银杏叶提取物保护顺铂致小鼠HEI-OC1细胞损伤的作用及机制研究

Effect and Mechanism of Ginkgo Biloba Extract on Cisplatin-induced HEI-OC1 Cell Injury in Mice

雷文静;陆菲;米文娟;查定军;

1:空军军医大学第一附属医院耳鼻咽喉头颈外科

摘要
目的观察银杏叶提取物对顺铂诱导小鼠耳蜗毛细胞(HEI-OC1)中AMPK及SIRT1蛋白表达水平的影响,探讨银杏叶提取物抗损伤和凋亡作用的机制。方法取对数生长期的HEI-OC1细胞,添加高糖DMEM培养基调整细胞密度到1×104/ml,将其加入6孔板中,每孔2 ml。将HEI-OC1细胞分为对照组、顺铂组、顺铂+银杏叶组,对照组HEI-OC1细胞仅采用DMEM培养基培养,顺铂组细胞中加入8μg/ml顺铂进行干预24小时,顺铂+银杏叶组预先给予HEI-OC1细胞加入50μg/ml银杏叶提取物作用24小时,再给予含有8μg/mL顺铂的DMEM培养基作用24小时;采用CCK-8试剂盒检测各组培养24、48、72和96 h时的增殖情况;采用Western blot检测三组细胞凋亡相关蛋白Caspase-3、BCL-2和Bax以及AMPK和SIRT1表达;采用流式细胞术检测三组细胞不同干预后的凋亡情况。结果 CCK-8结果显示,与对照组相比,顺铂组和顺铂+银杏叶提取物组HEI-OC1细胞的增殖能力均明显下降,在各个时间点均低于对照组(P<0.05),而顺铂+银杏叶组的HEI-OC1细胞增殖能力在48、72和96 h时均显著高于顺铂组(均为P<0.05),且在第72 h和96 h与对照组比较差异无统计学意义(P>0.05)。流式细胞术检测显示对照组细胞凋亡率为2.3%±0.2%,显著低于顺铂组(26.8%±4.3%)及顺铂+银杏叶组(7.2%±1.5%)(P<0.05)。顺铂+银杏叶组HEI-OC1细胞的Bax及Caspase-3蛋白表达水平较顺铂组显著下降,而AMPK、SIRT1及Bcl-2蛋白表达水平较顺铂组显著升高。结论银杏叶提取物能够通过AMPK/SIRT1信号通路途径抑制顺铂诱导的HEI-OC1细胞的损伤与凋亡。
关键词
银杏叶提取物;顺铂;耳蜗毛细胞;凋亡
基金项目(Foundation):
国家自然科学基金项目(81870732);; 陕西省科技重点项目(2018PT-01);陕西省科技重点项目(2017ZDXM-SF-061);; 西京助推—先进学科建设项目(XJZT14X07);; 西京助推—卫勤专项研究项目(XJZT15D02)联合资助
作者
雷文静;陆菲;米文娟;查定军;
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雷文静陆菲米文娟查定军