论文部分内容阅读
目的评价来源于海洋真菌Penicillium sp.的新药候选化合物Oxalicumone A(POA)对人正常肝细胞L-02的细胞毒性作用,初步探讨POA诱导其凋亡的作用机制。方法以体外培养的L-02为实验对象,CCK-8检测细胞存活率;Hoechst 33258染色观察细胞形态学的改变;流式细胞仪检测细胞凋亡率和增殖周期;Western blot检测凋亡相关蛋白Fas、Bax以及Bcl-2的表达。结果 CCK-8检测结果表明,POA对L-02细胞有一定的时间、浓度依赖性的抑制作用;Hoechst 33258染色可见细胞出现核固缩、碎裂、浓染等典型的凋亡细胞形态学改变;流式细胞仪检测显示细胞早期凋亡率和Sub-G1、S、G2/M期出现了相应比例的增加;Western blot法表明POA显著增加了凋亡蛋白Fas、Bax表达(P<0.05),降低了抗凋亡蛋白Bcl-2表达(P<0.05)。结论 POA体外诱导L-02细胞毒活性,产生凋亡并经过线粒体信号通路,其作用机制与Bcl-2表达下调以及Fas、Bax表达上调有关。
OBJECTIVE: To evaluate the cytotoxicity of a novel drug candidate Oxalicumone A (POA) derived from marine fungus Penicillium sp. On human normal liver cell line L-02 and to explore the mechanism of POA-induced apoptosis. Methods L-02 cells were cultured in vitro, the cell viability was detected by CCK-8, the morphological changes were observed by Hoechst 33258 staining, the apoptosis rate and proliferation cycle were detected by flow cytometry, and the apoptosis-related proteins Fas, Bax and Bcl-2 expression. Results The results of CCK-8 showed that POA could inhibit L-02 cells in a time-and concentration-dependent manner. Hoechst 33258 staining showed typical morphological changes of apoptotic cells such as nuclear condensation, fragmentation and staining ; Flow cytometry showed that there was a corresponding increase in early apoptosis rate and Sub-G1, S, G2 / M phases; Western blot showed that POA significantly increased the expression of Fas and Bax (P <0.05) , Decreased the anti-apoptotic protein Bcl-2 expression (P <0.05). Conclusion POA induces the cytotoxicity of L-02 in vitro and induces apoptosis through the mitochondrial signaling pathway. The mechanism of POA is related to the down-regulation of Bcl-2 expression and the up-regulation of Fas and Bax expression.