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[微小RNA-200b对高糖条件下视网膜内皮细胞功能的影响及其机制]

[Effect of MiR-200b on retinal endothelial cell function in high-glucose condition and the mechanism].

作者信息

Jiang Qun, Zhu Xiao-Hua, Liu Xin-Min, Liu Jian-Ming

机构信息

Department of Ophthalmology, Second Xiangya Hospital of Central South University, Changsha 410011, China. E-mail:

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2016 Apr;36(4):577-81.

Abstract

OBJECTIVE

To investigate the effect of MiR-200b on human retinal endothelial cells (hRECs) cultured in high glucose and explore the mechanism.

METHODS

hRECs cultured in high glucose or in normal media were examined for MiR-200b mRNA expression using real-time PCR. The effect of MiR-200b transfection on hREC proliferation in high-glucose culture was evaluated with MTT assay, and real-time PCR and Western blotting were performed to determine vascular endothelial growth factor (VEGF) and transforming growth factor β1 (TGFβ1) expression in the transfected cells.

RESULTS

The cells in high-glucose culture showed significantly decreased MiR-200b expression and active proliferation. Compared with those in normal control cells, VEGF and TGFβ1 mRNA and protein expressions increased markedly in cells cultured in high glucose (P<0.05). MiR-200b transfection of the cells caused significantly increased cellular expression of MiR-200b but decreased expression levels of VEGF and TGFβ1 mRNA and protein, and suppressed hREC proliferation in high glucose culture (P<0.05).

CONCLUSION

MiR-200b can regulate REC growth and proliferation by changing VEGF and TGFβ1 expressions and thus play a role in the pathogenesis and progression of diabetic retinopathy.

摘要

目的

研究MiR-200b对高糖培养的人视网膜内皮细胞(hRECs)的影响并探讨其机制。

方法

使用实时定量PCR检测在高糖或正常培养基中培养的hRECs的MiR-200b mRNA表达。采用MTT法评估MiR-200b转染对高糖培养中hREC增殖的影响,并通过实时定量PCR和蛋白质印迹法检测转染细胞中血管内皮生长因子(VEGF)和转化生长因子β1(TGFβ1)的表达。

结果

高糖培养的细胞中MiR-200b表达显著降低且增殖活跃。与正常对照细胞相比,高糖培养的细胞中VEGF和TGFβ1的mRNA及蛋白表达明显增加(P<0.05)。对细胞进行MiR-200b转染后,细胞中MiR-200b的表达显著增加,但VEGF和TGFβ1的mRNA及蛋白表达水平降低,并且在高糖培养中抑制了hREC的增殖(P<0.05)。

结论

MiR-200b可通过改变VEGF和TGFβ1的表达来调节视网膜内皮细胞的生长和增殖,从而在糖尿病视网膜病变的发病机制和进展中发挥作用。

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