Department of Anesthesiology, China-Japan Union Hospital of Jilin University, Changchun 130033, Jilin, China.
Aging (Albany NY). 2022 Nov 30;14(24):9890-9907. doi: 10.18632/aging.204398.
Inhalation of sevoflurane can cause neuronal apoptosis, and cognitive disorders, inducing to the occurrence and progression of post operative cognitive dysfunction (POCD). This study aimed to explore the roles of sevoflurane-induced POCD-associated exosomes on HMC3 cells and its related mechanisms.
Exosomes were isolated from the plasma of sevoflurane-induced POCD or non-POCD patients, and were then sent for small RNA sequencing. Real-time quantitative PCR (RT-qPCR) was used to verify the sequencing results, and miR-584-5p was chosen for subsequent study. HMC3 cells were respectively transfected with POCD-derived exosomes and miR-584-5p mimics, and cell viability and apoptosis were measured. Dual-luciferase reporter gene assay was applied to confirm the target of miR-584-5p.
After sequencing, 301 differentially expressed miRNAs were identified, including 184 up-regulated miRNAs and 117 down-regulated miRNAs, and were significantly enriched in 3577 GO terms and 121 KEGG pathways. Due to the high level of miR-584-5p in sevoflurane-treated POCD-derived exosomes, HMC3 cells with miR-584-5p enrichment were successfully established. Compared with the control group, POCD-derived exosomes and miR-584-5p significantly inhibited viability and promoted apoptosis of HMC3 cells ( < 0.05). The IL-1β and TNF-α levels were significantly increased after POCD-derived exosomes and miR-584-5p mimics treatment compared to the control group ( < 0.05). Besides, POCD-derived exosomes and miR-584-5p mimics significantly down-regulated the expression levels of BDNF and p-TrkB, and up-regulated Caspase 3 and IL-1β. Finally, BDNF was confirmed to be the target of miR-584-5p.
Sevoflurane-induced POCD-associated exosomes delivered miR-584-5p may regulate the growth of HMC3 cells via targeting BDNF.
七氟醚吸入可引起神经元凋亡和认知障碍，导致术后认知功能障碍（POCD）的发生和发展。本研究旨在探讨七氟醚诱导的 POCD 相关外泌体对 HMC3 细胞的作用及其相关机制。
从七氟醚诱导的 POCD 或非 POCD 患者的血浆中分离出外泌体，进行小 RNA 测序。实时定量 PCR（RT-qPCR）用于验证测序结果，并选择 miR-584-5p 进行后续研究。分别用 POCD 衍生的外泌体和 miR-584-5p 模拟物转染 HMC3 细胞，测量细胞活力和凋亡。双荧光素酶报告基因检测用于确认 miR-584-5p 的靶基因。
测序后，鉴定出 301 个差异表达的 miRNA，包括 184 个上调 miRNA 和 117 个下调 miRNA，这些 miRNA 显著富集于 3577 个 GO 术语和 121 个 KEGG 通路中。由于七氟醚处理的 POCD 衍生外泌体中 miR-584-5p 的水平较高，因此成功建立了 miR-584-5p 富集的 HMC3 细胞。与对照组相比，POCD 衍生的外泌体和 miR-584-5p 显著抑制 HMC3 细胞的活力并促进其凋亡（<0.05）。与对照组相比，POCD 衍生的外泌体和 miR-584-5p 模拟物处理后，IL-1β 和 TNF-α 水平显著升高（<0.05）。此外，POCD 衍生的外泌体和 miR-584-5p 模拟物显著下调 BDNF 和 p-TrkB 的表达水平，并上调 Caspase 3 和 IL-1β。最后，BDNF 被证实是 miR-584-5p 的靶基因。
七氟醚诱导的 POCD 相关外泌体递送的 miR-584-5p 可能通过靶向 BDNF 调节 HMC3 细胞的生长。
