Department of Neurology, General Hospital of Northern Theater Command, Shenyang, Liaoning 110013, P.R. China.
Department of Neurology, Jilin City Central Hospital, Jilin, Jilin 132011, P.R. China.
Mol Med Rep. 2022 Nov;26(5). doi: 10.3892/mmr.2022.12838. Epub 2022 Sep 2.
The purpose of the present study was to explore the functional role of microRNA (miR)‑363‑3p and related regulatory mechanisms in cerebral ischemia/reperfusion (I/R) injury. The neuronal cell line SH‑SY5Y was exposed to 4 h of oxygen and glucose deprivation (OGD), followed by 6, 12, 24 and 48 h of re‑oxygenation to mimic I/R injury . Cell viability, apoptosis and inflammation were assessed by CCK‑8, lactate dehydrogenase (LDH), flow cytometry and ELISA assays. The association between miR‑363‑3p and programmed cell death 6‑interacting protein (PDCD6IP) was further confirmed using luciferase reporter assay. Our data revealed that the expression level of miR‑363‑3p was significantly downregulated after OGD/R induction. Overexpression of miR‑363‑3p markedly suppressed OGD/R‑induced cell injury, as reflected by attenuated cell viability, reduced apoptosis, LDH activity and pro‑inflammatory cytokine levels. Mechanistically, PDCD6IP was confirmed as the target of miR‑363‑3p. Furthermore, PDCD6IP knockdown imitated, while overexpression reversed the effects of miR‑363‑3p overexpression on OGD/R‑induced cell injury. Collectively, miR‑363‑3p could attenuate OGD/R‑induced cell injury by alleviating apoptosis and inflammation, which may be mediated, at least in part, via inhibition of PDCD6IP.
本研究旨在探讨微小 RNA(miR)-363-3p 在脑缺血/再灌注(I/R)损伤中的功能作用及其相关调控机制。将神经元细胞系 SH-SY5Y 暴露于 4 h 的氧葡萄糖剥夺(OGD)中,随后再氧合 6、12、24 和 48 h 以模拟 I/R 损伤。通过 CCK-8、乳酸脱氢酶(LDH)、流式细胞术和 ELISA 测定评估细胞活力、细胞凋亡和炎症。通过荧光素酶报告实验进一步证实 miR-363-3p 与程序性细胞死亡 6 相互作用蛋白(PDCD6IP)之间的关联。我们的数据显示,miR-363-3p 的表达水平在 OGD/R 诱导后显著下调。miR-363-3p 的过表达显著抑制了 OGD/R 诱导的细胞损伤,表现为细胞活力降低、细胞凋亡减少、LDH 活性降低和促炎细胞因子水平降低。机制上,PDCD6IP 被确认为 miR-363-3p 的靶基因。此外,PDCD6IP 敲低模拟了 miR-363-3p 过表达对 OGD/R 诱导的细胞损伤的影响,而过表达则逆转了 miR-363-3p 过表达对 OGD/R 诱导的细胞损伤的影响。综上所述,miR-363-3p 可通过减轻细胞凋亡和炎症来减轻 OGD/R 诱导的细胞损伤,这至少部分是通过抑制 PDCD6IP 介导的。