摘要
目的探讨右美托咪定(Dex)对柠檬酸铁铵(FAC)诱导的小鼠海马神经元细胞(HT22)铁超载毒性的保护作用及相关机制。方法选取状态良好的HT22细胞随机分为4组:对照组(Ctrl组)、FAC处理组(FAC组)、Dex处理组(Dex组)、铁死亡抑制剂Fer-1处理组(Fer-1组)。采用FAC诱导细胞制备铁超载模型。随后采用CCK-8法检测HT22细胞的增殖存活率;Western blot检测铁死亡标志性蛋白前列腺素内过氧化物合酶2(PTGS2)、长链酯酰辅酶A合成酶4(ACSL4)、哺乳动物雷帕霉素靶蛋白(mTOR)、转铁蛋白受体1(TFR1)及膜铁输出蛋白(Fpn)的蛋白表达;qPCR检测HT22细胞内PTGS2和ACSL4的基因表达水平;DHE荧光探针检测HT22细胞内的活性氧簇(ROS)水平;MDA检测试剂盒检测HT22细胞内脂质氧化水平;Mito-FerroOrange—亚铁离子探针检测HT22细胞内Fe^(2+)水平;电镜检测细胞内超微结构的改变。结果Dex组和Fer-1组预处理2 h后细胞死亡率明显下降,铁死亡标志物PTGS2和ACSL4蛋白表达和基因表达水平显著降低,细胞超微结构损坏程度明显改善,ROS、脂质氧化以及Fe^(2+)的水平明显低于FAC组(P<0.05)。结论Dex预处理可以减轻FAC诱导的HT22细胞铁超载毒性损伤,与其可能抑制铁死亡有关。
Objective To investigate the protective effect and related mechanisms of dexmedetomidine(Dex)on iron overload toxicity in mouse hippocampal neurons(HT22)induced by ferric ammonium citrate(FAC).Methods Selected HT22 cells in good condition were randomly divided into 4 groups:control group(Ctrl group),FAC treatment group(FAC group),Dex treatment group(Dex group),ferroptosis inhibitor Fer-1 treatment group(Fer-1 group).The iron overload model was established by FAC-induced cells.Subsequently,the proliferation and survival rate of HT22 cells was detected by CCK-8 method;Western blot was used to detect the ferroptosis marker proteins prostaglandin-endoperoxide synthase 2(PTGS2)and acyl-CoA synthetase long-chain family member 4(ACSL4).The protein expressions of mammalian target of rapamycin(mTOR),transferrin receptor 1(TFR1)and ferroportin(Fpn);the gene expression levels of PTGS2 and ACSL4 in HT22 cells were detected by qPCR;Reactive oxygen species(ROS)levels in HT22 cells was detected by DHE fluorescent probe;MDA detection kit was used to detect lipid oxidation levels in HT22 cells;Mito-FerroOrange-ferrous ion probe was used to detect ferrous ion levels in HT22 cells;electron microscopy was used to detect intracellular ultrastructural changes.Results Dex group and Fer-1 group significantly decreased cell death rate after 2 h of pretreatment,the protein and gene expression levels of ferroptosis markers PTGS2 and ACSL4 significantly decreased.The degree of cell ultrastructural damage was significantly improved.The levels of ROS,lipid oxidation and Fe^(2+) were significantly lower than those of the FAC group(P<0.05).Conclusion Dex pretreatment can attenuate FAC-induced iron overload toxicity injury in HT22 cells,which may be related to the inhibition of ferroptosis.
作者
丁慧
王京燕
黄艳
钟薇薇
鲁显福
李元海
Ding Hui;Wang Jingyan;Huang Yan;Zhong Weiwei;Lu Xianfu;Li Yuanhai(Dept of Anesthesiology,Chaohu Hospital Affiliated to Anhui Medical University,Chaohu 238000;Anhui Provincial Laboratory of Inflammatory and Immune Diseases,Anhui Institute of Innovative Drugs,School of Pharmacy,Anhui Medical University,Hefei 230032;Dept of Anesthesiology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)
出处
《安徽医科大学学报》
CAS
北大核心
2022年第10期1633-1639,共7页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:82001185)。
关键词
铁死亡
右美托咪定
铁超载
海马神经元
ferroptosis
dexmedetomidine
iron overload
hippocampal neurons
