艾司氯胺酮减轻HT22细胞炎症损伤及铁死亡相关机制  

Esketamine reduces inflammatory damage in HT22 cells and ferroptosis mechanisms

作　　者：王京燕 丁慧 钟薇薇[2] 鲁显福[2] 黄艳[3] 李元海[1,2] WANG Jing-yan;DING Hui;ZHONG Wei-wei;LU Xian-fu;HUANG Yan;LI Yuan-hai(Dept of Anesthesiology,Chaohu Hospital Affiliated to Anhui Medical University,Chaohu,Anhui 238000,China;Dept of Anesthesiology,the First Affiliated Hospital of Anhui Medical University,Anhui Medical University,Hefei 230032,China;School of Pharmacy,Anhui Medical University,Hefei 230032,China)

机构地区：[1]安徽医科大学附属巢湖医院麻醉科,安徽巢湖238000 [2]安徽医科大学第一附属医院麻醉科,安徽合肥230032 [3]安徽医科大学药学院,安徽合肥230032

出　　处：《中国药理学通报》2022年第11期1647-1654,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No.82001185)。

摘　　要：目的探讨艾司氯胺酮(esketamine,ESK)对脂多糖(lipopolysaccharide,LPS)诱导的HT22细胞(小鼠海马神经元细胞)损伤以及相关机制。方法1μmol·L^(-1) ESK预处理2 h,再经过10 mg·L^(-1) LPS处理24 h后,Western blot检测高迁移率族蛋白B1(high mobility group box-1,HMGB1)、长链酰基辅酶A合成酶4(long chain acyl CoA synthetase 4,ACSL4)、前列素内环氧化物合成酶2(prostaglandin-endoperoxide synthase 2,PTGS2)、转铁蛋白受体1(transferrin receptor 1,TFR1)、铁转运蛋白(ferroportin,FPN)和铁蛋白(ferritin)表达;ELISA检测白介素-1β(interleukin-1beta,IL-1β)和肿瘤坏死因子-α(necrosis factor alpha,TNF-α)表达;DHE荧光探针检测细胞内活性氧(reactive oxygen species,ROS)改变;MDA试剂盒检测细胞内脂质过氧化水平;FerroOrange荧光探针检测细胞内二价铁离子水平;电镜观察线粒体超微结构改变。结果与正常组相比,LPS组细胞ROS、脂质过氧化及Fe^(2+)水平明显升高;ACSL4、PTGS2、HMGB1、TFR1、FPN蛋白水平表达明显上升,铁蛋白减少;线粒体嵴肿胀,出现外膜破裂。与LPS组相比,ESK预处理组ROS、脂质过氧化及Fe^(2+)水平明显降低;ACSL4、PTGS2、HMGB1、TFR1和FPN蛋白水平明显下降,铁蛋白降解减少;线粒体嵴肿胀减轻。结论ESK可能通过调节HMGB1表达来抑制铁死亡,进而减轻LPS诱导的神经细胞损伤。Aim To investigate the effect of esketamine(ESK)on lipopolysaccharide(LPS)-induced damage to HT22 cells(mouse hippocampal neuron cells)and the underlying mechanism.Methods After ESK pretreatment for 2 h and then LPS treatment for 24 h,Western blot was used to detect high mobility group box-1(HMGB1),long chain acyl CoA synthetase 4(ACSL4),prostaglandin-endoperoxide synthase 2(PTGS2),transferrin receptor 1(TFR1),ferroportin(FPN)and ferritin expression.ELISA was used to detect the expression of inflammatory protein interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α).DHE fluorescent was probed to detect the changes in intracellular reactive oxygen species(ROS).MDA kit was used to detect intracellular lipid oxidation levels.FerroOrange fluorescent was probed to detect intracellular divalent iron ion levels.Electron microscopy was used to observe the changes in mitochondrial ultrastructure.Results Compared with the normal group,the levels of ROS,lipid peroxidation and Fe^(2+)in cells of LPS group significantly increased,the expression of ACSL4,PTGS2,HMGB1,TFR1 and FPN proteins increased significantly,ferritin decreased as well,and the mitochondrial crest was swollen and the outer membrane rupture occurred.Compared with LPS group,ROS,lipid peroxidation and Fe^(2+)levels of ESK pretreatment group decreased significantly;The protein levels of ACSL4,PTGS2,HMGB1,TFR1 and FPN decreased significantly,and the degradation of ferritin decreased;and the swelling of mitochondrial crest decreased.Conclusion ESK may inhibit ferroptosis by regulating HMGB1 expression,thereby alleviating LPS-induced nerve cell damage.

关 键 词：艾司氯胺酮 铁死亡 HMGB1 神经炎症 ROS 脂质过氧化 

分 类 号：R-332[医药卫生] R322.81R364.5R59.1R971.2