MicroRNA对老年性黄斑变性中血管紧张素Ⅱ1型受体的调控机制
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Regulation mechanism of microRNA on angiotensin II type 1 receptor in patients with age-related macular degeneration
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    摘要:

    目的:研究AMD患者的RPE细胞中微小RNA miR-410对血管紧张素受体Ⅱ的1型受体(AT1R)的调控效应。

    方法:实验分为AMD组、白内障组和正常组,运用生物信息学预测出AT1R是miR-410的靶基因,将正常的RPE细胞模拟AMD和白内障的微环境进行培养,检测其中miR-410的表达量,进一步将miR-410 mimics转染入细胞中,分别运用Q-PCR和Western blot的方法检测AT1R mRNA和蛋白的表达量,并通过双荧光素酶报告基因实验验证miR-410与AT1R的相互作用关系。

    结果:AMD组与白内障组和正常对照组相比,RPE细胞中的miR-410表达量显著降低(P=0.0006,0.0008),双荧光素酶报告基因实验表明,miR-410对AT1R具有明显的调控作用,且miR-410 mimics的下调效率大致为40%左右。细胞实验显示miR-410对AT1R的mRNA和蛋白表达的抑制率约为40%~50%。

    结论:AT1R是miR-410的靶基因,且在AMD的RPE细胞中提高miR-410的表达可以抑制AT1R的表达。

    Abstract:

    AIM: To study the effect of miR-410 on the regulation of angiotensin Ⅱ type 1 receptor(AT1R)in retinal pigment epithelium(RPE)cells of age-related macular degeneration(AMD)patients.

    METHODS: The experiment was divided into AMD patients, cataract patients and normal people group. AT1R was the target gene of miR-410 by bioinformatics, and the normal RPE cells were cultured in the simulated microenvironment of AMD and cataracts and the expression of miR-410 was detected. Then miR-410 mimics was transfected into cells, and the expression of mRNA and protein of AT1R were detected by Q-PCR and Western blot respectively. The relationship between miR-410 and AT1R was confirmed by the dual luciferase reporter assay.

    RESULTS: The miR-410 expression of in RPE cells with AMD was significantly reduced(P=0.0006, 0.0008)compared with cataract and normal controls. The miR-410 can regulate the function of AT1R by dual luciferase reporter gene experiment and the inhibition rate was about 40%. In addition, miR-410 inhibition rate was about 40%-50% to AT1R mRNA and protein expression by cell experiment.

    CONCLUSION: AT1R was a target gene of miR-410 in cell experiments, and it is demonstrated that increasing the expression of miR-410 in RPE cells with AMD can suppress the expression of AT1R.

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李霞. MicroRNA对老年性黄斑变性中血管紧张素Ⅱ1型受体的调控机制.国际眼科杂志, 2017,17(3):436-439.

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  • 收稿日期:2016-11-08
  • 最后修改日期:2017-02-17
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  • 在线发布日期: 2017-02-27
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