Chen Xinxin, Chen Xiaolong, Huang Shuai, Xian Yang, Han Ruining
China Medical University ophthalmology Shenyang China.
Curr Med Chem. 2025 Jun 18. doi: 10.2174/0109298673325802240813103408.
High glucose-induced angiogenesis is the main component in Proliferative Diabetic Retinopathy (PDR) development. In PDR, ischemia and hypoxia have been identified as key stimuli that promote pathological neoangiogenesis by increasing Vascular Endothelial Growth Factor A (VEGFA). Furthermore, it has been demonstrated that TRIM65 knockdown in tumor cells reduces VEGFA expression. Building on these findings, the present study aimed to study the role of TRIM protein members in proliferative diabetic retinopathy.
In comparison to the control group, TRIM65 expression was significantly increased in human retinal endothelial cells (HREC) after high glucose treatment. Moreover, FITC/PI staining, cell wound scratch assay, transwell assay, tube formation assay, and immunofluorescence staining of VEGFA and HIF-3α were carried out, which indicated that TRIM65 knockdown inhibited high glucose-induced HREC cell apoptosis and angiogenesis and decreased the expression of VEGFA and HIF-3α, both of which are potential targets of miR-29a-3p. MIR-29a-3p inhibitor significantly reduced the effects of TRIM65 knockdown on VEGFA and HIF-3α expression levels in cells. TRIM65 induced ubiquitination and degradation of TNRC6A, resulting in suppressed miR-29a-3p expression.
Furthermore, in vivo studies revealed that intravitreal injection of miR-29a-3p inhibited neoangiogenesis in mice with Oxygen-Induced Retinopathy (OIR). The retinal tissues of OIR mice showed higher TRIM65 mRNA expression and lower miR-29a-3p expression than those of control mice. Furthermore, the analysis showed a negative correlation between the expression of miR-29a-3p and TRIM65 in the retinal tissues of OIR mice.
In conclusion, this study demonstrated that the knockdown of TRIM65 inhibits neoangiogenesis in proliferative diabetic retinopathy by regulating miR-29a-3p.
高糖诱导的血管生成是增殖性糖尿病视网膜病变（PDR）发展的主要组成部分。在PDR中，缺血和缺氧已被确定为通过增加血管内皮生长因子A（VEGFA）促进病理性新生血管形成的关键刺激因素。此外，已经证明肿瘤细胞中TRIM65的敲低会降低VEGFA的表达。基于这些发现，本研究旨在探讨TRIM蛋白成员在增殖性糖尿病视网膜病变中的作用。
与对照组相比，高糖处理后人视网膜内皮细胞（HREC）中TRIM65的表达显著增加。此外，进行了FITC/PI染色、细胞划痕试验、Transwell试验、管形成试验以及VEGFA和HIF-3α的免疫荧光染色，结果表明TRIM65的敲低抑制了高糖诱导的HREC细胞凋亡和血管生成，并降低了VEGFA和HIF-3α的表达，这两者都是miR-29a-3p的潜在靶点。miR-29a-3p抑制剂显著降低了TRIM65敲低对细胞中VEGFA和HIF-3α表达水平的影响。TRIM65诱导TNRC6A的泛素化和降解，导致miR-29a-3p表达受到抑制。
此外，体内研究表明，玻璃体内注射miR-29a-3p可抑制氧诱导性视网膜病变（OIR）小鼠的新生血管形成。与对照小鼠相比，OIR小鼠的视网膜组织显示出更高的TRIM65 mRNA表达和更低的miR-29a-3p表达。此外，分析表明OIR小鼠视网膜组织中miR-29a-3p和TRIM65的表达呈负相关。
总之，本研究表明TRIM65的敲低通过调节miR-29a-3p抑制增殖性糖尿病视网膜病变中的新生血管形成。
