藏木香石油醚快速溶剂萃取物体外抑制肝癌细胞增殖活性研究

为充分开发利用藏药资源，对藏木香进行体外抗肝癌活性评价，并对活性部位的细胞毒活性及初步抗肝癌机制进行研究.运用快速溶剂萃取法制备藏木香的各溶剂提取物，采用MTT法、细胞形态学观察、Hoechst 33258荧光染色、流式细胞术和蛋白免疫印迹等分析藏木香各提取物抗肝癌活性及活性部位抗肝癌机制.藏木香石油醚提取物(IR-Pe)对HepG2和Hep3B都有良好的抗肝癌活性，其IC50值分别为21.9 ± 2.3 μg·mL－1、27.6 ± 2.5 μg·mL－1，且对正常肝细胞L-02在100 μg·mL－1未显示出毒性.IR-Pe对HepG2和Hep3B两个肝癌细胞株皆显示出显著的时效和量效关系；细胞形态学观察及流式细胞术显示IR-Pe明显的诱导肝癌细胞凋亡.IR-Pe明显地上调Bax和切割后的caspase-3蛋白的表达，同时下调Bcl-2蛋白的表达.IR-Pe具有良好的体外抗肝癌活性，其通过调控线粒体通路诱导肝癌细胞凋亡发挥抗肝癌活性，具有极大的开发应用潜力.

Study on inhibiting hepatocarcinoma cells proliferation of petroleum ether extract from Inula racemosa by accelerated solvent extraction in vitro

To adequately utilize and develop the Tibetan medicinal resources， we investigated the anticancer activities of extracts derived from Inula racemosa(IR)， and explored the potential mechanism.The accelerated solvent extraction method was used to prepare petroleum ether， ethyl acetate and methanolextracts from I.racemosa. MTT assay， cell morphology， Hoechst 33258 cell staining and flow cytometrywere performed to analysis the anticancer activities of the extracts and the potential mechanisms.The petroleum ether fraction from IR (IR-Pe) showed significant inhibitory activity against HepG2 and Hep3B cell lines with its IC50 values of 21.9 ± 2.3 μg·mL－1， 27.6 ± 3.2 μg·mL－1， and displayed no toxicity against normal liver cells L-02 up to 100 μg·mL－1. IR-Pe exhibited significant time-and dose-dependent cytotoxicity against HepG2 and Hep3B hepatocarcinoma cell lines. Furthermore，IR-Peobviously induced apoptosis of HepG2 and Hep3B cells by cell morphology and flow cytometry. Western blot analysis revealed that the expressions of Bax and caspase-3 increased while Bcl-2 decreased with the increase of dosage of IR-Pe. These indicated that IR-Pe suppressed hepatocellular carcinoma cells through inducing mitochondrial apoptosis and had great potential on drug development and application.