotonic activity and narrow therapeutic index of bufadienolides. Based on the review of toad venom and its active constituents published in the literatures, the comprehensive chemical investigation and quality evaluation of toad venom were carried out. MTT method was used for the screening of anticancer activity of toad venom extract, active fractions and pure compounds. For the screened active fraction, total bufadienolides from toad venom, its inhibitory effect against H22 and S180 tumor in mice and analgesic activity as well as acute toxicity by intra-peritoneal injection or oral administration were evaluated. The metabolites in the plasma and tissues in mice and rat were analyzed and identified by HPLC and LC-MS/MS, respectively. The present study provides the scientific evidences for making the effectiveness and toxicity mechanism of toad venom more clear. Meanwhile, the present research strategy is also feasible for the related investigation of other toxic TCMs.

英文关键词： toad venom; toxic Traditional Chinsese Medicine; bufadienolide; anticancer; active constituents