Department of Ophthalmology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China.
National Clinical Research Center for Eye DiseasesShanghai Key Laboratory of Ocular Fundus DiseaseShanghai Engineering Center for Visual Science and Photo MedicineShanghai Engineering Center for Precise Diagnosis and Treatment of Eye Diseases, Shanghai, 200080, China.
Graefes Arch Clin Exp Ophthalmol. 2024 Jan;262(1):81-91. doi: 10.1007/s00417-023-06160-8. Epub 2023 Jun 27.
This work aimed to assess the possible role of TRIM25 in regulating hyperglycemia-induced inflammation, senescence, and oxidative stress in retinal microvascular endothelial cells, all of which exert critical roles in the pathological process of diabetic retinopathy.
The effects of TRIM25 were investigated using streptozotocin-induced diabetic mice, human primary retinal microvascular endothelial cells cultured in high glucose, and adenoviruses for TRIM25 knockdown and overexpression. TRIM25 expression was evaluated by western blot and immunofluorescence staining. Inflammatory cytokines were detected by western blot and quantitative real-time PCR. Cellular senescence level was assessed by detecting senescent marker p21 and senescence-associated-β-galactosidase activity. The oxidative stress state was accessed by detecting reactive oxygen species and mitochondrial superoxide dismutase.
TRIM25 expression is elevated in the endothelial cells of the retinal fibrovascular membrane from diabetic patients compared with that of the macular epiretinal membrane from non-diabetic patients. Moreover, we have also observed a significant increase in TRIM25 expression in diabetic mouse retina and retinal microvascular endothelial cells under hyperglycemia. TRIM25 knockdown suppressed hyperglycemia-induced inflammation, senescence, and oxidative stress in human primary retinal microvascular endothelial cells while TRIM25 overexpression further aggregates those injuries. Further investigation revealed that TRIM25 promoted the inflammatory responses mediated by the TNF-α/NF-κB pathway and TRIM25 knockdown improved cellular senescence by increasing SIRT3. However, TRIM25 knockdown alleviated the oxidative stress independent of both SIRT3 and mitochondrial biogenesis.
Our study proposed TRIM25 as a potential therapeutic target for the protection of microvascular function during the progression of diabetic retinopathy.
本研究旨在评估 TRIM25 在调节高血糖诱导的视网膜微血管内皮细胞炎症、衰老和氧化应激中的可能作用，这些作用在糖尿病性视网膜病变的病理过程中起着关键作用。
使用链脲佐菌素诱导的糖尿病小鼠、高糖培养的人原代视网膜微血管内皮细胞和用于 TRIM25 敲低和过表达的腺病毒来研究 TRIM25 的作用。通过 Western blot 和免疫荧光染色评估 TRIM25 的表达。通过 Western blot 和定量实时 PCR 检测炎症细胞因子。通过检测衰老标志物 p21 和衰老相关-β-半乳糖苷酶活性来评估细胞衰老水平。通过检测活性氧和线粒体超氧化物歧化酶来评估氧化应激状态。
与非糖尿病患者的黄斑视网膜内界膜相比，糖尿病患者的视网膜血管纤维膜内皮细胞中 TRIM25 的表达升高。此外，我们还观察到高血糖下糖尿病小鼠视网膜和视网膜微血管内皮细胞中 TRIM25 的表达显著增加。TRIM25 敲低抑制了人原代视网膜微血管内皮细胞中高血糖诱导的炎症、衰老和氧化应激，而 TRIM25 过表达进一步加剧了这些损伤。进一步的研究表明，TRIM25 促进了 TNF-α/NF-κB 通路介导的炎症反应，而 TRIM25 敲低通过增加 SIRT3 改善了细胞衰老。然而，TRIM25 敲低减轻了氧化应激，与 SIRT3 和线粒体生物发生均无关。
本研究提出 TRIM25 可能是糖尿病性视网膜病变进展过程中保护微血管功能的潜在治疗靶点。
