Department of Ophthalmology, Yiwu Central Hospital, Yiwu, China.
Eur Rev Med Pharmacol Sci. 2019 Sep;23(18):7749-7756. doi: 10.26355/eurrev_201909_18984.
To elucidate the function of miRNA-138-5p in the early diabetic retinopathy (DR) and the potential mechanism.
DR model in rats was first established by streptozotocin (STZ) injection. MiRNA-138-5p expression in rat retinal tissues was determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Besides, its expression in retinal capillary endothelial cells (EC) and pericytes (RP) was also detected. Cell counting kit-8 (CCK-8) assay was performed to evaluate proliferative potentials of EC and RP cells. The target gene of miRNA-138-5p was predicted by bioinformatics and further confirmed by dual-luciferase reporter gene assay. Rescue experiments were carried out to verify whether the target gene could reverse the regulatory effect of miRNA-138-5p on the proliferation of EC and RP cells.
MiRNA-138-5p was lowly expressed in retinal tissues of DR rats, as well as in EC and RP cells. Overexpression of miRNA-138-5p suppressed the proliferative rate of EC and RP cells, and miRNA-138-5p knockdown obtained the opposite trends. NOVA1 was verified to be the target gene of miRNA-138-5p by dual-luciferase reporter gene assay and RIP assay, which was highly expressed in retinal tissues of DR rats, EC, and RP cells. MiRNA-138-5p knockdown markedly upregulated the mRNA and protein levels of NOVA1 in EC and RP cells. Of note, the inhibitory effect of miRNA-138-5p overexpression on proliferative potentials of EC and RP cells was reversed by NOVA1 overexpression. On the contrary, miRNA-138-5p knockdown accelerated their proliferative potentials and was further reversed by NOVA1 knockdown.
MiRNA-138-5p was lowly expressed in retinal tissues of DR rats, as well as in EC and RP cells. MiRNA-138-5p regulates the early DR by promoting cell proliferation via targeting NOVA1.
阐明 miRNA-138-5p 在早期糖尿病视网膜病变（DR）中的作用及其潜在机制。
首先通过链脲佐菌素（STZ）注射建立大鼠 DR 模型。通过定量实时聚合酶链反应（qRT-PCR）测定大鼠视网膜组织中 miRNA-138-5p 的表达。此外，还检测了其在视网膜毛细血管内皮细胞（EC）和周细胞（RP）中的表达。通过细胞计数试剂盒-8（CCK-8）测定评估 EC 和 RP 细胞的增殖潜力。通过生物信息学预测 miRNA-138-5p 的靶基因，并通过双荧光素酶报告基因检测进一步验证。进行挽救实验以验证靶基因是否可以逆转 miRNA-138-5p 对 EC 和 RP 细胞增殖的调节作用。
DR 大鼠视网膜组织及 EC 和 RP 细胞中 miRNA-138-5p 表达水平较低。过表达 miRNA-138-5p 抑制了 EC 和 RP 细胞的增殖率，而 miRNA-138-5p 敲低则获得了相反的趋势。双荧光素酶报告基因检测和 RIP 实验验证 NOVAl 是 miRNA-138-5p 的靶基因，并且在 DR 大鼠、EC 和 RP 细胞的视网膜组织中高表达。miRNA-138-5p 敲低显著上调了 EC 和 RP 细胞中 NOVAl 的 mRNA 和蛋白水平。值得注意的是，NOVA1 过表达逆转了 miRNA-138-5p 过表达对 EC 和 RP 细胞增殖潜能的抑制作用。相反，miRNA-138-5p 敲低加速了它们的增殖潜能，而 NOVAl 敲低进一步逆转了这一趋势。
DR 大鼠视网膜组织以及 EC 和 RP 细胞中 miRNA-138-5p 表达水平较低。miRNA-138-5p 通过靶向 NOVAl 促进细胞增殖来调节早期 DR。
