Nrdp1 在慢性间歇性低氧诱导的大鼠脑损伤中的作用通过调节 ErbB3 蛋白水平实现。

Role of the Nrdp1 in Brain Injury Induced by Chronic Intermittent Hypoxia in Rats via Regulating the Protein Levels of ErbB3.

机构信息

Department of Emergency, Guizhou Provincial People's Hospital, Guiyang, China.

Department of Emergency, Affiliated Hospital of Guizhou Medical University, Guiyang, China.

出版信息

Neurotox Res. 2020 Jun;38(1):124-132. doi: 10.1007/s12640-020-00195-z. Epub 2020 Mar 21.

DOI:10.1007/s12640-020-00195-z

PMID:32200526

Abstract

Obstructive sleep apnea syndrome (OSAS) is known as a repeated obstruction of the upper airway during sleep, leading to generalized hypoxia episodes and associated with cardiovascular and cerebrovascular diseases. We mainly explored the role of neuregulin receptor degradation protein-1 (Nrdp1, also known as FLRF) in brain injury induced by chronic intermittent hypoxia (CIH) in rats. Wistar rats were randomly divided into 4 groups (n = 12 per group), including the sham + adeno-associated virus-NC (AAV-NC) group, the sham + AAV-siNrdp1 group, the IH-4w (intermittent hypoxia for 4 weeks) + AAV-NC group, and the IH-4w + AAV-siNrdp1 group. Morphologic changes in brain tissue were observed by hematoxylin and eosin (HE) staining. Apoptosis in the hippocampus was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining. Spatial learning and memory were assessed by the Morris water maze test. The expression of Nrdp1 mRNA and protein in the hippocampus was detected by qualitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. The concentration of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in serum was detected via enzyme-linked immunosorbent assay (ELISA) kits. Nrdp1 expression was increased after intermittent hypoxia exposure over time. Western blotting and H&E results showed that pathological changes of hippocampus neurons in chronic intermittent hypoxia rat were diminished by shNrdp1. Western blotting and TUNEL staining showed that apoptotic cells in the hippocampus of CIH rats were decreased by shNrdp1. The Morris water maze results proved that shNrdp1 improved spatial learning performance of chronic intermittent hypoxia rats. ELISA kits results showed that CIH-induced inflammatory response was decreased by shNrdp1. Western blotting and qRT-PCR results showed protein expression of ErbB3 in the hippocampus of CIH rats. Nrdp1 could regulate ErbB3 protein levels in brain-injured rats with CIH, which demonstrates that Nrdp1 is a potential therapeutic target in the cognition deficits associated with OSAS.

摘要

阻塞性睡眠呼吸暂停综合征（OSAS）是一种在睡眠期间上气道反复阻塞的疾病，导致全身缺氧发作，并与心血管和脑血管疾病相关。我们主要探讨了神经调节蛋白受体降解蛋白 1（Nrdp1，也称为 FLRF）在慢性间歇性低氧（CIH）诱导的大鼠脑损伤中的作用。Wistar 大鼠随机分为 4 组（每组 12 只），包括假手术+腺相关病毒-NC（AAV-NC）组、假手术+AAV-siNrdp1 组、4 周间歇性低氧（IH-4w）+AAV-NC 组和 4 周间歇性低氧（IH-4w）+AAV-siNrdp1 组。通过苏木精和伊红（HE）染色观察脑组织形态变化。通过末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记（TUNEL）染色检测海马区细胞凋亡。通过 Morris 水迷宫试验评估空间学习和记忆能力。通过定性实时聚合酶链反应（qRT-PCR）和 Western blot 检测海马区 Nrdp1 mRNA 和蛋白的表达。通过酶联免疫吸附试验（ELISA）试剂盒检测血清中白细胞介素 6（IL-6）和肿瘤坏死因子-α（TNF-α）的浓度。随着间歇性低氧暴露时间的延长，Nrdp1 的表达增加。Western blot 和 HE 结果表明，shNrdp1 减轻了慢性间歇性低氧大鼠海马神经元的病理变化。Western blot 和 TUNEL 染色显示，shNrdp1 减少了 CIH 大鼠海马区的凋亡细胞。Morris 水迷宫结果表明，shNrdp1 改善了慢性间歇性低氧大鼠的空间学习能力。ELISA 试剂盒结果表明，shNrdp1 降低了 CIH 诱导的炎症反应。Western blot 和 qRT-PCR 结果显示，CIH 大鼠海马区 ErbB3 蛋白表达增加。Nrdp1 可调节 CIH 脑损伤大鼠的 ErbB3 蛋白水平，表明 Nrdp1 是 OSAS 相关认知障碍的潜在治疗靶点。