Chen Zhifeng, Shang Yulin, Ou Yanru, Zhou Li, Liu Ting, Gong Subo, Xiang Xudong, Peng Yating, Ouyang Ruoyun
Department of Pulmonary and Critical Care Medicine, The Second Xiangya Hospital, Central South University, Changsha, Hunan, 410011, People's Republic of China.
Research Unit of Respiratory Disease, Central South University, Changsha, Hunan, 410011, People's Republic of China.
Nat Sci Sleep. 2024 Dec 17;16:2063-2082. doi: 10.2147/NSS.S485952. eCollection 2024.
OSA can cause cognitive impairment (CI). The aim of this study was to investigate whether miR-20a-5p in exosomes derived from bEnd3 cells with IH mediates intercellular crosstalk and induces CI through hippocampal neuronal cell pyroptosis.
BEnd3-derived exosomes were isolated from the normal oxygen control group (NC-EXOS) and IH group (IH-EXOS). In vitro, exosomes were cocultured with HT22 cells. Meanwhile, in vivo, exosomes were injected into mice via the caudal vein. The spatial memory ability of mice was tested by MWM method to evaluate the effect of exosomes on the cognitive function of mice. Adults diagnosed with OSA underwent the MoCA and ESS tests to assess cognitive function and daytime sleepiness. Spearman's rank correlation analysis was used to evaluate the correlation between miR-20a-5p and candidate proteins and clinical parameters. Transfection using small interfering RNAs, miRNA mimics, and plasmids to evaluate the role of miR-20a-5p and its target genes. Dual luciferase reporter gene assay was used to confirm the binding of miR-20a-5p to its target gene.
IH could cause pyroptosis and inflammation in bEnd3 cells, and promote the expression of miR-20a-5p. Isolated IH-EXOS induced increased pyroptosis and activation of inflammatory response in vitro and in vivo, accompanied by increased expression of miR-20a-5p. In addition, IH-EXOS led to decreased learning and memory ability in mice. Interestingly, AHI was higher and MoCA scores were lower in severe OSA compared to healthy comparisons. In addition, miR-20a-5p and GSDMD were positively correlated with AHI but negatively correlated with MoCA in severe OSA. IH-induced exosomes were rich in miR-20a-5p, and these exosomes were found to deliver miR-20a-5p to HT22 cells, playing a key role in the induction of OSA-CI by directly targeting MFN2.
Exosome miR-20a-5p from IH-stimulated bEnd3 cells can promote OSA-CI by increasing HT22 cells pyroptosis through its target MFN2.
阻塞性睡眠呼吸暂停（OSA）可导致认知障碍（CI）。本研究旨在探讨间歇性低氧（IH）处理的bEnd3细胞来源的外泌体中的miR-20a-5p是否介导细胞间串扰并通过海马神经元细胞焦亡诱导CI。
从正常氧对照组（NC-EXOS）和IH组（IH-EXOS）中分离出bEnd3细胞来源的外泌体。在体外，将外泌体与HT22细胞共培养。同时，在体内，通过尾静脉将外泌体注射到小鼠体内。采用莫里斯水迷宫（MWM）法测试小鼠的空间记忆能力，以评估外泌体对小鼠认知功能的影响。对诊断为OSA的成年人进行蒙特利尔认知评估量表（MoCA）和 Epworth嗜睡量表（ESS）测试，以评估认知功能和日间嗜睡情况。采用Spearman等级相关分析评估miR-20a-5p与候选蛋白及临床参数之间的相关性。使用小干扰RNA、miRNA模拟物和质粒进行转染，以评估miR-20a-5p及其靶基因的作用。采用双荧光素酶报告基因测定法确认miR-20a-5p与其靶基因的结合。
IH可导致bEnd3细胞发生焦亡和炎症，并促进miR-20a-5p的表达。分离得到的IH-EXOS在体外和体内均可诱导焦亡增加和炎症反应激活，同时伴有miR-20a-5p表达增加。此外，IH-EXOS导致小鼠学习和记忆能力下降。有趣的是，与健康对照相比，重度OSA患者的呼吸暂停低通气指数（AHI）更高，MoCA评分更低。此外，在重度OSA中，miR-20a-5p和Gasdermin D（GSDMD）与AHI呈正相关，但与MoCA呈负相关。IH诱导的外泌体富含miR-20a-5p，并且发现这些外泌体将miR-20a-5p传递至HT22细胞，通过直接靶向线粒体融合蛋白2（MFN2）在OSA-CI的诱导中起关键作用。
来自IH刺激的bEnd3细胞的外泌体miR-20a-5p可通过其靶标MFN2增加HT22细胞焦亡来促进OSA-CI。
