Department of Anesthesiology, International Peace Maternity and Child Health Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Shanghai Key Laboratory of Embryo Original Diseases, Shanghai, China.
Brain Behav. 2023 May;13(5):e2936. doi: 10.1002/brb3.2936. Epub 2023 Mar 28.
Postoperative cognitive dysfunction (POCD) is a common complication of the central nervous system in elderly patients. The objective of this study was to investigate the role of methyltransferase 3 (METTL3) in the POCD progression.
The SH-SY5Y cells were treated with lipopolysaccharide (LPS) and exposed to sevoflurane to establish a POCD cell model. The cell viability and proliferation were assessed with MTT and EdU assays. Besides, the cell apoptosis was determined with TUNEL staining and flow cytometry. Additionally, the inflammatory factors were measured with ELISA. N6-methyladenosine (m6A) RNA Methylation Quantification Kit was used to detect the m6A levels. The relative expressions of methyltransferase 3 (METTL3) and Sex-determining region Y-box-2 (Sox2) was measured with RT-qPCR and western blot assays. RNA methylation immunoprecipitation-real-time quantitative PCR was performed to detect the RNA that was m6A modified.
After LPS treatment and sevoflurane exposure, the cell viability and proliferation were decreased and the cell apoptosis was elevated. The m6A and the METTL3 expression levels in the POCD cell model were declined. METTL3 overexpression promoted the cell growth and inhibited the cell apoptosis in the POCD cell model. Besides, the Sox2 levels were reduced in the POCD cell model. METTL3 silencing declined the m6A and mRNA levels of Sox2, while overexpression of METTL3 elevated it. The relationship between METTL3 and Sox2 was confirmed with double luciferase assay. Finally, Sox2 silencing neutralized the role of METTTL3 overexpression in the POCD cell model.
METTL3 relieved the injury of the SH-SY5Y cells induced by LPS treatment and sevoflurane exposure through regulating the m6A and mRNA levels of Sox2.
术后认知功能障碍（POCD）是老年患者中枢神经系统的常见并发症。本研究旨在探讨甲基转移酶 3（METTL3）在 POCD 进展中的作用。
用脂多糖（LPS）处理 SH-SY5Y 细胞，并使其暴露于七氟醚中，建立 POCD 细胞模型。用 MTT 和 EdU 测定细胞活力和增殖。此外，通过 TUNEL 染色和流式细胞术测定细胞凋亡。另外，通过 ELISA 测定炎症因子。用 N6-甲基腺苷（m6A）RNA 甲基化定量试剂盒检测 m6A 水平。用 RT-qPCR 和 Western blot 测定甲基转移酶 3（METTL3）和性别决定区 Y 框 2（Sox2）的相对表达。RNA 甲基化免疫沉淀-实时定量 PCR 检测 m6A 修饰的 RNA。
经 LPS 处理和七氟醚暴露后，细胞活力和增殖降低，细胞凋亡增加。POCD 细胞模型中的 m6A 和 METTL3 表达水平下降。METTL3 过表达促进 POCD 细胞模型中的细胞生长并抑制细胞凋亡。此外，POCD 细胞模型中的 Sox2 水平降低。METTL3 沉默降低了 Sox2 的 m6A 和 mRNA 水平，而过表达 METTL3 则升高了它。用双荧光素酶报告基因检测证实了 METTL3 和 Sox2 之间的关系。最后，Sox2 沉默中和了 METTL3 过表达在 POCD 细胞模型中的作用。
METTL3 通过调节 Sox2 的 m6A 和 mRNA 水平缓解 LPS 处理和七氟醚暴露对 SH-SY5Y 细胞的损伤。
