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  

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

作　　者：徐婵 黄慧琪 吕奕兵 林亲雄[3] 宋萍[2] 杨新洲[3] XU Chan;HUANG Huiqi;LYU Yibing;LIN Qinxiong;SONG Ping;YANG Xinzhou(Department of Pharmacy, TongjiHospital,Tongji Medical College, Huazhong University of Science and Technology,Wuhan 430030, China;Division of Science & Technology, Qinghai University for Nationalities, Xining 810007, China;School of Pharmaceutical Sciences, South-Central University for Nationalities, Wuhan 430074, China)

机构地区：[1]华中科技大学同济医学院附属同济医院药学部,武汉430030 [2]青海民族大学科学科技处,西宁810007 [3]中南民族大学药学院,武汉430074

出　　处：《华中师范大学学报（自然科学版）》2020年第5期833-840,共8页Journal of Central China Normal University：Natural Sciences

基　　金：国家自然科学基金项目(81774000);青海省重点研发与转化计划国际合作专项项目(2020-HZ-802)。

摘　　要：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±2.3μg·mL^-1、27.6±2.5μg·mL^-1,且对正常肝细胞L-02在100μg·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.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±2.3μg·mL^-1,27.6±3.2μg·mL^-1,and displayed no toxicity against normal liver cells L-02 up to 100μg·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.

关 键 词：肝癌 藏木香 抗肝癌活性 Annexin V-FITC/PI 凋亡 

分 类 号：R914.2[医药卫生—药物化学]