常春藤皂苷元通过抑制谷氨酸诱导的铁死亡缓解HT22细胞的炎性反应  

Hederagenin alleviates neuroinflammation response by regulating glutamate-induced ferroptosis in HT22 cells

作　　者：冯雨欣 王贺冉 胡亚卓[2] 孙红美 张小雪 苗秀玲 李姿涵 贾建军[2] Feng Yuxin;Wang Heran;Hu Yazhuo;Sun Hongmei;Zhang Xiaoxue;Miao Xiuling;Li Zihan;Jia Jianjun(Chinese PLA Medical School,Beijing 100853,China)

机构地区：[1]北京解放军医学院,北京100853 [2]解放军总医院第二医学中心老年医学研究所

出　　处：《中华老年心脑血管病杂志》2024年第10期1221-1225,共5页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：国家老年疾病临床研究中心开放课题(NCRCG-PLAGH-2022002);科技部重点研发计划子课题(2018YFC1312301);军队后勤保健课题(21BJZ20);解放军总医院新技术新业务扶持项目(XYZ-202108)。

摘　　要：目的 探讨常春藤皂苷元(hederagenin,HG)对谷氨酸诱导的小鼠海马神经元HT22细胞铁死亡及相应炎性反应的调控作用及其潜在的机制。方法 将HT22细胞分为对照组、谷氨酸组和HG组(n=3),对照组的细胞未接受任何处理,谷氨酸组采用35 mmol/L谷氨酸对HT22细胞进行24 h干预,构建阿尔茨海默病铁死亡的体外模型,HG组使用0.5μmol/L的HG与35 mmol/L谷氨酸共同处理细胞24 h。利用FerroOrange荧光探针测定细胞内Fe^(2+)水平,检测细胞内活性氧水平以及线粒体膜电位的变化,炎性因子肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素(interleukin,IL)-1β、IL-6水平,铁死亡的关键调控蛋白谷胱甘肽过氧化物酶4(glutathione peroxidase4,GPX4)表达。结果 与对照组比较,谷氨酸组细胞内Fe^(2+)、活性氧、TNF-α、IL-1β、IL-6水平明显升高,线粒体膜电位明显降低,差异有统计学意义(P<0.05,P<0.01);与谷氨酸组比较,HG组细胞内Fe^(2+)、活性氧、TNF-α、IL-1β、IL-6水平明显降低,线粒体膜电位明显升高,差异有统计学意义(P<0.05,P<0.01)。与对照组比较,谷氨酸组细胞内GPX4表达明显降低(1.00±0.02 vs 0.46±0.04,P<0.01);与谷氨酸组比较,0.5、1.0μmol/L的HG组细胞内GPX4表达明显升高(0.64±0.03、0.59±0.05 vs 0.46±0.04,P<0.01)。结论 HG通过调节GPX4表达来抑制铁死亡,从而有效减轻炎性反应。Objective To explore the regulatory role of hederagenin(HG) on glutamate(Glu)-induced ferroptosis and corresponding inflammatory responses in mouse hippocampal neuron HT22 cells and investigate its potential mechanisms.Methods HT22 cells were randomly divided into control,Glu and HG groups(n=3).The cells of the control group received no treatment,the cells of the Glu group were treated with 35 mmol/L Glu for 24 h to establish a cellular model of ferroptosis in Alzheimer’s disease,and the cells of the HG group were treated with 0.5 μmol/L HG and 35 mmol/L Glu for 24 h simultaneously.FerroOrange fluorescent probe was used to detect intracellular Fe^(2+).The production of reactive oxygen species(ROS),mitochondrial membrane potential,and levels of inflammatory factors TNF-α,IL-1β and IL-6 in the cells were assessed.Finally,the expression of the key regulator of iron death,glutathione peroxidase 4(GPX4) was measured.Results Compared to the control group,the levels of intracellular Fe^(2+),ROS,TNF-α,IL-1β,and IL-6 were significantly elevated,while the mitochondrial membrane potential was obviously reduced in the Glu group(P<0.05,P<0.01).The HG group had significantly decreased Fe^(2+),ROS,TNF-α,IL-1β,and IL-6 and enhanced mitochondrial membrane potential than the Glu group(P<0.05,P<0.01).The GPX4 expression was significantly lower in the Glu group than the control group(1.00±0.02 vs 0.46±0.04,P<0.01),and was notably higher in the 0.5 and 1.0 μmol/L HG groups when compared to the Glu group(0.64±0.03 and 0.59±0.05 vs 0.46±0.04,P<0.01).Conclusion HG inhibits ferroptosis by regulating GPX4 expression,and thereby effectively alleviates the inflammatory response.

关 键 词：阿尔茨海默病 谷氨酸 炎症 常春藤皂苷元 

分 类 号：R749.16[医药卫生—神经病学与精神病学]