不全羧化骨钙素对HT22海马神经元铁死亡的抑制作用  

Inhibitory effect of undercarboxylatedosteocalcin on ferroptosis in HT22 hippocampal neurons

作　　者：吴嘉美 郑静 岳向培 罗常溢 刘水蓉 王妍 赵云[1] WU Jiamei;ZHENG Jing;YUE Xiangpei;LUO Changyi;LIU Shuirong;WANG Yan;ZHAO Yun(Institute of Military Cognition and Brain Sciences,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;Anhui Medical University,Hefei 230032,China)

机构地区：[1]军事科学院军事医学研究院军事认知与脑科学研究所,北京100850 [2]安徽医科大学,合肥230032

出　　处：《军事医学》2023年第10期740-747,共8页Military Medical Sciences

基　　金：国家重点研发计划(2016YFE0204400)。

摘　　要：目的探讨不全羧化骨钙素(ucOCN)对Erastin诱导小鼠海马神经元HT22铁死亡的作用。方法以HT22海马神经元为研究对象,采用Erastin处理HT22细胞诱导铁死亡,实验分为对照组、ucOCN组、Erastin组、Erastin+ucOCN组。采用CCK⁃8试剂盒检测各组细胞存活率,光学显微镜下观察细胞形态变化,Live/Dead染色检测存活及死亡细胞数目,超氧化物阴离子荧光探针(DHE)检测细胞活性氧(ROS)含量,TMRE探针检测线粒体膜电位,实时荧光定量PCR(qRT⁃PCR)验证siRNA对OCN的干扰效率,Western印迹实验检测细胞铁死亡相关蛋白谷胱甘肽过氧化物酶4(GPX4)和溶质载体家族7成员11(SLC7A11)蛋白的表达水平。结果30µmol/L Erastin作用于HT22细胞24 h明显降低细胞存活率,诱导细胞死亡,增加细胞内ROS含量,并降低线粒体膜电位。与Erastin组相比,Erastin+ucOCN组细胞存活率有所升高,细胞内ROS水平显著降低,线粒体膜电位有所恢复;Western印迹实验结果显示,ucOCN能够促进铁死亡细胞内GPX4和SLC7A11蛋白的表达。结论ucOCN对Erastin诱导细胞铁死亡具有抑制作用。Objective To investigate the effect of undercarboxylatedosteocalcin(ucOCN)on erastin⁃induced ferroptosis in mouse hippocampal HT22 neurons.Methods HT22 hippocampal neurons were treated with erastin to induce a ferroptosis model.The cells were divided into the control group,ucOCN group,erastin group,and erastin+ucOCN group.CCK⁃8 analysis was used to detect the proliferation of cells in each group.The morphological changes of cells in different groups were observed under optical microscope,and the number of surviving and dead cells were detected by Live/Dead staining.Superoxide anion fluorescent probe(DHE)was used to detect the content of reactive oxygen species(ROS),and TMRE probe was used to detect the mitochondrial membrane potential.Quantitative real⁃time polymerase chain reaction(qRT⁃PCR)was verified the interference efficiency of siRNA on OCN.The protein expression levels of ferroptosis⁃related proteins⁃glutathione peroxidase 4(GPX4)and solute carrier family 7,member 11(SLC7A11)were detected by Western blotting.Results Twenty⁃four hours of treatment of HT22 cells with 30µmol/L erastin significantly inhibited cell viability,increased cell apoptosis,increased intracellular ROS content and decreased the mitochondrial membrane potential.Compared with the erastin group,the cell survival rate of erastin+ucOCN group was increased,the level of intracellular ROS was significantly reduced,and the mitochondrial membrane potential was restored.Western blotting results showed that ucOCN could promote the expressions of GPX4 and SLC7A11 proteins in ferroptosis cells.Conclusion ucOCN has an inhibitory effect on erastin⁃induced ferroptosis in HT22 cells.

关 键 词：不全羧化骨钙素 铁死亡 骨源性因子 神经保护 

分 类 号：R967[医药卫生—药理学]