蛋白激酶C相互作用蛋白1基因敲除诱导神经元氧化应激  

Knockout of protein kinase C interacting protein 1 gene induces neuronal oxidative stress

作　　者：马乾凤[1] 侯晓霖[1] 陈秋元 李艳宁 王俊燕 潘晓玥 宿瑞琼 雷哲 王银[2] 李云鸿[2] MA Qianfeng;HOU Xiaolin;CHEN Qiuyuan;LI Yanning;WANG Junyan;PAN Xiaoyue;SU Ruiqiong;LEI Zhe;WANG Yin;LI Yunhong(General Hospital of Ningxia Medical University,Yinchuan 750004,China;Department of Physiology and Neurobiology,School of Basic Medical Sciences,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院超声科,宁夏银川750004 [2]宁夏医科大学基础医学院生理学与神经生物学系,宁夏银川750004

出　　处：《宁夏医学杂志》2024年第11期925-928,F0002,共5页Ningxia Medical Journal

基　　金：宁夏自然科学基金项目(2023AAC03547,2023AAC03162,2023AAC02037);宁夏重点研发计划项目(2021BEG03097);宁夏回族自治区卫生健康委员会适宜技术推广项目(2022-NWSY-015)。

摘　　要：目的 研究蛋白激酶C相互作用蛋白1(PICK1)敲除对神经元氧化应激反应的影响,从而明确PICK1在神经元氧化应激反应中的作用。方法 剪取小鼠尾巴1 cm,匀浆后提取DNA和蛋白质,利用PCR和Western blot鉴定野生型小鼠和PICK1敲除鼠的基因型。利用ELISA检测试剂盒,检测野生型和PICK1敲除小鼠脑组织中活性氧簇(ROS)和谷胱甘肽(GSH)的水平。硝基酪氨酸(Nitrotyrosine)是氧化损伤标志因子,利用Western blot及免疫荧光染色方法,检测小鼠脑内Nitrotyrosine与神经元的标记物MAP2。结果 成功鉴定出野生型鼠、PICK1基因杂合子小鼠和纯合子小鼠。PICK1敲除小鼠脑内的氧化应激因子ROS (P<0.05)和Nitrotyrosine水平显著升高(P<0.05),而抗氧化应激因子GSH明显下降(P<0.05)。Nitrotyrosine与MAP2免疫荧光共染结果显示,PICK1敲除的小鼠中神经元细胞存在氧化损伤。结论 PICK1基因敲除导致了小鼠神经元的氧化应激反应,提示PICK1可能是抑制神经元氧化应激的一个靶点。Objective To investigate the effect of protein kinase C interacting protein 1(PICK1)knockout on neuronal oxidative stress response,and to clarify the role of PICK1 in neuronal oxidative stress response.Methods The mouse tail of 1cm was trimmed and homogenized,then DNA and proteins were extracted,and the genotypes of wild-type mice and PICK1 knockout mice were identified using PCR and western blot.The level of reactive oxygen species(ROS)was detected by ELISA and glutathione(GSH)assay kit was used to exam GSH level.Western blot was operated to check the expression level of oxidative damage indicator Nitrotyrosine,and immunofluorescence staining was done to detect the co-location of Nitrotyrosine and MAP2,the marker of neurons in the mouse brain.Results The wild-type mice,PICK1 gene heterozygous mice and homozygous mice were successfully identified.The levels of oxidative stress factors ROS(P<0.05)and Nitrotyrosine in the brain of PICK1 knockout mice significantly increased(P<0.05),while the levels of antioxidant stress factor GSH significantly decreased(P<0.05).The immunofluorescence co-staining results of Nitrotyrosine and MAP2 showed oxidative damage to neuronal cells in PICK1 knockout mice.Conclusion Knockout of PICK1 gene could lead to oxidative stress response in mouse neurons and suggested that PICK1 may be a target for inhibiting neuronal oxidative stress.

关 键 词：蛋白激酶C相互作用蛋白1 神经元 氧化应激 基因敲除 

分 类 号：R739[医药卫生—肿瘤]