| 张奕雯,凡子莲,李经伦,等.血脑屏障氧糖剥夺再灌体外模型小鼠脑内皮细胞 bEnd.3中微小 RNA-290b-3p的作用及机制研究[J].安徽医药,2021,25(2):345-348. |
| 血脑屏障氧糖剥夺再灌体外模型小鼠脑内皮细胞 bEnd.3中微小 RNA-290b-3p的作用及机制研究 |
| Study on the role and mechanism of miR-290b-3p in brain endothelium bEnd.3 of blood-brain barrier oxygen-glucose deprivation |
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| DOI：10.3969/j.issn.1009-6469.2021.02.032. |
| 中文关键词: 卒中 缺氧缺血，脑 再灌注损伤 氧糖剥夺 血脑屏障 紧密连接蛋白 Claudin-5 微小 RNA |
| 英文关键词: Stroke Hypoxia-Ischemia,brain Reperfusion injury OGD/R BBB Tight junction protein Claudin-5 microRNA |
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| 中文摘要: |
| 目的探讨氧糖剥夺再灌( OGD/R)条件下，小鼠脑内皮细胞 bEnd.3中微小 RNA-290b-3p(miR-290b-3p)在体外血脑屏障模型的作用与机制。方法培养小鼠脑内皮细胞 bEnd.3，建立 OGD/R模型与体外血脑屏障模型， Transwell法检测氧糖剥夺后体外血脑屏障模型通透性的变化；实时荧光定量聚合酶链式反应( real time-PCR，RT-PCR)检测 OGD/R后 miR-290b-3p表达水平； Targetscan网站预测 miR-290b-3p与紧密连接蛋白 5(Claudin-5)结合；荧光素酶报告实验验证 miR-290b-3p与 Claudin-5结合；转染 bEnd.3细胞 miR-290b-3p抑制剂，应用蛋白质印迹法( Western Blot)检测 Claudin-5的表达， Transwell法检测抑制 miR-290b-3p后氧糖剥夺后体外血脑屏障模型通透性变化。结果 OGD/R后体外血脑屏障破坏， 1、2、3、4、6h荧光素异硫氰酸酯( FITC)-葡聚糖扩散速率分别为( 0.04±0.00)、(0.04±0.01)、(0.05±0.01)、(0.07±0.01)、(0.12±0.01)pmol·mm-2·min-1通透性增加； miR-290b-3p在 OGD/R后 1、2、3、4、6h表达量分别为( 1.18±0.12)、(1.27±0.12)、(1.55±0.18)、(2.13±0.18)、(2.53±0.1，5)，表达增加； miR-290b-3p与 Claudin-5结合；抑制 miR-290b-3p可以缓解 OGD/R后 Claudin-5的降低，减轻血脑屏障的损伤。结论糖剥夺后 bEnd3细胞中 miR-290b-3p表达增加抑制 miR-290b-3p可通过调节 Claudin-5表达减轻氧糖剥夺模型中血脑屏障的氧损伤。 |
| 英文摘要: |
| Objective To explore the role of miR-290b-3p in the blood-brain barrier model in vitro under oxygen-glucose deprivation and reperfusion conditions.Methods Mouse brain endothelial cells bEnd.3 were cultured, oxygen glucose deprivation reperfusion model (OGD/R) and blood-brain barrier model in vitro were established, Transwell method was used to detect the permeability of blood-brain barrier model in vitro after oxygen glucose deprivation, and miR-290b-3p expression level after OGD/R was detected by real time-PCR. Targetscan algorithm was used to predict the interaction of miR-124 and Claudin-5; the binding of miR-290b-3p and Claudin-5 was detected by Luciferase; transfection of miR-290b-3p inhibitor in bEnd3 cells and detection of Claudin-5 expression by Western Blot and permeability of blood-brain barrier model in vitro.Results After OGD/R, the blood-brain barrier was destroyed, the diffusion rates of fluorescein isothiocyanate (FITC) -dextran were(0.04±0.00), (0.04±0.01), (0.05±0.01), (0.07±0.01), (0.12±0.01) pmol·mm-2· min-1.at 1, 2, 3, 4, and 6 h, respectively, and permeability increased. The expression of miR-290b-3p increased after OGD/R [1, 2, 3, 4, 6 h: (1.18±0.12), (1.27±0.12), (1.55±0.18), (2.13±0.18), (2.53±0.15)]; miR-290b-3p combined with Claudin-5; inhibition of miR-290b3p reversed decrease of Claudin-5 and alleviated the damage of the blood-brain barrier during OGD/R.Conclusion The expression of miR-290b-3p is increased induced by OGD/R in bEnd3, inhibition of miR-290b-3p attenuates blood-brain barrier damage in the oxygen-sugar deprivation model by modulating Claudin-5 expression. |
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