枇杷叶中熊果酸的分离纯化及抗癌活性研究[C].docVIP

枇杷叶中熊果酸的分离纯化及抗癌活性研究 王涛1，邵敬伟1*，陈剑锋2，林凤屏1，郭养浩2 （1福州大学化学化工学院，福州 350108;2福州大学生物科学和工程学院，福州 350108） 摘要：目的 应用中药分离纯化技术从枇杷叶中提取纯化熊果酸，并其对人肝癌HepG2、人胃癌MGC803增殖抑制及诱导凋亡作用机理。方法 采用体外细胞培养技术，MTT法观察细胞增殖抑制作用，光镜、荧光显微镜观察细胞形态的变化，琼脂糖凝胶电泳观察DNA梯度，分光光度计检测caspase-3表达量。结果 熊果酸体外可抑制上述两株癌细胞增殖并诱导细胞凋亡，其作用机制可能与caspase-3表达量增加有关。结论 熊果酸诱导上述两株细胞凋亡，有望成为高效低毒的抗肿瘤药，具有实际开发应用价值。 关键词：熊果酸；凋亡；Caspase-3 Study on Isolation, Purification and Anti-tumor Activity of Ursolic acid from loquat leaves WANG Tao1 , SHAO Jingwei1*, CHEN Jianfeng2 , LIN Fengping1 , GUO Yanghao2 ( 1College of Chemistry and Chemical engineering , Fuzhou University , Fuzhou 350108 , China；2College of Biological Science and Technology，Fuzhou University , Fuzhou 350108, China) Abstract：Objective To purify Ursolic acid from loquat leaves,investigate anti-proliferation and apoptosis effects of Ursolic acid and explore the mechanism.Methods By cells culturing in vitro,anti-proliferation effects of Ursolic acid were assessed with MTT colorimetric assary.After treatmen with Ursolic acid,morphology of cells was observed under light microscope,fluorescence microscope respectively. agarose gel electrophoresis analysis revealed DNA cleavages,the expressions of caspase-3 were determined by the immunocytochemical method.Results Light microscope and fluorescence microscope showed that cells presented some morphologic features of apoptosis；the inhibitory effects were obvious，and the anti-proliferation of UA on hepatocarcinoma Cell HepG2、human gastric cancer cell MGC803 was in a dose-dependent and time-dependent manner; agarose gel electrophoresis analysis showed DNA cleavages（DNA ladder）;the expression of caspase-3 is increased after the cancer cell treated with UA. Conclusion UA is being expected to become high efficiency and low toxicity as a new antitumor drug. Key words: Ursolic acid;Apoptosis;Caspase-3 熊果酸(Ursolic acid，UA)，又名乌索酸，乌苏酸，属五环三萜类化合物，化学名(3β)-3-Hydroxyurs- 12-en-28-oic acid，分子式C30H48O3，分子量456.68，熔点285～291℃，易溶于二氧六环、吡啶，可溶于甲醇、乙醇、丁醇、丁酮，略溶于丙酮，微溶于苯、氯仿、