| 摘要: |
| [目的] 探讨中药补骨脂活性成分补骨脂酚（BAK）诱导人肝癌Hep3B细胞生长抑制的机制。[方法] 采用噻唑蓝法（MTT法）检测BAK对人肝癌Hep3B细胞生长抑制作用；相差显微镜观察BAK处理后Hep3B细胞形态变化；Hoechst/Mitotracker/Rhodamine荧光探针染色结合高内涵细胞成像系统分析BAK对线粒体功能的影响；免疫印迹法（Western blot法）考察BAK处理后自噬标志性蛋白溶酶体相关蛋白1（LAMP1）、微管相关蛋白1轻链3（LC3）、SQSTM1蛋白（p62）以及线粒体自噬标志性蛋白PTEN诱导假定激酶1（PINK1）、帕金蛋白（Parkin）、线粒体融合蛋白2（MFN2）、线粒体膜蛋白（Tom20）的表达情况；MTT法考察线粒体自噬抑制剂氯喹（CQ）、巴弗洛霉素A1（Baf-A1）对BAK诱导的肝癌细胞生长抑制的影响。[结果] BAK时间剂量依赖性抑制Hep3B细胞生长；高内涵实验结果表明BAK能够升高线粒体质量，降低线粒体膜电位；BAK可以上调LAMP1、LC3、PINK1、Parkin表达量，下调p62、 MFN2及Tom20蛋白表达；加入线粒体自噬抑制剂可显著逆转BAK诱导的Hep3B细胞生长抑制。[结论] 补骨脂酚通过线粒体自噬抑制人肝癌Hep3B细胞生长。 |
| 关键词: 肝癌 补骨脂酚 线粒体自噬 细胞生长抑制 |
| DOI：10.11656/j.issn.1673-9043.2024.05.03 |
| 分类号:R285.5 |
| 基金项目:天津市研究生科研创新项目（2021YJSS169）；天津中医药大学研究生科研创新项目（YJSKC-20212019）；天津中医药大学中西医结合学院研究生创新基金资助项目（ZXYCXLX202013）。 |
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| Mechanism of inducing mitophagy in human hepatocellular carcinoma Hep3B cells by Bakuchiol |
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ZHAO Haonan1,2, ZHANG Qiang2, WEN Zhongqing2, KANG Ning2
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1.School of Intergrative Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China;2.School of Medical Technology, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
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| Abstract: |
| [Objective] To investigate the effect of bakuchiol（BAK） on the mitophagy of human hepatocellular carcinoma Hep3B cells and its Mechanism. [Methods] MTT assay was used to detect the effect of BAK and mitophagy inhibitors CQ/Baf-A1 on the proliferation of Hep3B cells；Morphological changes of Hep3B cells after BAK treatment observed by phase contrast microscopy；Hoechst/Mitotracker/Rhodamine fluorescent probe staining combined with high connotation cell imaging system was used to analyze the effect of BAK on mitochondrial function；The expression of autophagy-related proteins LAMP1，LC3，p62，and mitophagy-related proteins PINK1，Parkin，MFN2，Tom20 at different time points of BAK were detected by Western blotting. [Results] BAK inhibited the growth of Hep3B cells in the time- and dose-dependent manners；High Content Analysis results suggest that BAK can elevate mitochondrial mass and decrease mitochondrial membrane potential；The addition of mitophagy inhibitors markedly reverse the death rate of Hep3B cells induced by BAK；The proteins LAMP1，LC3，PINK1，Parkin were significantly increased by BAK in the time-dependent manners，while the p62，MFN2，Tom20 were reduced；Addition of mitochondrial autophagy inhibitor could significantly reverse the growth inhibition of Hep3B cells induced by BAK. [Conclusion] Bakuchiol induces inhibition of cell growth in Hep3B cells by activating mitophagy. |
| Key words: hepatocellular carcinoma bakuchiol mitophagy cell growth inhibition |
