PKR inhibitor protects spinal cord injury through mitigating endoplasmic reticulum stress and pyroptosis

IF 4.4 3区医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Neurochemistry international Pub Date : 2023-10-20 DOI:10.1016/j.neuint.2023.105632

Ze Yang , Ming Sheng , Meng Wang , Long Cheng , Xin Sun

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Abstract

Objectives

The goal of the study was to reveal the regulatory role of protein kinase R (PKR) in spinal cord injury (SCI), a devasting disorder of the neurological system, and to elucidate its potential mechanism.

Methods

The established animal and cellular models of SCI were treated by the PKR inhibitor C12. Histological injury and tissue apoptosis were assessed via H&E staining and TUNEL assays, respectively. Basso-Beattie-Bresnahan (BBB) scoring as well as forelimb grip strength tests were employed to evaluate functional recovery. The production of ROS and cytokines were appraised via their related commercial kits. Western blot and immunofluorescence assay were used to examine protein expression. CCK-8 method was used to assay cell activity. Co-immunoprecipitation assay was conducted to measure the affinity of PKR with STAT1.

Results

PKR expression was enhanced following SCI, and the PKR inhibitor C16 mitigated histological injury, cell apoptosis and water content in spinal cord, and improved function recovery following SCI. Meanwhile, C16 attenuated ER stress, pyroptosis, NLRP3 inflammasome and inflammation in mice with SCI and in BV-2 cells challenged with LPS. Additionally, PKR interacted with STAT1 in BV-2 cells, and STAT1 knockdown inhibited ER stress, pyroptosis and inflammation in BV-2 cells challenged with LPS. The protective role of C16 in BV-2 cells exposed to LPS were partly abolished by STAT1 overexpression.

Conclusion

PKR inhibition might be a prospective effective approach to attenuating SCI and accelerating function recovery through modulating microglial pyroptosis and ER stress.

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PKR抑制剂通过减轻内质网应激和焦下垂来保护脊髓损伤。

目的：本研究旨在揭示蛋白激酶R（PKR）在脊髓损伤（SCI）中的调节作用，并阐明其潜在机制。方法：采用PKR抑制剂C12对建立的SCI动物和细胞模型进行治疗。组织学损伤和组织凋亡分别通过H&E染色和TUNEL测定进行评估。Basso Beattie Bresnahan（BBB）评分以及前肢握力测试用于评估功能恢复。ROS和细胞因子的产生通过其相关的商业试剂盒进行评估。采用蛋白质印迹和免疫荧光法检测蛋白质表达。采用CCK-8法测定细胞活性。采用免疫共沉淀法测定PKR与STAT1的亲和力。结果：SCI后PKR表达增强，PKR抑制剂C16减轻了组织学损伤、细胞凋亡和脊髓含水量，改善了SCI后的功能恢复。同时，C16减轻了SCI小鼠和BV-2小鼠的ER应激、pyroptosis、NLRP3炎症小体和炎症 用LPS攻击的细胞。此外，PKR与BV-2中的STAT1相互作用 细胞和STAT1敲除抑制BV-2的ER应激、pyroptosis和炎症 用LPS攻击的细胞。C16在BV-2中的保护作用 暴露于LPS的细胞被STAT1过表达部分消除。结论：抑制PKR可能是一种通过调节小胶质细胞焦下垂和ER应激来减轻SCI和加速功能恢复的前瞻性有效方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Neurochemistry international 医学-神经科学

CiteScore

8.40

自引率

2.40%

发文量

128

审稿时长

37 days

期刊介绍： Neurochemistry International is devoted to the rapid publication of outstanding original articles and timely reviews in neurochemistry. Manuscripts on a broad range of topics will be considered, including molecular and cellular neurochemistry, neuropharmacology and genetic aspects of CNS function, neuroimmunology, metabolism as well as the neurochemistry of neurological and psychiatric disorders of the CNS.