Department of Hyperbaric Oxygen, The Sixth Medical Center of PLA General Hospital, 6 Fucheng Road, Haidian District, Beijing, 100048, China.
Metab Brain Dis. 2023 Apr;38(4):1285-1296. doi: 10.1007/s11011-023-01178-7. Epub 2023 Feb 15.
Circular RNAs (circRNAs) are abundantly expressed in human central nervous system. Here, we explored the role of circ_Arhgap5 in cerebral ischemia/reperfusion (I/R)-induced nerve injury in PC12 cells and its associated mechanism. Cell proliferation ability was assessed by 5-Ethynyl-2'-deoxyuridine (Edu) assay and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Flow cytometry (FCM) was applied to assess cell apoptotic rate. Cell death was analyzed by lactate dehydrogenase (LDH) assay. Oxygen-glucose deprivation and reoxygenation (OGD/R) up-regulates the expression of circRNA Rho GTPase activating protein 5 (circ_Arhgap5; mmu_circ_0000377) in PC12 cells. OGD/R exposure inhibited the proliferation and induced the apoptosis of PC12 cells, and the silence of circ_Arhgap5 attenuated OGD/R-induced injury in PC12 cells. miR-29a-3p was identified as a target of circ_Arhgap5 in PC12 cells. Circ_Arhgap5 knockdown-mediated protective effects in OGD/R-induced PC12 cells were reversed by the interference of miR-29a-3p. miR-29a-3p interacted with the 3' untranslated region (3'UTR) of Rho-associated coiled-coil-containing protein kinase 1 (Rock1), and circ_Arhgap5 can positively regulate Rock1 expression by sponging miR-29a-3p in PC12 cells. miR-29a-3p overexpression protected PC12 cells against OGD/R-induced damage by down-regulating Rock1. In conclusion, circ_Arhgap5 silencing protected PC12 cells from OGD/R-induced injury through mediating miR-29a-3p/Rock1 axis.
环状 RNA（circRNA）在人类中枢神经系统中大量表达。在这里，我们探讨了 circ_Arhgap5 在 PC12 细胞脑缺血/再灌注（I/R）诱导的神经损伤中的作用及其相关机制。通过 5-乙炔基-2'-脱氧尿苷（Edu）检测和 3-(4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐（MTT）检测评估细胞增殖能力。流式细胞术（FCM）用于评估细胞凋亡率。通过乳酸脱氢酶（LDH）检测分析细胞死亡。氧葡萄糖剥夺和复氧（OGD/R）上调 PC12 细胞中环状 RNA Rho GTP 酶激活蛋白 5（circ_Arhgap5；mmu_circ_0000377）的表达。OGD/R 暴露抑制 PC12 细胞的增殖并诱导其凋亡，而 circ_Arhgap5 的沉默减轻了 OGD/R 诱导的 PC12 细胞损伤。miR-29a-3p 被鉴定为 PC12 细胞中 circ_Arhgap5 的靶标。circ_Arhgap5 敲低介导的 OGD/R 诱导的 PC12 细胞保护作用被 miR-29a-3p 的干扰所逆转。miR-29a-3p 与 Rho 相关卷曲螺旋蛋白激酶 1（Rock1）的 3'非翻译区（3'UTR）相互作用，circ_Arhgap5 可以通过在 PC12 细胞中海绵 miR-29a-3p 来正向调节 Rock1 表达。miR-29a-3p 过表达通过下调 Rock1 来保护 PC12 细胞免受 OGD/R 诱导的损伤。总之，circ_Arhgap5 沉默通过调节 miR-29a-3p/Rock1 轴来保护 PC12 细胞免受 OGD/R 诱导的损伤。
