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首页> 外文期刊>Cell cycle >Circular RNA circ_HECTD1 regulates cell injury after cerebral infarction by miR-27a-3p/FSTL1 axis
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Circular RNA circ_HECTD1 regulates cell injury after cerebral infarction by miR-27a-3p/FSTL1 axis

机译:圆形RNA Circ_hectD1通过MiR-27A-3P / FSTL1轴调节脑梗塞后的细胞损伤

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摘要

Cerebral infarction is a common cerebrovascular disease caused by neural cell injury, with high mortality worldwide. Circular RNAs HECT domain E3 ubiquitin-protein ligase 1 (circ_HECTD1) has been reported to be related to the oxygen-glucose deprivation/reperfusion (OGD/R)-caused neuronal damage in cerebral ischemia. This study is designed to explore the role and mechanism of circ_HECTD1 in OGD/R-induced cell injury in cerebral ischemia. Circ_HECTD1, microRNA-27a-3p (miR-27a-3p), and Follistatin-like 1 (FSTL1) level were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The localization of circ_HECTD1 was analyzed by subcellular fractionation assay. Cell proliferative ability and apoptosis were assessed by 5-ethynyl-2 '-deoxyuridine (EdU), 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT), and flow cytometry assays. The protein levels of proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (Bcl-2), Bcl-2 related X protein (Bax), Cleaved poly-ADP-ribose polymerase (PARP), and FSTL1 were examined by western blot assay. The binding relationship between miR-27a-3p and circ_HECTD1 or FSTL1 was predicted by starbase 3.0 then verified by a dual-luciferase reporter assay. Circ_HECTD1 and FSTL1 were highly expressed, and miR-27a-3p was decreased in OGD/R-treated HT22 cells. Moreover, circ_HECTD1 knockdown could boost cell proliferative ability and repress apoptosis in OGD/R-triggered HT22 cells in vitro. Mechanical analysis discovered that circ_HECTD1 could regulate FSTL1 expression by sponging miR-27a-3p. Circ_HECTD1 deficiency could mitigate OGD/R-induced HT22 cell damage by modulating the miR-27a-3p/FSTL1 axis, providing a promising therapeutic target for cerebral infarction treatment.
机译:脑梗死是由神经细胞损伤引起的常见脑血管疾病,在世界范围内死亡率很高。据报道,环状RNA HECT结构域E3泛素蛋白连接酶1(circ_HECTD1)与脑缺血时氧葡萄糖剥夺/再灌注(OGD/R)引起的神经元损伤有关。本研究旨在探讨circ_HECTD1在OGD/R诱导的脑缺血细胞损伤中的作用及其机制。实时定量聚合酶链反应(RT-qPCR)检测Circ_Hect1、microRNA-27a-3p(miR-27a-3p)和卵泡抑素样1(FSTL1)水平。通过亚细胞分离分析circ_Hect1的定位。通过5-乙炔基-2'-脱氧尿苷(EdU)、3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴化铵(MTT)和流式细胞术检测细胞增殖能力和凋亡。免疫印迹法检测增殖细胞核抗原(PCNA)、B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、裂解聚ADP核糖聚合酶(PARP)和FSTL1的蛋白水平。miR-27a-3p与circ_Hect1或FSTL1之间的结合关系由starbase 3.0预测,然后通过双荧光素酶报告分析进行验证。在OGD/R处理的HT22细胞中,Circ_Hect1和FSTL1高表达,miR-27a-3p降低。此外,circ_HECTD1基因敲除可在体外提高OGD/R触发的HT22细胞的增殖能力并抑制凋亡。力学分析发现circ_HECTD1可以通过分泌miR-27a-3p来调节FSTL1的表达。Circ_HECTD1缺乏可通过调节miR-27a-3p/FSTL1轴减轻OGD/R诱导的HT22细胞损伤,为脑梗死治疗提供了一个有希望的治疗靶点。

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