目的 探讨rno-miR-161调控egl-9家族缺氧诱导因子2(EGLN2)对血管性痴呆(vascular dementia,VD)大鼠额叶铁死亡的影响。 方法 24只雄性SD大鼠按随机原则分为模型组和假手术组,模型组结扎双侧颈总动脉,假手术组不结扎双侧颈总动脉。术后4周水迷宫检测行为学。RT-qPCR检测VD模型大鼠额叶rno-miR-161的变化,RT-qPCR和Western blotting检测EGLN2和谷胱甘肽过氧化物酶4(GPX4)的表达。生物信息学预测及双荧光素酶报告基因验证rno-miR-161与EGLN2是否存在靶向调控关系。RT-qPCR及Western blotting检测rno-miR-161-mimic和rno-miR-161-inhibitor对EGLN2表达的影响。siEGLN2转染PC12细胞,检测GPX4的表达。 结果 与假手术组相比,模型组大鼠额叶rno-miR-161表达下降、EGLN2表达上升、GPX4表达下降。rno-miR-161直接靶向EGLN2。rno-miR-161-mimic抑制EGLN2的表达,rno-miR-161inhibitor导致EGLN2表达升高。siEGLN2促进GPX4表达。 结论 VD大鼠额叶中rno-miR-161靶向EGLN2抑制铁死亡。
Abstract
Objective To explore the effect of rno-miR-161 on regulating egl-9 hypoxia-inducible factor 2(EGLN2) in frontal ferroptosis in vascular dementia (VD) rat. Methods Twenty-four adult male SD rats were randomly divided into a control group (sham), a model group (VD). The VD group was obtained by bilateral carotid artery ligation, while the sham group was not ligated with bilateral carotid artery. The Morris water maze test was applied to assess the symptoms of VD 4 weeks after surgery. RT-qPCR was used to detect the changes of rno-miR-161. RT-qPCR and Western blotting were used to detect the expression levels of EGLN2 and glutathione-dependent antioxidant enzyme glutathione peroxidase 4 (GPX4). The relationship between rno-miR-161 and EGLN2 was predicted by bioinformatics and luciferase reporter gene test. RT-qPCR and Western blotting were used to detect the effect of rno-miR-161-mimic and rno-miR-161-inhibitor on EGLN2 expression. The expression of EGLN2 was interfered by interference technology and the expression of GPX4 was detected by Western blotting and RT-qPCR. Results Compared with the control group, the expression of rno-miR-161 was down-regulated in the frontal lobe, the EGLN2 expression was increased while GPX4 was down-regulated. The EGLN2 was the direct target of rno-miR-161. rno-miR-161-minic inhibited the expression of EGLN2, rno-miR-161-inhibitor resulted in higher EGLN2 expression. siEGLN2 promoted GPX4. Conclusions rno-miR-161 inhibits ferroptosis by targeting EGLN2 expression in VD rat model frontal lobe.
关键词
血管性痴呆 /
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rno-miR-161 /
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EGLN2 /
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铁死亡 /
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GPX4
Key words
Vascular dementia /
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rno-miR-161 /
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EGLN2 /
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Ferroptosis /
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GPX4
中图分类号:
R361.3
R743.9 
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基金
国家自然科学基金(81673770,82174368)