摘要
目的探讨环指蛋白13(ring finger protein 13,RNF13)在脑缺血再灌注损伤(cerebral ischemia reperfusion injury,CIRI)中对神经元铁死亡的调控作用及分子机制。方法以小鼠大脑中动脉栓塞术和原代神经元氧糖剥夺再复氧处理建立疾病模型,使用CRISPR/Cas9基因敲除技术、免疫组化染色、免疫荧光染色、Westen blot和脂质过氧化检测等方法评估RNF13在CIRI中对铁死亡的调控作用及分子机制。两样本均数比较采用t检验,多样本均数比较采用单因素方差分析。结果小鼠和原代神经元造模后,RNF13蛋白水平均高于对照组(均P<0.05)。敲除RNF13通过下调铁蛋白重链1(ferritinheavychain1,Fth1)和铁蛋白轻链(ferritin light chain,Ftl)(均P<0.05)促进小鼠脑组织中的游离亚铁离子含量和脂质过氧化物累积(均P<0.01),促进铁死亡,最终导致神经功能缺损(P<0.01)。结论RNF13减轻CIRI后的神经元铁死亡,该作用依赖于Fthl。
Objective To investigate the function of Ring finger protein 13(RNF13)in cerebral ischemia reperfusion injury(CIRI),and its mechanism.Methods The mouse middle cerebral artery embolization and primary neuron oxygen-glucose deprivation and reoxygenation were used as disease models.The CRISPR/Cas9 gene knockout technique,immunohistochemical staining,immunofluorescence staining,Western blot and lipid peroxidation detection were used to evaluate the regulatory effect and molecular mechanism of RNF13 on ferroptosis in CIRI.Student's t test was used for the comparison of two samples,and oneway analysis of variance was used for the comparison of multiple samples.Results The expression levels of RNF13 protein in mice and primary neurons were upregulated during CIRI.After knockout of RNF13,ferritin heavy chain 1(Fth1)and ferritin light chain(Ftl)were downregulated,and the content of free ferrous ions and the accumulation of lipid peroxides in mice brain tissues were promoted,leading to ferroptosis aggravation and neurological impairments.Overexpression of RNF13 protected against ferroptosis by reducing the production of free ferrous and lipid peroxides in neurons.Conclusion RNF13 alleviates ferroptosis in neurons after CIRI,and the effect is induced by Fthl.
作者
李爽
王强平
周文科
郑保平
刘强
李震东
张方成
王海均
Li Shuang;Wang Qiangping;Zhou Wenke(Department of Neurosurgery,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022,China;Department of Neurosurgery,theFirst Af filiated Hospital of Xinziang Medical University,Xinziang 453100China)
出处
《华中科技大学学报(医学版)》
北大核心
2025年第4期459-464,共6页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
国家自然科学基金资助项目(No.81201026)
湖北省自然科学基金资助项目(No.2021CFB549)。
关键词
脑缺血再灌注损伤
铁死亡
环指蛋白13
神经元
铁蛋白
cerebral ischemia-reperfusion injury
ferroptosis
ring finger protein 13
neuron
ferritin
作者简介
李爽,男,1997年生,硕士研究生,E-mail:lishuang2312@126.com;通信作者:王海均,E-mail:whjl1260@163.com。
