Circular RNA circPRDX3 mediates neuronal survival apoptosis in ischemic stroke by targeting miR-641 and NPR3
Circular RNA circPRDX3 mediates neuronal survival apoptosis in ischemic stroke by targeting miR-641 and NPR3
复制标题
环状RNA circPRDX3通过靶向miR-641和NPR3介导缺血性中风中的神经元存活凋亡
DOI：
10.1016/j.brainres.2022.148114
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发表时间：
2022-10-12
期刊：
影响因子：
2.9
通讯作者：
Tu, Jianglong
中科院分区：
文献类型：
--
作者：
Chen, Weiping;Zhang, Yangbo;Tu, Jianglong
目的：circPRDX3是一种环状RNA（circRNA），目前还很少受到关注。本研究的目的是阐明circPRDX3在缺血性中风（IS）中的表达模式及其潜在网络。方法：分别使用氧糖剥夺/复氧（OGD/R）和大脑中动脉闭塞小鼠模型（MCAO）在N2a细胞或小鼠中生成IS模型。使用实时定量 PCR (qRT-PCR) 和蛋白质印迹测定 circPRDX3、miR-641、利钠肽受体 3 (NPR3) 和丝裂原激活蛋白激酶 (MAPK) 通路成员的表达水平。通过 CCK-8 测定评估细胞活力，并使用 TUNEL 染色和流式细胞术评估细胞凋亡。通过双荧光素酶和 RNA 免疫沉淀 (RIP) 测定验证分子间相互作用。通过氯化三苯基四唑 (TTC) 染色描绘梗塞区域，并使用美国国立卫生研究院卒中量表 (NIHSS) 测量神经功能水平。 结果：在 IS 样本以及 OGD/R 细胞或 MCAO 小鼠中，CircPRDX3 和 NPR3 显着下调，而 miR-641 显着上调。使用荧光素酶和 RIP 测定验证了 circPRDX3/miR-641/NPR3 机制。在 OGD/R 模型中，无论 MAPK 信号通路失活，circPRDX3 的过表达都会显着降低 miR-641 表达并增加 NPR3 表达，从而提高细胞存活率并降低细胞凋亡。此外，circPRDX3的过表达可降低MCAO后小鼠的梗塞体积并增强神经行为结果，而这些保护作用会因NPR3的消耗而显着消除。结论：总之，circPRDX3通过海绵化miR-641来抑制IS的发展，从而增加NPR3的表达并失活MAPK通路。这些结果可能有助于寻找 IS 的潜在治疗靶点。
Objective: circPRDX3 is a circular RNA (circRNA) that has received little attention yet. The purpose of this research is to elucidate circPRDX3 expression pattern and its underlying network in ischemic stroke (IS).Methods: Oxygen-glucose deprivation on/reoxygenation (OGD/R) and mice model of middle cerebral artery occlusion (MCAO) were used to generate IS model in N2a cells or mice, respectively. Expression levels of circPRDX3, miR-641, Natriuretic Peptide Receptor 3 (NPR3), and members of the mitogen-activated protein kinases (MAPK) pathway were determined using real-time quantitative PCR (qRT-PCR) and western blot. Cell viability was assessed by CCK-8 assay and apoptosis was evaluated using TUNEL staining and flow cytometry. Molecule-molecule interactions were verified by dual luciferase and RNA immunoprecipitation (RIP) assays. The infarcted area was depicted by Triphenyl tetrazolium chloride (TTC) staining and the level of neurological function was measured using National Institute of Health stroke scale (NIHSS).Results: CircPRDX3 and NPR3 were shown to be considerably downregulated in IS samples, as well as OGD/R cells or MCAO mice, while miR-641 was found to be significantly upregulated. A circPRDX3/miR-641/NPR3 mechinary was verified using luciferase and RIP assays. Overexpression of circPRDX3 dramatically reduced miR-641 expression and increased NPR3 expression, boosting cell survival and lowering apoptosis in an OGD/R model, either with inactivated MAPK signaling pathways. Moreover, overexpression of circPRDX3 lowered infarct volume and enhanced neurobehavioral outcomes in mice after MCAO, and these protective effects were dramatically abrogated by depletion of NPR3.Conclusion: Altogether, circPRDX3 inhibited the development of IS by sponging miR-641, hence increasing NPR3 expression and inactivating MAPK pathway. These results may aid in the search of potential therapy targets for IS.