Shi Qiqin, Wang Qiangsheng, Mao Ke, Liu Zhuoran, Wang Ruobing
Department of Ophthalmology, Ningbo Hangzhou Bay Hospital, Ningbo, Zhejiang, 315000, China.
Department of Hematology, Ningbo Hangzhou Bay Hospital, Ningbo, Zhejiang, 315000, China.
Heliyon. 2024 Aug 5;10(15):e35663. doi: 10.1016/j.heliyon.2024.e35663. eCollection 2024 Aug 15.
Although anti-VEGF and retinal laser photocoagulation are two therapeutic modalities that have been used in the clinical treatment of diabetic retinopathy (DR), it is unknown how these modalities target vascular endothelial function in DR.
We first downloaded and analyzed the differential genes in two DR-related datasets, GSE60436 and GSE53257. The differential gene expression was then verified using RT-qPCR, and the most upregulated gene, NDUFB7, was selected for subsequent experiments. Subsequently, the role of NDUFB7 silencing and enforced expression on the proliferation and apoptosis of HRVECs was explored using CCK-8 assay, EDU proliferation assay and apoptotic TUNEL staining. In addition, the upstream potential miRNAs of NDUFB7 were predicted online using the Targetscan website. RT-qPCR, Western blotting (WB), and dual luciferase gene reporter assay were used to confirm the targeting connection between miR-2861 and NDUFB7. Finally, miR-2861 expression after high glucose (HG) treatment and its effect on proliferation and apoptosis of HRVECs under HG were investigated.
In this study, we first downloaded and analyzed the differential genes in two DR-related datasets, GSE60436 and GSE53257. We found that TUFM, PRELID1, MRPL32, NDUFB7, MRPL4, MRPL40, HSD17B10 and SLC25A13 were upregulated in DR, and RT-qPCR showed that NDUFB7 was most upregulated. Subsequent CCK-8 assay, EDU proliferation assay and TUNEL staining showed that up-picked NDUFB7 promotes proliferation and inhibits apoptosis of HRVECs. In addition, the upstream potential miRNAs of NDUFB7 were predicted online using the Targetscan website. RT-qPCR, Western blotting (WB), and dual luciferase gene reporter assay confirmed the targeting connection between miR-2861 and NDUFB7. Finally, it was observed that miR-2861 can inhibit the proliferation and promote the apoptosis of HRVECs by targeting NDUFB7.
Our findings showed that upregulated NDUFB7 in DR promotes proliferation and inhibits apoptosis of HRVECs, and miR-2861 can rescue the pathogenic effect of NDUFB7 upregulation by targeting NDUFB7.
尽管抗血管内皮生长因子（VEGF）和视网膜激光光凝是用于糖尿病视网膜病变（DR）临床治疗的两种治疗方式，但尚不清楚这些方式如何靶向DR中的血管内皮功能。
我们首先下载并分析了两个与DR相关的数据集中的差异基因，即GSE60436和GSE53257。然后使用逆转录定量聚合酶链反应（RT-qPCR）验证差异基因表达，并选择上调最明显的基因NDUFB7进行后续实验。随后，使用细胞计数试剂盒-8（CCK-8）检测、5-乙炔基-2'-脱氧尿苷（EDU）增殖检测和凋亡末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）染色，探讨NDUFB7沉默和过表达对人视网膜血管内皮细胞（HRVECs）增殖和凋亡的作用。此外，使用Targetscan网站在线预测NDUFB7的上游潜在微小RNA（miRNA）。采用RT-qPCR、蛋白质免疫印迹法（WB）和双荧光素酶基因报告检测法，证实miR-2861与NDUFB7之间的靶向关系。最后，研究高糖（HG）处理后miR-2861的表达及其对HG条件下HRVECs增殖和凋亡的影响。
在本研究中，我们首先下载并分析了两个与DR相关的数据集中的差异基因，即GSE60436和GSE53257。我们发现，TUFM、PRELID1、MRPL32、NDUFB7、MRPL4、MRPL40、HSD17B10和SLC25A13在DR中上调，RT-qPCR显示NDUFB7上调最为明显。随后的CCK-8检测、EDU增殖检测和TUNEL染色显示，筛选出的NDUFB7促进HRVECs增殖并抑制其凋亡。此外，使用Targetscan网站在线预测NDUFB7的上游潜在miRNA。RT-qPCR、WB和双荧光素酶基因报告检测法证实了miR-2861与NDUFB7之间的靶向关系。最后，观察到miR-2861可通过靶向NDUFB7抑制HRVECs增殖并促进其凋亡。
我们的研究结果表明，DR中上调的NDUFB7促进HRVECs增殖并抑制其凋亡，而miR-2861可通过靶向NDUFB7挽救NDUFB7上调的致病作用。
