MiRNA-138-5p protects the early diabetic retinopathy by regulating NOVA1
MiRNA-138-5p protects the early diabetic retinopathy by regulating NOVA1
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DOI：
10.26355/eurrev_201909_18984
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发表时间：
2019-01-01
影响因子：
3.3
通讯作者：
Cao, J.
中科院分区：
文献类型：
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作者：
Bao, X-Y;Cao, J.
目的：为探讨miRNA-138- 5 p在早期糖尿病视网膜病变（diabetic retinopathy，DR）中的作用及其可能机制，采用链脲佐菌素（streptozotocin，STZ）建立大鼠DR模型。通过定量真实的时间-聚合酶链反应（qRT-PCR）测定大鼠视网膜组织中miRNA-138- 5 p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。采用CCK-8法检测EC和RP细胞的增殖能力。通过生物信息学方法预测了miRNA-138- 5 p的靶基因，并通过双荧光素酶报告基因分析进一步证实了miRNA-138- 5 p的靶基因。结果：miRNA-138 - 5 p在DR大鼠视网膜组织中低表达，在EC和RP细胞中也低表达。miRNA-138- 5 p过表达可抑制EC和RP细胞的增殖。而miRNA-138- 5 p敲低则呈现相反的趋势。通过双荧光素酶报告基因检测和RIP检测，证实NOVA 1是miRNA-138- 5 p的靶基因，在DR大鼠视网膜组织、EC和RP细胞中高表达。miRNA-138- 5 p敲低显著上调EC和RP细胞中NOVA 1的mRNA和蛋白水平。值得注意的是，miRNA-138- 5 p过表达对EC和RP细胞增殖潜力的抑制作用被NOVA 1过表达逆转。结论：miRNA-138 - 5 p在DR大鼠视网膜组织中低表达，在EC和RP细胞中也低表达。miRNA-138- 5 p通过靶向NOVA 1促进细胞增殖来调节早期DR。
OBJECTIVE: To elucidate the function of miRNA-138-5p in the early diabetic retinopathy (DR) and the potential mechanism.MATERIALS AND METHODS: 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.RESULTS: 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.CONCLUSIONS: 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.