Upregulation of miR-496 decreases cerebral ischemia/reperfusion injury by negatively regulating BCL2L14
Upregulation of miR-496 decreases cerebral ischemia/reperfusion injury by negatively regulating BCL2L14
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DOI：
10.1016/j.neulet.2018.12.039
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发表时间：
2019-03-23
影响因子：
2.5
通讯作者：
Huang, Fengzhen
中科院分区：
文献类型：
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作者：
Yao, Xiaoxi;Yao, Rui;Huang, Fengzhen
缺血性卒中后脑缺血再灌注（I/R）损伤严重损害神经功能，其分子机制尚不完全清楚。MicroRNA-496 （miR-496）已被报道在几种疾病中失调，但miR-496在脑I/R损伤中的功能尚不清楚。本研究采用大脑中动脉闭塞/再灌注法（MCAO/R）诱导大鼠大脑I/R损伤。然后采用Bederson和Longa评分系统评估神经功能缺损。采用2,3,5-三苯基四氯化铵（TTC）和末端脱氧核苷酸转移酶（UTP）缺口末端标记（TUNEL）染色对大鼠脑病理进行评价。在SH-SY5Y细胞中构建氧糖剥夺再灌注（OGD/R）体外I/R损伤模型。Real-time PCR检测miR-496的表达。利用生物信息学分析和双荧光素酶报告基因法确认miR-496的靶基因。CCK-8染色、EdU染色、流式细胞术和Hoechst 33258染色分别检测细胞活力、增殖和凋亡。结果显示，在大鼠脑I/R和OGD/R诱导的sh - sy5y细胞模型中，miR-496的表达下调。MiR-496过表达可减轻OGD/ r诱导的SH-SY5Y细胞损伤。此外，预测并验证了促凋亡因子bcl -2样蛋白14 （BCL2L14）是miR-496的直接靶点，并被miR-496抑制。此外，BCL2L14敲低表现出与miR-496过表达类似的效果，恢复BCL2L14表达逆转了miR-496对SH-SY5Y细胞的保护作用。总之，我们的研究结果表明miR-496可能通过抑制BCL2L14的表达来减轻脑I/R损伤。
Neurological functions were seriously impaired by cerebral ischemia-reperfusion (I/R) injury following ischemic stroke and its molecular mechanism is still not fully understood. MicroRNA-496 (miR-496) has been reported to be deregulated in several diseases but it still remains unknown about the function of miR-496 in cerebral I/R injury. Here, Middle cerebral artery occlusion/reperfusion (MCAO/R) was performed to induce cerebral I/R injury in rats. Then, neurological deficits were assessed by Bederson and Longa score system. The 2,3,5-triphenyltetrazolium chloride (TTC) and terminal deoxynucleotidyltransferase UTP nick end labeling (TUNEL) staining were used to evaluate the rat brain pathology. The oxygen-glucose deprivation and reperfusion (OGD/R) induced in vitro I/R injury was constructed in SH-SY5Y cells. The expression of miR-496 was determined using Real-time PCR. Bioinformatics analysis and dual luciferase reporter assay were utilized to confirm the target gene of miR-496. CCK-8, EdU staining, flow cytometry and Hoechst 33258 staining were respectively utilized to measure cell viability, proliferation and apoptosis. The results showed the expression of miR-496 was found to be down-regulated by cerebral I/R in rats and OGD/R-inducedSH-SY5Y cell model. MiR-496 overexpression alleviated the OGD/R-induced injury in SH-SY5Y cells. In addition, pro-apoptosis factor Bcl-2-like protein 14 (BCL2L14) was predicted and verified as the direct target of miR-496 and suppressed by miR-496. Furthermore, BCL2L14 knockdown exhibited similar effects similar to those of miR-496 overexpression, and the restored BCL2L14expression reversed the protective effects of miR-496 on SH-SY5Y cells. In conclusions, our results suggest that miR-496 alleviates cerebral I/R injury possibly via inhibiting BCL2L14 expression.