黄海锋,高玉元,段青蕊,王丽娟

• 1:南方医科大学附属广东省人民医院(广东省医学科学院)神经科

摘要(Abstract)：

目的 探究N-myc下游调控基因1(N-myc downstream regulated gene 1,NDRG1)对氧糖剥夺(oxygen-glucose deprivation, OGD)诱导的小鼠脑微血管内皮细胞(brain microvascular endothelial cells, BMVEC)铁死亡的影响。方法 提取并培养小鼠原代BMVEC,电镜观察氧糖剥夺后原代BMVEC线粒体形态。体外培养bEnd.3细胞，分为对照组、模型组(构建OGD模型)、小干扰RNA阴性对照(si-NC)组、NDRG1干扰RNA(si-NDRG1)组、阴性对照造模(OGD+si-NC)组、NDRG1干扰造模(OGD+si-NDRG1)组(n=3)。模型组、OGD+si-NC组和OGD+si-NDRG1组构建OGD模型，si-NC组和OGD+si-NC组进行si-NC转染，si-NDRG1组和OGD+si-NDRG1组进行si-NDRG1转染，检测各组细胞活力、丙二醛、谷胱甘肽、Fe~(2+)水平、脂质过氧化水平、NDRG1、谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4)及酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long-chain family member 4,ACSL4)蛋白水平。结果 与对照组比较，模型组OGD处理后的细胞贴壁数量显著减少，胞质出现肿胀现象，细胞活力显著降低[(37.68±2.43)%vs(96.34±12.08)%,P<0.05],GPX4明显下调(0.78±0.02 vs 1.15±0.01,P<0.01),ACSL4(1.45±0.04 vs 0.78±0.12,P<0.01)、NDRG1蛋白表达明显上调(1.22±0.01 vs 0.13±0.01,P<0.01)。si-NDRG1组GPX4、NDRG1蛋白表达明显低于si-NC组，ACSL4、谷胱甘肽蛋白表达明显高于si-NC组，差异有统计学意义(P<0.05)。OGD+si-NC组GPX4蛋白表达及谷胱甘肽水平明显低于si-NC组和si-NDRG1组，NDRG1、ACSL4蛋白表达、丙二醛、Fe~(2+)相对荧光强度、氧化型脂质相对荧光强度明显高于si-NC组和si-NDRG1组，差异有统计学意义(P<0.05)。OGD+si-NDRG1组GPX4蛋白表达及谷胱甘肽水平明显低于si-NC组和si-NDRG1组，NDRG1明显低于si-NC组，明显高于si-NDRG1组，ACSL4、Fe~(2+)相对荧光强度明显高于si-NC组和si-NDRG1组(P<0.05)。OGD+si-NDRG1组NDRG1、ACSL4、丙二醛、Fe~(2+)相对荧光强度、氧化型脂质相对荧光强度明显低于OGD+si-NC组，差异有统计学意义(P<0.05)。结论 降低NDRG1可以通过改善铁代谢和脂质过氧化，减轻OGD诱导的微血管内皮细胞铁死亡。

关键词(KeyWords)： 卒中;铁死亡;内皮细胞;缺氧缺血,脑;基因,myc

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81974195,82101377,82371249,82471257);; 广东省科技计划项目(2020A0505140006);; 广州市科技计划项目(202201000005);; 广东医学科学技术研究基金项目(A2021308,A2022092)

作者(Author): 黄海锋,高玉元,段青蕊,王丽娟

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