补骨脂酚通过激活ROS强化TRAIL的抗肝细胞癌HepG2细胞作用的机制研究  

Bakuchiol Enhanced TRAIL Induced HepG2 Cell Apoptosis through Activating Oxidative Stress

作　　者：肖嘎 杨锐 吴涛[3] 党玲 宋涛[1] 董昌利 XIAO Ga;YANG Ru;WU Tao;DANG Ling;SONG Tao;DONG Chang-li(Department of General Surgery,Second Affliated Hospital of Xi＇an Medical College,Xi＇an,Shaanxi,710038,China;Internal Medicine Departmeng Gucheng People＇s Hospital,Hengshui,Hebei,253800,China;Department of General Surgery,Tangdu Hospital,Fourth military medical university,Xi＇un,Shaanxi,710038,China)

机构地区：[1]西安医学院第二附属医院普外科,陕西西安710038 [2]故城县医院内五科,河北衡水253800 [3]第四军医大学附属唐都医院普外科,陕西西安710038

出　　处：《现代生物医学进展》2018年第20期3835-3839,3853,共6页Progress in Modern Biomedicine

基　　金：陕西省科学技术研究发展计划项目(2013K12-03-06)

摘　　要：目的:探究补骨脂酚(Bakuchiol,Bak)对肿瘤坏死因子相关凋亡诱导配体(Tumor necrosis factor-related apoptosis-inducing ligand,TRAIL)抗HepG2细胞作用的影响及内在机制。方法:常规培养HepG2细胞,给予梯度浓度的Bak处理,检测细胞活力。联合应用Bak与TRAIL处理,检测细胞活力。Western blot检测Bak处理后氧化应激水平、死亡受体4(Death Receptor 4,DR4)、DR5的表达变化。联合应用Bak与TRAIL检测凋亡情况。进而引入ROS清除剂NAC,联合NAC处理后,检测ROS、DR4、DR5以及凋亡情况。结果:Bak剂量依赖地抑制了HepG2细胞的活力,联合应用Bak+TRAIL对细胞活力的抑制作用优于单独用药。Bak处理后氧化应激水平升高,体现在ROS增加,GSH水平下降;Western blot检测发现Bak处理后DR4、DR5表达增加。联合应用Bak+TRAIL显著增加了细胞凋亡蛋白Bax的表达,抑制了抗凋亡蛋白Bcl2的表达。引入ROS阻断剂NAC处理后,与Bak+TRAIL组相比,ROS水平下降,DR4、DR5表达减少。凋亡检测发现NAC处理降低了Bak+TRAIL引起的细胞凋亡。结论:Bak可以显著增强TRAIL引起的HepG2细胞凋亡,该作用可能与Bak激活氧化应激进而上调DR4、DR5表达有关。Objective： To investigate the roles of Bakuchiol（Bak） on HepG2 cell and the effects of co-treatment of Bak and Tumor necrosis factor-related apoptosis-inducing ligand（TRAIL） on HepG2 cells, as well as the inner mechanisms. Methods： Cells were treated with different concentrations of Bak or Bak＋TRAILand cell vitality was detected 24 h later. After Bak treatment, cell oxidative stress levels were examined through DCF-DA staining and ROS, GSH detection. Western blot analysis was conducted to detected the expressions of Death receptor 4（DR4） and DR5. After Bak＋TRAIL treatment, cell apoptosis status was detected. Furthermore, ROS scavenger NAC was used to inhibit oxidative stress and DR4, DR5, Bax expressions were detected. Results： Both Bak and TRAIL can reduce HepG2 cell vitality. Co-treatment of Bak and TRAIL further reduced the cell vitality. Bak treatment increased ROS production,DR expressions and reduced cellular GSH level. Co-treatment of Bak and TRAIL induced higher level of apoptosis compared with single-drug treatment. Compared with the Bak ＋ TRAIL group, NAC treatment reduced ROS level and reduced DR expression, as well as the Bak＋TRAIL induced apoptosis levels. Conclusion： Bak enhances TRIAL induced HepG2 cell apoptosis and tumor growth via activating cellular oxidative stress and up regulating DR4 and DR5 expressions.

关 键 词：肝细胞肝癌 补骨脂酚 肿瘤坏死因子相关凋亡诱导配体 氧化应激 死亡受体 

分 类 号：R-33[医药卫生] F735.7[经济管理—产业经济]