Erythropoietin and Nrf2: key factors in the neuroprotection provided by apo-lactoferrin-猫眼课题宝

DOI：

10.1007/s10534-018-0111-9

发表时间：

2018-05

期刊：

BioMetals

影响因子：

3.5

通讯作者：

中科院分区：

生物学3区

文献类型：

作者：

E. Zakharova;Alex Sokolov;N. Pavlichenko;V. Kostevich;I. Abdurasulova;A. Chechushkov;Irina V. Voynova;A. Elizarova;N. N. Kolmakov-N.;M. G. Bass;Igor Semak;A. Budevich;P. Kozhin;N. Zenkov;V. Klimenko;O. V. Kirik;D. Korzhevskii;E. Menshchikova;V. Vasilyev

乳铁蛋白（LF）具有杀菌、抗贫血、免疫调节、抗肿瘤、抗炎等作用。之前，我们证明了它在体内稳定hif -1- α和hif -2- α的能力，hif -1- α是氧化还原敏感的多目标转录因子。接受重组人LF （rhLF）的动物的各种组织通过表达hif -1- α靶基因而产生反应，从而大量合成了促红细胞生成素（EPO）、铜蓝蛋白等蛋白。发生EPO合成的器官有脑、心、脾、肝、肾和肺。其他研究人员表明，EPO可以作为大鼠严重脑损伤和癫痫持续状态的保护剂。因此，我们尝试将rhLF作为大鼠严重神经系统疾病的保护剂，如鱼藤酮诱导的帕金森病模型和实验性自身免疫性脑脊髓炎作为多发性硬化症模型，并观察到其减轻严重症状的能力。此外，在大脑内侧动脉闭塞1小时后，小鼠腹腔注射rhLF可显著减少缺血第3天脑内坏死面积。在此期间，EPO在小鼠各种组织中合成。已知EPO诱导Nrf2的核易位，Nrf2与hif -1- α一样是一种转录因子。鉴于缺氧条件下两者具有协同保护作用，我们认为LF激活了Keap1/Nrf2信号通路，这是正常细胞和恶性细胞增殖分化的重要环节。J774巨噬细胞在不含铁离子和含铁离子（分别为rmi -1640和DMEM/F12）的情况下培养3天。然后用rhLF或deiprone孵育细胞。共聚焦显微镜显示apo-rhLF（铁-free， rmi -1640）诱导的Nrf2核易位（Keap1/Nrf2信号传导的关键事件）。参比化合物铁螯合剂（deiprone）也有类似的效果。铁结合后（在DMEM/F12中）rhLF不激活Keap1/Nrf2通路。添加到J774中，apo-rhLF增强编码谷胱甘肽s -转移酶P和血红素氧合酶-1的nrf2依赖基因的转录。Western blotting显示，口服载脂蛋白rhlf 6天后，小鼠脑内出现Nrf2，但Fe-rhLF或等量PBS不存在Nrf2。因此，apo-LF，而不是holo-LF，诱导Nrf2从细胞质转位到细胞核，可能是由于其诱导EPO合成的能力。

Among the properties of lactoferrin (LF) are bactericidal, antianemic, immunomodulatory, antitumour, antiphlogistic effects. Previously we demonstrated its capacity to stabilize in vivo HIF-1-alpha and HIF-2-alpha, which are redox-sensitive multiaimed transcription factors. Various tissues of animals receiving recombinant human LF (rhLF) responded by expressing the HIF-1-alpha target genes, hence such proteins as erythropoietin (EPO), ceruloplasmin, etc. were synthesized in noticeable amounts. Among organs in which EPO synthesis occurred were brain, heart, spleen, liver, kidneys and lungs. Other researchers showed that EPO can act as a protectant against severe brain injury and status epilepticus in rats. Therefore, we tried rhLF as a protector against the severe neurologic disorders developed in rats, such as the rotenone-induced model of Parkinson’s disease and experimental autoimmune encephalomyelitis as a model of multiple sclerosis, and observed its capacity to mitigate the grave symptoms. Moreover, an intraperitoneal injection of rhLF into mice 1 h after occlusion of the medial cerebral artery significantly diminished the necrosis area measured on the third day in the ischaemic brain. During this period EPO was synthesized in various murine tissues. It was known that EPO induces nuclear translocation of Nrf2, which, like HIF-1-alpha, is a transcription factor. In view that under conditions of hypoxia both factors demonstrate a synergistic protective effect, we suggested that LF activates the Keap1/Nrf2 signaling pathway, an important link in proliferation and differentiation of normal and malignant cells. J774 macrophages were cultured for 3 days without or in the presence of ferric and ferrous ions (RPMI-1640 and DMEM/F12, respectively). Then cells were incubated with rhLF or Deferiprone. Confocal microscopy revealed nuclear translocation of Nrf2 (the key event in Keap1/Nrf2 signaling) induced by apo-rhLF (iron-free, RPMI-1640). The reference compound Deferiprone (iron chelator) had the similar effect. Upon iron binding (in DMEM/F12) rhLF did not activate the Keap1/Nrf2 pathway. Added to J774, apo-rhLF enhanced transcription of Nrf2-dependent genes coding for glutathione S-transferase P and heme oxygenase-1. Western blotting revealed presence of Nrf2 in mice brain after 6 days of oral administration of apo-rhLF, but not Fe-rhLF or equivalent amount of PBS. Hence, apo-LF, but not holo-LF, induces the translocation of Nrf2 from cytoplasm to the nucleus, probably due to its capacity to induce EPO synthesis.

Erythropoietin and Nrf2: key factors in the neuroprotection provided by apo-lactoferrin

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