丁基苯酞通过抑制细胞焦亡和铁死亡减轻脂多糖诱导的小鼠认知功能障碍  

Dl-3-n-butylphthalide alleviates lipopolysaccharide-induced cognitive dysfunction in mice by inhibiting pyroptosis and ferroptosis

作　　者：霍康[1] 马凯歌 段鹏[1] 王建懿 卫萌[1] 张萌[1] 许静[1] HUO Kang;MA Kaige;DUAN Peng(The First Affiliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)

机构地区：[1]西安交通大学第一附属医院,陕西西安710061 [2]西安交通大学基础医学院神经生物学研究所

出　　处：《精神医学杂志》2022年第5期449-457,共9页Journal of Psychiatry

基　　金：陕西省自然科学基础研究计划项目(编号:2023-JC-YB-736,2016JM8019);西安市科技计划项目(编号:2016048SF/YX04(2));西安交通大学第一附属医院基金自由探索项目(编号:2019ZYTS-10)。

摘　　要：目的探讨丁基苯酞(NBP)对脂多糖(LPS)诱导的小鼠认知功能障碍(CD)的治疗作用及机制。方法将C57BL/6J小鼠按照干预措施的不同随机分为5组(n=12):对照组、LPS组、LPS+NBP组、LPS+NBP+TMAO(细胞焦亡诱导剂)组和LPS+NBP+GPX4-IN-3(铁死亡诱导剂)组。各组小鼠分别使用相应药物干预7 d。通过Morris水迷宫实验评价小鼠的认知功能。使用商用试剂盒检测小鼠血清中白细胞介素(IL)-1β、IL-6和肿瘤坏死因子-α(TNF-α)水平及海马组织中丙二醛(MDA)、超氧化物歧化酶(SOD)和活性氧(ROS)的水平,并对小鼠海马组织进行了尼氏染色和Iba-1免疫荧光染色。使用Iron Assay试剂盒测定海马组织Fe^(2+)含量。通过Western blot检测海马组织中核苷酸结合寡聚化结构域样受体3(NALP3)、caspase-1、谷胱甘肽过氧化物酶4(GPX4)和前列腺素内过氧化物合酶2(PTGS2)的蛋白表达。结果与LPS组相比,LPS+NBP组小鼠的逃避潜伏期减少,穿越平台次数增加(P<0.05)。与LPS组相比,LPS+NBP组小鼠的血清IL-1β、IL-6和TNF-α水平降低(P<0.05),海马组织中的Iba-1相对荧光强度降低(P<0.05)。与LPS组相比,LPS+NBP组小鼠海马组织中MDA和ROS水平降低,SOD升高(P<0.05)。与LPS组相比,LPS+NBP组小鼠海马组织尼氏光密度升高(P<0.05)。与LPS组相比,LPS+NBP组小鼠海马组织NALP3和caspase-1的蛋白相对表达量降低(P<0.05)。与LPS组相比,LPS+NBP组小鼠海马组织Fe^(2+)含量和PTGS2的蛋白相对表达量降低,GPX4的蛋白相对表达量升高(P<0.05)。TMAO和GPX4-IN-3的使用逆转了NBP对上述指标的影响(P<0.05)。结论NBP可减轻外周炎症诱导的小鼠认知功能障碍,NBP至少部分通过抑制细胞焦亡和铁死亡来减轻小鼠认知功能障碍。Objective To explore the therapeutic effect and mechanism of dl-3-n-butylphthalide(NBP)on lipopolysaccharide(LPS)-induced cognitive dysfunction(CD)in mice.Methods C57BL/6J mice were randomly divided into 5 groups(n=12)according to the different intervention measures:control group,LPS group,LPS+NBP group,LPS+NBP+TMAO(pyroptosis inducer)group and LPS+NBP+GPX4-IN-3(ferroptosis inducer)group.Mice in LPS group were intraperitoneally injected with 0.1 ml 0.8 mg/kg LPS,1 h followed by intraperitoneal injection of 0.1 ml 10%dimethyl sulfoxide+90%corn oil mixture.Mice in LPS+NBP group were intraperitoneally injected with 0.1 ml 0.8 mg/kg LPS,1 h followed by intraperitoneal injection of 0.1 ml 20 mg/kg NBP.Mice in LPS+NBP+TMAO group were intraperitoneally injected with 0.1 ml 0.8 mg/kg LPS,1 h followed by intraperitoneal injection of 0.1 ml 20 mg/kg NBP.On this basis,the TMAO was dissolved in water at a concentration of 1.5%for mice to drink.Mice in LPS+NBP+GPX4-IN-3 group were intraperitoneally injected with 0.1 ml 0.8 mg/kg LPS,1 h followed by intraperitoneal injection of 0.1 ml 20 mg/kg NBP and 0.1 ml 15 mg/kg GPX4-IN-3(GPX4-IN-3 dissolved in 10%dimethyl sulfoxide+90%corn oil).Mice in control group were intraperitoneally injected with 0.1 ml saline,1 h followed by intraperitoneal injection of 0.1 ml 10%dimethyl sulfoxide+90%corn oil mixture.Mice in each group were treated with corresponding drugs for 7 days.The cognitive function of mice was evaluated by Morris water maze test.The levels of interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)in serum and levels of malondialdehyde(MDA),superoxide dismutase(SOD)and reactive oxygen species(ROS)in hippocampus of mice were detected using commercial kits,and Nissl staining and Iba-1 immunofluorescence staining were performed on the hippocampus of mice.Fe^(2+)content in hippocampus was determined through Iron Assay kit.The protein expressions of nucleotide-binding oligomerization domain-like receptor 3(NALP3),caspase-1 and glutathione peroxidase 4(GPX4)and prostagland

关 键 词：丁基苯酞 认知功能障碍 外周炎症 细胞焦亡 铁死亡 

分 类 号：R742[医药卫生—神经病学与精神病学]