Sun Lihui, Liu Xuezheng, Zuo Zhongfu
Department of Anatomy, Histology and Embryology, Guangxi Medical University, Nanning, Guangxi Zhuang Autonomous Region 530021, P.R. China.
The Fifth Department of General Surgery, The Third Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning 121000, P.R. China.
Exp Ther Med. 2021 Dec;22(6):1477. doi: 10.3892/etm.2021.10912. Epub 2021 Oct 25.
The present study aimed to investigate the expression of microRNA (miRNA)-23a in blood and tear samples from diabetic retinopathy (DR) patients. Blood and tear samples were obtained from 33 patients with proliferative DR. Additionally, a rat model of DR was established. Reverse transcription-quantitative PCR was used to determine vascular endothelial growth factor (VEGF) mRNA and miRNA-23a expression levels, while ELISA and western blot analysis were performed to determine protein expression levels. Bioinformatics analysis and dual luciferase reporter assay were used to predict and validate the interaction between miRNA-23a and VEGF and cell proliferative ability was assessed with the MTT assay. In comparison to control patients VEGF mRNA and protein expression levels were significantly elevated in the blood and tear samples from patients with DR, while the expression level of miRNA-23a was significantly reduced. In blood and retinal tissues from a rat model of DR, the mRNA and protein expression levels of VEGF were significantly increased, while the miRNA-23a expression level was significantly decreased relative to controls. Dual luciferase reporter assay showed that miRNA-23a bound to the 3'-untranslated region (UTR) of VEGF. Moreover, over-expression of miRNA-23a significantly reduced the expression level of VEGF and the proliferative activity of human retinal microvascular endothelial cells. The elevated VEGF expression in the blood and tears of patients with DR may be related to the reduced miRNA-23a expression. miRNA-23a may regulate microvascular growth at the retina via VEGF and contribute to DR progression.
本研究旨在调查糖尿病视网膜病变(DR)患者血液和泪液样本中微小RNA(miRNA)-23a的表达情况。从33例增殖性DR患者中获取血液和泪液样本。此外,建立了DR大鼠模型。采用逆转录定量PCR测定血管内皮生长因子(VEGF)mRNA和miRNA-23a的表达水平,同时进行酶联免疫吸附测定(ELISA)和蛋白质免疫印迹分析以测定蛋白质表达水平。运用生物信息学分析和双荧光素酶报告基因检测来预测和验证miRNA-23a与VEGF之间的相互作用,并通过MTT法评估细胞增殖能力。与对照患者相比,DR患者血液和泪液样本中VEGF mRNA和蛋白质表达水平显著升高,而miRNA-23a的表达水平显著降低。在DR大鼠模型的血液和视网膜组织中,VEGF的mRNA和蛋白质表达水平显著升高,而相对于对照,miRNA-23a的表达水平显著降低。双荧光素酶报告基因检测表明,miRNA-23a与VEGF的3'非翻译区(UTR)结合。此外,miRNA-23a的过表达显著降低了VEGF的表达水平以及人视网膜微血管内皮细胞的增殖活性。DR患者血液和泪液中VEGF表达升高可能与miRNA-23a表达降低有关。miRNA-23a可能通过VEGF调节视网膜微血管生长,并促进DR的进展。