YTHDC1 aggravates high glucose-induced retinal vascular endothelial cell injury via m6A modification of CDK6.

IF 5.7 2区生物学 Q1 BIOLOGY

Biology Direct Pub Date : 2024-07-08 DOI:10.1186/s13062-024-00498-7

Qi Zhou, Min Tian, Yang Cao, Min Tang, Xiaohong Xiang, Lu Guo, Hongbin Lv

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Abstract

Objective: Retinal vascular endothelial cell (RVECs) injury is a major cause of morbidity and mortality among the patients with diabetes. RVECs dysfunction is the predominant pathological manifestation of vascular complication in diabetic retinopathy. N6-methyladenosine (m6A) serves as the most prevalent modification in eukaryotic mRNAs. However, the role of m6A RNA modification in RVECs dysfunction is still unclear.

Methods: RT-qPCR analysis and western blot were conducted to detect the change of m6A RNA modification in diabetic retinopathy. CCK-8 assay, transwell experiment, wound healing assay, tube formation experiment, m6A-IP-qPCR were performed to determine the role of YTHDC1 in RVECs. Retinal trypsin digestion test and H&E staining were used to evaluate histopathological changes.

Results: The levels of m6A RNA methylation were significantly up-regulated in HG-induced RVECs, which were caused by increased expression of YTHDC1. YTHDC1 regulated the viability, proliferation, migration and tube formation ability in vitro. YTHDC1 overexpression impaired RVECs function by repressing CDK6 expression, which was mediated by YTHDC1-dependent mRNA decay. Moreover, it showed sh-YTHDC1 inhibited CDK6 nuclear export. Sh-YTHDC1 promotes the mRNA degradation of CDK6 in the nucleus but does not affect the cytoplasmic CDK6 mRNA. In vivo experiments showed that overexpression of CDK6 reversed the protective effect of sh-YTHDC1 on STZ-induced retinal tissue damage.

Conclusion: YTHDC1-mediated m6A methylation regulates diabetes-induced RVECs dysfunction. YTHDC1-CDK6 signaling axis could be therapeutically targeted for treating DR.

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YTHDC1 通过 CDK6 的 m6A 修饰加重高糖诱导的视网膜血管内皮细胞损伤。

目的：视网膜血管内皮细胞（RVECs）损伤是糖尿病患者发病和死亡的主要原因：视网膜血管内皮细胞（RVECs）损伤是糖尿病患者发病和死亡的主要原因。视网膜血管内皮细胞功能障碍是糖尿病视网膜病变血管并发症的主要病理表现。N6-甲基腺苷（m6A）是真核生物 mRNA 中最常见的修饰。然而，m6A RNA修饰在RVECs功能障碍中的作用仍不清楚：方法：采用 RT-qPCR 分析和 Western 印迹检测糖尿病视网膜病变中 m6A RNA 修饰的变化。CCK-8实验、transwell实验、伤口愈合实验、管形成实验、m6A-IP-qPCR实验来确定YTHDC1在RVECs中的作用。用视网膜胰蛋白酶消化试验和 H&E 染色评估组织病理学变化：结果：HG 诱导的 RVECs 中 m6A RNA 甲基化水平明显上调，这是 YTHDC1 表达增加所致。YTHDC1调节RVECs在体外的活力、增殖、迁移和管形成能力。YTHDC1 的过表达通过抑制 CDK6 的表达损害了 RVECs 的功能，而 CDK6 的表达是由 YTHDC1 依赖性 mRNA 衰减介导的。此外，研究还发现sh-YTHDC1抑制了CDK6的核输出。sh-YTHDC1能促进CDK6在细胞核中的mRNA降解，但不影响细胞质中CDK6 mRNA的降解。体内实验表明，CDK6的过表达逆转了sh-YTHDC1对STZ诱导的视网膜组织损伤的保护作用：结论：YTHDC1介导的m6A甲基化调节糖尿病诱导的RVECs功能障碍。结论：YTHDC1 介导的 m6A 甲基化调控糖尿病诱导的 RVECs 功能障碍，YTHDC1-CDK6 信号轴可作为治疗 DR 的靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biology Direct 生物-生物学

CiteScore

6.40

自引率

10.90%

发文量

审稿时长

7 months

期刊介绍： Biology Direct serves the life science research community as an open access, peer-reviewed online journal, providing authors and readers with an alternative to the traditional model of peer review. Biology Direct considers original research articles, hypotheses, comments, discovery notes and reviews in subject areas currently identified as those most conducive to the open review approach, primarily those with a significant non-experimental component.