ERK1/2在谷氨酸诱导PC12细胞铁死亡中的作用  

Role of extracellular signal-regulated kinase 1/2 in glutamate-induced ferroptosis in PC12 cells

作　　者：黄炎 刘卓懿 张倩[1] 周婉晴 夏萍萍[1] 叶治[1] 李春玲[1] Huang Yan;Liu Zhuoyi;Zhang Qian;Zhou Wanqing;Xia Pingping;Ye Zhi;Li Chunling(Department of Anesthesiology,Xiangya Hospital,Central South University,Changsha 410008,China)

机构地区：[1]中南大学湘雅医院麻醉科,长沙410008

出　　处：《中华麻醉学杂志》2023年第8期946-950,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(82171467,82001393);湖南省自然科学基金(2021JJ3119)。

摘　　要：目的:评价细胞外调节蛋白激酶(ERK)1/2在谷氨酸诱导PC12细胞铁死亡中的作用。方法:PC12细胞采用随机数字表法分为6组(n=21):对照组(C组)、谷氨酸组(Glu组)、谷氨酸+ERK1/2过表达组(Glu+ERK1/2-OE组)、谷氨酸+ERK1/2质粒空载体组(Glu+Vec组)、谷氨酸+ERK1/2敲减组(Glu+si-ERK1/2组)和谷氨酸+ERK1/2 SiRNA阴性对照组(Glu+si-NC组)。Glu组使用终浓度为6 mmol/L的谷氨酸处理72 h,C组加入等量PBS缓冲液处理72 h,Glu+ERK1/2-OE组转染ERK1/2过表达质粒、Glu+Vec组转染质粒空载体、Glu+si-ERK1/2组转染ERK1/2 si-RNA、Glu+si-NC组转染siRNA阴性对照后48 h再使用终浓度为6 mmol/L的谷氨酸处理72 h。采用酶标仪测定细胞存活率、乳酸脱氢酶(LDH)活性、谷胱甘肽(GSH)、铁离子及丙二醛(MDA)含量,流式细胞仪测定线粒体膜电位(MMP)和脂质活性氧(Lip-ROS)水平。结果:与C组比较,Glu组细胞存活率、GSH含量和MMP下降,LDH活性、铁离子、MDA含量和Lip-ROS水平升高(P<0.05);与Glu+Vec组比较,Glu+ERK1/2-OE组细胞存活率,GSH含量及MMP升高,LDH活性、铁离子、MDA含量及Lip-ROS水平降低(P<0.05);与Glu+si-NC组比较,Glu+si-ERK1/2组细胞存活率、GSH含量及MMP降低,LDH活性、铁离子、MDA含量和Lip-ROS水平升高(P<0.05)。结论:ERK1/2参与了谷氨酸诱导PC12细胞铁死亡的过程。Objective To evaluate the role of extracellular signal-regulated kinase(ERK)1/2 in glutamate-induced ferroptosis in PC12 cells.Methods PC12 cells were divided into 6 groups(n=21 each)using a random number table method:control group(C group),glutamategroup(Glu group),glutamate+ERK1/2 over-expression group(Glu+ERK1/2-OE group),glutamate+ERK1/2 plasmid empty vector group(Glu+Vec group),glutamate+ERK1/2 knockdown group(Glu+si-ERK1/2 group)and glutamate+ERK1/2 SiRNA negative control group(Glu+si-NC group).Cells were treated with glutamate at a final concentration of 6 mmol/L for 72 h in Glu group and with the equal volume of PBS buffer for 72 h in C group.Glu+ERK1/2-OE group was transfected with ERK1/2 overexpression plasmid,Glu+Vec group was transfected with plasmid empty vector,and Glu+si-ERK1/2 group was transfected with ERK1/2 siRNA,Glu+si-NC group was transfected with siRNA negative control for 48 h,and then glutamate at a final concentration of 6 mmol/L was added and cells were treated for 72 h.The cell viability,lactic dehydrogenase(LDH)activity and contents of glutathione(GSH),ferrous ions and malondialdehyde(MDA)were measured by enzyme-linked immunosorbent assay.Mitochondrial membrane potential(MMP)and lipid reactive oxygen species(Lip-ROS)were measured by flow cytometry.Results Compared with C group,the cell viability,GSH content and MMP were significantly decreased,and the LDH activity,ferrous ions content,MDA content and Lip-ROS levels were increased in Glu group(P<0.05).Compared with Glu+Vec group,the cell viability,GSH content and MMP were significantly increased,and the activity of LDH,contents of ferrous ions and MDA,and Lip-ROS levels were decreased in Glu+ERK1/2-OE group(P<0.05).Compared with Glu+si-NC group,the cell viability,GSH content and MMP were significantly decreased,and the LDH activity,contents of ferrous ions and MDA,and Lip-ROS level were increased in Glu+si-ERK1/2 group(P<0.05).Conclusions ERK1/2 is involved in glutamate-induced ferroptosis in PC12 cells.

关 键 词：MAP激酶信号系统 谷氨酸 PC12细胞 铁死亡 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学]