背景:小胶质细胞在脑出血后神经损伤中起关键作用。铁代谢,ICH后失调,参与小胶质细胞功能障碍。先前的研究表明,与铁代谢相关的脂质运载蛋白2(LCN2)参与ICH后小胶质细胞功能的调节。在这项研究中,我们研究了LCN2在ICH后小胶质细胞功能中的作用.
方法:BV2(小胶质细胞)细胞系,用LCN2转染过表达/干扰,在体外接受C57BL/6小鼠的血液输注。对于LCN2功能的体内研究,在小鼠中进行LCN2敲除。Leproxstatin-1和RSL3用于操纵铁凋亡并研究LCN2对ICH后小胶质细胞的影响。BV2(小胶质细胞)细胞系,用铁蛋白轻链(FTL)转染过表达/干扰,与原代培养的神经元共培养,研究LCN2的作用机制。行为测试在ICH前和ICH后第3、7和28天进行,收集大脑和培养细胞的蛋白质,组织学,和形态学研究。
结果:小胶质细胞中LCN2的表达上调,星形胶质细胞,和神经元,并在ICH后发挥危险作用。在小胶质细胞中,LCN2促进铁凋亡,这促进了ICH后的神经损伤。LCN2介导的FTL缺陷被证明是小胶质细胞铁凋亡诱导的神经损伤的原因。
结论:我们的研究表明,在ICH后,LCN2增强的小胶质细胞铁性凋亡通过诱导FTL缺乏发挥有害作用。目前的研究揭示了参与ICH病理生理进程的新分子机制。
BACKGROUND: Microglia play pivotal roles in post-intracerebral hemorrhage (ICH) neural injury. Iron metabolism, which is dysregulated after ICH, participates in microglial dysfunction. Previous studies have shown that iron metabolism-related lipocalin-2 (LCN2) is involved in regulating microglial function following ICH. In this study, we investigated the role of LCN2 in microglial function following ICH.
METHODS: The BV2 (microglia) cell line, transfected with LCN2 for overexpression/interference, received a blood infusion from C57BL/6 mice in vitro. For the in vivo study of LCN2 function, an LCN2 knockout was conducted in mice. Liproxstatin-1 and RSL3 were used to manipulate ferroptosis and to study the effects of LCN2 on microglia after ICH. A BV2 (microglia) cell line, transfected with ferritin light chain (FTL) for overexpression/interference, was co-cultured with primary cultured neurons for a study on the mechanism of LCN2. Behavioral tests were conducted pre-ICH and on days 3, 7, and 28 post-ICH, and the brains and cultured cells were collected for protein, histological, and morphological studies.
RESULTS: Brain LCN2 expression was upregulated in microglia, astrocytes, and neurons and played hazardous roles after ICH. In microglia, LCN2 promoted ferroptosis, which facilitated neural injury after ICH. LCN2-mediated FTL deficiency was shown to be responsible for microglial ferroptosis-induced neural injury.
CONCLUSIONS: Our study suggests that LCN2-enhanced microglial ferroptosis plays a detrimental role by inducing FTL deficiency after ICH. The current study reveals a novel molecular mechanism involved in the pathophysiological progression of ICH.