抑制内质网应激通过减少 NLRP3 炎性小体激活介导的神经炎症来减轻围手术期神经认知障碍。

Inhibiting endoplasmic reticulum stress alleviates perioperative neurocognitive disorders by reducing neuroinflammation mediated by NLRP3 inflammasome activation.

机构信息

Department of Anesthesiology and Perioperative Medicine, Shanghai Key Laboratory of Anesthesiology and Brain Functional Modulation, Clinical Research Center for Anesthesiology and Perioperative Medicine, Translational Research Institute of Brain and Brain-Like Intelligence, Shanghai Fourth People's Hospital, School of Medicine, Tongji University, Shanghai, China.

出版信息

CNS Neurosci Ther. 2024 Oct;30(10):e70049. doi: 10.1111/cns.70049.

DOI:10.1111/cns.70049

PMID:39432407

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11493103/

Abstract

AIM

The aim of this study is to explore the key mechanisms of perioperative neurocognitive dysfunction (PND) after anesthesia/surgery (A/S) by screening hub genes.

METHODS

Transcriptome sequencing was conducted on hippocampal samples obtained from 18-month-old C57BL/6 mice assigned to control (Ctrl) and A/S groups. The functionality of differentially expressed genes (DEGs) was investigated using Metascape. Hub genes associated with changes between the two groups were screened by combining weighted gene coexpression network analysis within CytoHubba. Reverse transcription PCR and western blotting were used to validate changes in mRNA and protein expression, respectively. NLRP3 inflammasome activation was detected by western blotting and ELISA. Tauroursodeoxycholic acid (TUDCA), an inhibitor of endoplasmic reticulum (ER) stress, was administrated preoperatively to explore its effects on the occurrence of PND. Immunofluorescence analysis was performed to evaluate the activation of astrocytes and microglia in the hippocampus, and hippocampus-dependent learning and memory were assessed using behavioral experiments.

RESULTS

In total, 521 DEGs were detected between the control and A/S groups. These DEGs were significantly enriched in biological processes related to metabolic processes and their regulation. Four hub genes (Hspa5, Igf1r, Sfpq, and Xbp1) were identified. Animal experiments have shown that mice in the A/S group exhibited cognitive impairments accompanied by increased Hspa5 and Xbp1 expression, ER stress, and activation of NLRP3 inflammasome.

CONCLUSIONS

Inhibiting ER stress alleviated cognitive impairment in A/S mice; particularly, ER stress induced by A/S results in NLRP3 inflammasome activation and neuroinflammation. Moreover, the preoperative administration of TUDCA inhibited ER stress, NLRP3 inflammasome activation, and neuroinflammation.

摘要

目的

本研究旨在通过筛选枢纽基因来探讨麻醉/手术（A/S）后围手术期神经认知功能障碍（PND）的关键机制。

方法

对 18 月龄 C57BL/6 小鼠的海马样本进行转录组测序，将其分为对照组（Ctrl）和 A/S 组。使用 Metascape 研究差异表达基因（DEG）的功能。通过 CytoHubba 中的加权基因共表达网络分析，筛选与两组变化相关的枢纽基因。使用逆转录 PCR 和 Western blot 分别验证 mRNA 和蛋白表达的变化。通过 Western blot 和 ELISA 检测 NLRP3 炎性小体的激活。术前给予牛磺熊脱氧胆酸（TUDCA），一种内质网（ER）应激抑制剂，以探讨其对 PND 发生的影响。通过免疫荧光分析评估海马内星形胶质细胞和小胶质细胞的激活情况，并通过行为实验评估海马依赖性学习和记忆。

结果

共检测到对照组和 A/S 组之间 521 个 DEG。这些 DEG 显著富集于与代谢过程及其调控相关的生物学过程中。鉴定出 4 个枢纽基因（Hspa5、Igf1r、Sfpq 和 Xbp1）。动物实验表明，A/S 组小鼠表现出认知障碍，同时 Hspa5 和 Xbp1 表达增加、内质网应激和 NLRP3 炎性小体激活。