摘要
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-145-5 p 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-145-5 p and FGF5 was verified by luciferase reporting experiment. The effects of mir-145-5 p 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-145-5 p 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-145-5 p promoted apoptosis and injury of neuronal cells by targeting FGF5. Conclusion This study provides a new therapeutic target for the treatment of ischemic stroke.
引文
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