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miR-325-3p Protects Neurons from Oxygen-Glucose Deprivation and Reoxygenation Injury via Inhibition of RIP3
Developmental Neuroscience ( IF 2.3 ) Pub Date : 2020-10-30 , DOI: 10.1159/000509108 Song Yi 1, 2 , Chuqin Zhang 3 , Na Li 4 , Yajing Fu 1 , Hongkun Li 1 , Jun Zhang 5
Developmental Neuroscience ( IF 2.3 ) Pub Date : 2020-10-30 , DOI: 10.1159/000509108 Song Yi 1, 2 , Chuqin Zhang 3 , Na Li 4 , Yajing Fu 1 , Hongkun Li 1 , Jun Zhang 5
Affiliation
Objective: Recent reports have corroborated that microRNAs (miRs) are related to the pathological changes of cerebral ischemia-reperfusion (CIR) induced injury. This work aimed to unearth the role and potential mechanism of miR-325-3p in regulating neuronal survival in CIR injury. Methods: To conduct this investigation, we established an in vitro model of CIR injury by subjecting neurons to oxygen-glucose deprivation and reoxygenation (OGD/R). Gain and loss of function of miR-325-3p and receptor-interacting serine-threonine kinase 3 (RIP3) in neurons were performed to observe its effect on cell apoptosis and the release of lactate dehydrogenase. The levels of miR-325-3p and RIP3 in neurons were detected by qRT-PCR. Western blot was employed to inspect the levels of caspase3, Bax, and Bcl-2, as well as p38 and JNK phosphorylation. The relationship between miR-325-3p and RIP3 was detected by TargetScan and validated by dual-luciferase reporter assay. Results: Firstly, miR-325-3p expression was obviously downregulated while RIP3 expression was upregulated in neurons following OGD/R treatment. Overexpressed miR-325-3p or downexpressed RIP3 ameliorated OGD/R-induced neuronal injury. Besides, RIP3 was a direct target mRNA of miR-325-3p. Additionally, Western blot revealed the mitogen-activated protein kinase (MAPK) pathway was involved in the regulation of miR-325-3p on OGD/R-induced neuronal injury. Furthermore, miR-325-3p was verified to hinder OGD/R-induced neuronal injury through downregulating RIP3. Conclusion: This study demonstrated that miR-325-3p targets RIP3 to inactivate the MAPK pathway, thereby protecting neurons against OGD/R-induced injury.
Dev Neurosci
中文翻译：
miR-325-3p 通过抑制 RIP3 保护神经元免受缺氧和复氧损伤
目的：近期报道证实微小RNA（miR）与脑缺血再灌注（CIR）损伤的病理变化有关。本工作旨在揭示 miR-325-3p 在调节 CIR 损伤中神经元存活中的作用和潜在机制。方法：为了进行这项研究，我们通过对神经元进行氧糖剥夺和复氧（OGD/R）来建立 CIR 损伤的体外模型。检测神经元中 miR-325-3p 和受体相互作用丝氨酸苏氨酸激酶 3 (RIP3) 功能的获得和丧失，观察其对细胞凋亡和乳酸脱氢酶释放的影响。采用qRT-PCR检测神经元中miR-325-3p和RIP3的水平。采用Western blot检测caspase3、Bax、Bcl-2的水平，以及p38、JNK的磷酸化水平。通过 TargetScan 检测 miR-325-3p 和 RIP3 之间的关系，并通过双荧光素酶报告基因测定进行验证。结果：首先，OGD/R处理后神经元中miR-325-3p表达明显下调，而RIP3表达上调。过表达的 miR-325-3p 或下调的 RIP3 可改善 OGD/R 诱导的神经元损伤。此外，RIP3是miR-325-3p的直接靶mRNA。此外，蛋白质印迹显示丝裂原激活蛋白激酶 (MAPK) 通路参与 miR-325-3p 对 OGD/R 诱导的神经元损伤的调节。此外，miR-325-3p 被证实可以通过下调 RIP3 来阻碍 OGD/R 诱导的神经元损伤。结论：本研究表明，miR-325-3p 靶向 RIP3 以使 MAPK 通路失活，从而保护神经元免受 OGD/R 诱导的损伤。
开发神经科学
更新日期：2020-10-30
Dev Neurosci
中文翻译：
miR-325-3p 通过抑制 RIP3 保护神经元免受缺氧和复氧损伤
目的：近期报道证实微小RNA（miR）与脑缺血再灌注（CIR）损伤的病理变化有关。本工作旨在揭示 miR-325-3p 在调节 CIR 损伤中神经元存活中的作用和潜在机制。方法：为了进行这项研究，我们通过对神经元进行氧糖剥夺和复氧（OGD/R）来建立 CIR 损伤的体外模型。检测神经元中 miR-325-3p 和受体相互作用丝氨酸苏氨酸激酶 3 (RIP3) 功能的获得和丧失，观察其对细胞凋亡和乳酸脱氢酶释放的影响。采用qRT-PCR检测神经元中miR-325-3p和RIP3的水平。采用Western blot检测caspase3、Bax、Bcl-2的水平，以及p38、JNK的磷酸化水平。通过 TargetScan 检测 miR-325-3p 和 RIP3 之间的关系，并通过双荧光素酶报告基因测定进行验证。结果：首先，OGD/R处理后神经元中miR-325-3p表达明显下调，而RIP3表达上调。过表达的 miR-325-3p 或下调的 RIP3 可改善 OGD/R 诱导的神经元损伤。此外，RIP3是miR-325-3p的直接靶mRNA。此外，蛋白质印迹显示丝裂原激活蛋白激酶 (MAPK) 通路参与 miR-325-3p 对 OGD/R 诱导的神经元损伤的调节。此外，miR-325-3p 被证实可以通过下调 RIP3 来阻碍 OGD/R 诱导的神经元损伤。结论：本研究表明，miR-325-3p 靶向 RIP3 以使 MAPK 通路失活，从而保护神经元免受 OGD/R 诱导的损伤。
开发神经科学
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