摘要
目的研究血管性痴呆(VD)中铁死亡的作用与机制。方法选取36只体质量200~220 g的健康雄性SD大鼠,采用随机数字表法将大鼠分为3组:(1)假手术组:仅分离但不结扎血管;(2)VD组:采用改良双侧颈总动脉永久结扎法制作VD模型大鼠;(3)Ferrostatin-1(Fer-1)组:制备VD模型大鼠后,予以特异性铁死亡抑制剂Fer-1腹腔内注射。VD模型造模成功1个月后,通过Morris水迷宫试验评价大鼠的认知功能,采用尼氏染色观察大鼠海马病理改变,使用Perls’铁染色检测海马铁沉积情况,电镜下观察大鼠海马CA1区神经元超微结构,应用RT-PCR检测大鼠海马前列腺素内过氧化物合酶2(PTGS2)mRNA的表达水平,使用ELISA法检测海马丙二醛(MDA)和谷胱甘肽(GSH)的表达水平,运用Western blot检测海马4-羟基壬烯醛(4-HNE)和谷胱甘肽过氧化物酶4(GPX4)的表达水平。结果与假手术组相比,VD模型大鼠逃逸潜伏期延长(P<0.05),跨越平台次数及靶象限停留时间均减少(P<0.05),大鼠海马CA1区神经元数量减少(P<0.05),神经元线粒体皱缩,铁沉积增多(P<0.05);海马PTGS2 mRNA表达增多(P<0.05),GSH及GPX4表达下调(P<0.05),MDA及4-HNE表达增多(P<0.05)。与VD模型大鼠相比,腹腔注射Fer-1后大鼠认知功能显著改善,海马CA1区神经元数量增多(P<0.05),铁沉积减少(P<0.05);海马PTGS2 mRNA表达增加(P<0.05),GPX4和GSH表达上调(P<0.05),4-HNE和MDA表达下调(P<0.05)。结论铁死亡参与VD的发生发展,Fer-1通过减少铁沉积、增强抗氧化能力及减少脂质过氧化物堆积等抑制海马神经元铁死亡,改善VD模型大鼠的认知功能。
Objective To investigate the potential function and mechanism of ferroptosis in the pathogenesis of VD.MethodsThirty-six healthy male SD rats weighing 200-220 g were selected and randomly divided into three groups using random number table method:(1)Sham operation group,the carotid arteries were separated but not ligated.(2)VD group:the VD rat model was established by modified ligation of bilateral common carotid arteries(2-VO).(3)Ferrostatin-1(Fer-1)group:after VD rat model was established,specific inhibitor of ferroptosis Fer-1 was injected intraperitoneally. One month after 2-VO operation,the Morris water maze test was used to measure rats.intelligence. The pathological changes in the hippocampus were examined via Nissl staining. Perls.staining was used to detect iron accumulation. The ultrastructure of neurons in the hippocampal CA1 area was examined by transmission electron microscopy.The expression level of prostaglandin-endoperoxide synthase(PTGS2)mRNA was detected by RT-PCR. The levels of malondialdehyde(MDA)and glutathione(GSH)were measured by ELISA. Protein expression levels of 4-hydroxynonenal(4-HNE)and glutathione peroxidase 4(GPX4)were determined by Western blot.Results was prolonged(P<0.05),and times of crossing platform and time spent in the target quadrant decreased(P<0.05). In the hippocampal CA1 region of VD rats,decreased number of neurons(P<0.05),shrinkage of neuronal mitochondria,and increased deposition of iron(P<0.05)were observed. Expression of PTGS2 m RNA increased(P<0.05). Expression levels of GSH and GPX4 decreased(P<0.05),while expression levels of MDA and4-HNE increased(P<0.05)in VD rat models. Compared with VD rat models,the cognitive function of rats improved significantly after intraperitoneal injection of Fer-1. The number of neurons in hippocampal CA1 area increased(P<0.05). Administration with Fer-1 upregulated expression levels of GPX4 and GSH,as well as reduced expression levels of 4-HNE and MDA,iron accumulation,and PTGS2 mRNA in the hippocampus(P<0.05).ConclusionFerroptosis
作者
贺莎莎
刘进友
李书剑
田发发
方佳
HE Shasha;LIU Jinyou;LI Shujian;TIAN Fafa;FANG Jia(The Second Xiangya Hospital,Central South University,Changsha 410011,China;Henan Provincial People's Hospital,Zhengzhou 450003,China;Xiangya Hospital,Central South University,Changsha 410008,China)
出处
《中国实用神经疾病杂志》
2021年第15期1289-1298,共10页
Chinese Journal of Practical Nervous Diseases
基金
国家自然科学基金(编号:81702582,81902550)
湖南省自然科学基金(编号:S2017JJQNJJ2112)。
