miR-145-5p靶向FGF5对缺血/再灌注损伤诱导的神经细胞凋亡和氧化应激的调控作用

Regulatory effect of miR-145-5p targeting FGF5 on apoptosis and oxidative stress induced by ischemia/reperfusion injury

目的研究miR-145-5p对体外缺血/再灌注(I/R)损伤神经细胞模型的调控作用,并探讨其机制。方法运用氧-葡萄糖剥夺/复氧(OGD/R)建立脑缺血/再灌注损伤细胞模型。采用荧光素酶报告实验验证miR-145-5p与FGF5的靶向关系;RT-PCR、流式细胞术、Western blot等方法检测miR-145-5p对神经细胞HT22的凋亡和氧化应激损伤的影响。结果结果表明,与未进行OGD/R处理组(Normoxia组)相比,OGD/R处理6 h、12 h、24 h组凋亡率和氧化应激损伤明显增加。在OGD/R环境下,miR-145-5p inhibitor转染减轻HT22细胞凋亡和氧化应激损伤。然而,转染FGF5 siRNA逆转了这一效应。在OGD/R模拟的脑I/R损伤过程中,miR-145-5p通过靶向FGF5促进神经细胞凋亡和损伤。结论研究结果为缺血性脑卒中的治疗提供了一种新的治疗靶点。

Objective To investigate the regulatory effects of miR-145-5p on in vitro ischemia/reperfusion (I/R) injury of neurons and to explore their mechanisms. Methods The cell I/R injury model was established by oxygen-glucose deprivation/reoxygenation (OGD/R).The target relationship between mir-145-5p and FGF5 was verified by luciferase reporting experiment.The effects of mir-145-5p on apoptosis and oxidative stress in neuronal cell HT22 were detected by RT-PCR,flow cytometry and western blot. Results The results showed that the apoptosis rate and oxidative stress injury were significantly increased in the OGD/R treatment for 6 h,12 h,and 24 h compared with the group without OGD/R treatment (Normoxia).In OGD/R model,mir-145-5p inhibitor transfection reduced apoptosis and oxidative stress of HT22 cells.However,FGF5 siRNA transfected cells reversed this effect.In the process of brain I/R injury simulated by OGD/R,mir-145-5p promoted apoptosis and injury of neuronal cells by targeting FGF5. Conclusion This study provides a new therapeutic target for the treatment of ischemic stroke.