急性脑梗死发生过程中lncRNA TALNEC2的作用与机制

The role and mechanism of lncRNA TALNEC2 in the occurrence of acute cerebral infarction

目的分析长链非编码RNA TALNEC2在大脑中动脉阻塞(MCAO)模型和缺氧-葡萄糖剥夺(OGD)模型中的表达情况,并探讨其在急性脑梗死发生过程中的调节机制。方法建立MCAO小鼠模型和OGD细胞模型。采用qRT-PCR检测TALNEC2、miR-19a-3p和c-Jun氨基末端激酶(JNK)的表达。通过神经功能评分和TTC染色观察动物体内神经损伤情况。用Western blot、流式细胞仪和ELISA试验检测细胞凋亡和炎症损伤。通过荧光素酶活性分析miR-19a-3p与TALNEC2或JNK的相互作用。结果在MCAO模型和OGD模型中观察到TALNEC2上调。TALNEC2基因敲低可减轻小鼠MCAO模型的脑梗死、神经损伤、细胞凋亡和炎症损伤。荧光素酶活性分析显示miR-19a-3p与TALNEC2结合。miR-19a-3p过表达抑制了OGD处理的脑微血管内皮细胞(BMEC)的凋亡和炎症反应,TALNEC2的加入减弱了其抑制作用。JNK是miR-19a-3p的靶点,其上调减弱了miR-19a-3p对OGD处理的BMEC细胞的保护作用。此外,TALNEC2通过竞争性地吸附miR-19a-3p促进JNK的表达。结论TALNEC2的敲低通过调节miR-19a-3p/JNK减轻MCAO或OGD诱导的急性脑缺血损伤,为急性脑梗死的治疗提供了一种新的潜在策略。

Objective To analyze the expression of long non-coding RNA TALNEC2 in the middle cerebral artery occlusion(MCAO)model and oxygen-glucose deprivation(OGD)model,and to explore its regulatory mechanism in the process of acute cerebral infarction.Methods An in vivo mouse model of MCAO and an in vitro cell model of OGD were established to induce cerebral ischemic stroke condition.The expressions of TALNEC2,miR-19a-3p and c-jun-N-terminal kinase(JNK)were measured by qRT-PCR analysis.The neurological injury in vivo was investigated by neurological score and TTC staining.Cell apoptosis and inflammatory injury were analyzed by Western blot,flow cytometry and ELISA,respectively.The interaction between miR-19a-3p and TALNEC2 or JNK was explored by luciferase activity assays.Results Up regulation of TALNEC2 was observed in MCAO and OGD models.Knockdown of TALNEC2 attenuated the cerebral infarct,neurological injury,apoptosis and inflammatory injury in MCAO mice.Luciferase activity analysis showed that miR-19a-3p combined with TALNEC2.Overexpression of miR-19a-3p inhibited the apoptosis and inflammatory response in OGD-treated brain microvascular endothelial cell(BMEC),which was attenuated by TALNEC2.JNK was a target of miR-19a-3p and its restoration reversed the miR-19a-3p-mediated suppression of apoptosis and inflammation.Moreover,TALNEC2 promoted JNK expression by competitively sponging miR-19a-3p.Conclusion The knockdown of TALNEC2 attenuates MCAO-or OGD-induced acute cerebral ischemia injury by regulating miR-19a-3p/JNK,which provides a new potential strategy for the treatment of acute cerebral infarction.