银杏叶提取物降低活性氧水平并激活JNK信号通路诱导人喉癌Hep-2细胞凋亡机制的研究  

The mechanism by which Ginkgo biloba extract induces the apoptosis of human laryngeal cancer Hep-2 cells by reducing reactive oxygen species level and activating JNK signaling pathway

作　　者：林海鸿 谢丹丹 胡俊[1] 潘兆虎[1] Lin Haihong;Xie Dandan;Hu Jun;Pan Zhaohu(Department of Otolaryngology,Zhejiang Provincial Taizhou Hospital,Taizhou 317000,Zhejiang Province,China)

机构地区：[1]浙江省台州医院耳鼻咽喉科,317000 [2]浙江省台州医院消化科,317000

出　　处：《中国基层医药》2021年第8期1218-1223,共6页Chinese Journal of Primary Medicine and Pharmacy

摘　　要：目的:研究银杏叶提取物(金纳多,Ginaton)对人喉癌细胞的诱导凋亡作用及其分子机制。方法:体外培养人喉癌Hep-2细胞,用金纳多以时间梯度和浓度梯度的方式处理对数生长期人喉癌Hep-2细胞,利用细胞计数试剂盒(CCK)-8实验检测金纳多对Hep-2细胞的增殖抑制作用,利用流式细胞术检测细胞凋亡情况及细胞中活性氧(ROS)水平变化,利用蛋白质免疫印迹法(Western-blot)检测凋亡和信号通路相关蛋白表达情况。结果:金纳多能够以时间依赖性和浓度依赖性的方式抑制人喉癌Hep-2细胞增殖;丙二醛(MDA)水平以时间依赖性的方式逐渐降低,处理24 h降低为2.98 μmol/g;而超氧化物歧化酶(SOD)的水平以时间依赖性的方式逐渐升高,处理24 h上升为90.35 U/g,ROS水平以时间依赖性的方式逐渐降低,处理24 h降低为18.7%,与0 h相比差异均有统计学意义( F=14.98、19.65、11.47,均 P<0.001);金纳多处理3、6、12、24 h后,磷酸化氨基末端蛋白激酶(p-JNK)的表达量依次增长为1.98、2.57、2.91、3.28,呈现出时间依赖性升高,与0 h相比差异有统计学意义( F=16.37, P<0.001)。 结论:金纳多在体外能有效抑制人喉癌Hep-2细胞增殖,诱导细胞凋亡,其作用可能与调节细胞中ROS水平并激活JNK信号通路有关。Objective To investigate the apoptosis-inducing effect of Ginkgo biloba extract(Ginaton)on human laryngeal cancer Hep-2 cells and the underlying molecular mechanism.Methods Human laryngeal cancer Hep-2 cells were cultured in vitro and human laryngeal cancer Hep-2 cells in the log phase were treated with Ginaton in time and concentration gradients.The cell counting kit-8(CCK-8)assay was performed to investigate the inhibitory effects of Ginaton on Hep-2 cells.Flow cytometry was performed to detect apoptosis and determine the level of reactive oxygen species(ROS).Western blot assay was performed to detect apoptosis and signaling pathway-related protein expression.Results Ginaton inhibited the proliferation of Hep-2 cells in a time-dependent and concentration-dependent manner.Malondialdehyde level decreased gradually in a time-dependent manner,and decreased to 2.98μmol/g after 24 hours of Ginaton treatment.Superoxide dismutase level increased gradually in a time-dependent manner and increased to 90.35 U/g after 24 hours of Ginaton treatment.ROS level decreased gradually in a time-dependent manner and deceased to 18.7%of the level before treatment after 24 hours of Ginaton treatment.There was no significant difference in ROS level between before and after 24 hours of Ginaton treatment(F=14.98,19.65,11.47,all P<0.001).After 3,6,12 and 24 hours of Ginaton treatment,the expression of phosphorylated N-terminal protein kinase increased to 1.98,2.57,2.91 and 3.28 in a time-dependent manner.There was significant difference in the expression of phosphorylated N-terminal protein kinase between before treatment and after 3,6,12 and 24 hours of Ginaton treatment(F=16.37,P<0.001).Conclusion Ginaton can effectively inhibit the proliferation and induce apoptosis of human laryngeal cancer Hep-2 cells in vitro,which may be related to regulating ROS level and activating JNK signaling pathway.

关 键 词：喉肿瘤 癌 银杏叶提取物 细胞凋亡 凋亡抑制蛋白质类 丙二醛 超氧化物歧化酶 

分 类 号：R285[医药卫生—中药学]