补骨脂酚通过MAPK途径诱导人肝癌HepG2细胞凋亡的研究

目的:研究补骨脂酚对人肝癌细胞凋亡的影响及其作用机制。方法:应用噻唑蓝法（MTT法）和倒置显微镜技术考察补骨脂酚对HepG2细胞生长的影响；采用荧光显微镜观察补骨脂酚处理后细胞凋亡；利用蛋白免疫印迹法检测补骨脂酚对HepG2细胞中凋亡相关蛋白及MAPK家族蛋白表达的影响；引入MAPK家族蛋白抑制剂考察生长抑制率和凋亡相关蛋白表达的变化。结果:补骨脂酚可以剂量依赖性地抑制人肝癌HepG2细胞增殖，其生长抑制作用明显强于临床上常用的抗肿瘤药5-氟尿嘧啶，并且补骨脂酚对人正常肝L02细胞具有较低的毒性。进一步研究发现，补骨脂酚可以诱导HepG2细胞发生凋亡。此外，MAPK家族参与到补骨脂酚诱导的HepG2细胞凋亡过程中，补骨脂酚可以剂量依赖性激活JNK的表达发挥促凋亡的作用，同时抑制了ERK促存活通路，然而对p38无明显影响。结论:本研究首次阐明了补骨脂酚诱导人肝癌HepG2细胞生长抑制作用机制，为进一步的临床应用提供了理论依据。

Study of bakuchiol induced apoptosis through MAPK pathways in human hepatocellular carcinoma HepG2 cells

Objective:To investigate the antitumor effect and molecular mechanism of bakuchiol against hepatocellular carcinoma cells.Methods:The growth inhibitory effect of bakuchiol on HepG2 cells was tested by MTT assay.Fluorescence microscope and inverted microscope were used to observe cell apoptosis.In addition，Western blotting detected the expression of apoptosis related proteins and MAPK family.Furthermore，the inhibitors of MAPK family protein were used to investigate the inhibitory effect and the levels of apoptosis related proteins.Results:Bakuchiol inhibited the proliferation of HepG2 cells in a dose-dependent manner，which showed stronger growth inhibitory effect than 5-fluorouracil by MTT assay.Meanwhile，bakuchiol showed low cytotoxicity in normal human liver L02 cells.Furthermore，bakuchiol could induce apoptosis in HepG2 cells.In addition，MAPK family was involved in bakuchiol-caused apoptosis.Bakuchiol dose-dependently activated the JNK MAPK，which promoted apoptosis.The activation of survival ERK was suppressed by bakuchiol，while the expression of p38 had no obvious change.Conclusion:The mechanism of bakuchiol-induced HepG2 cells growth inhibition was first elucidated in order to provide the basis for clinical application.