Pang Xueyi, Bai Siqiong, Feng Zhinan, Zhang Yumin, Hu Bojie, Zhang Yan
Tianjin Key Laboratory of Retinal Functions and Diseases, Tianjin Branch of National Clinical Research Center for Ocular Disease, Eye Institute and School of Optometry, Tianjin Medical University Eye Hospital, Tianjin, 300384, People's Republic of China.
Drug Des Devel Ther. 2025 Apr 17;19:2979-2999. doi: 10.2147/DDDT.S501318. eCollection 2025.
This study aimed to establish a high-fat diet (HFD)-induced rat model of type 2 diabetes mellitus (T2DM) and employed tandem mass tag (TMT) proteomics to search for novel interventional targets for nonproliferative diabetic retinopathy (NPDR).
Six-week-old male Sprague-Dawley rats were randomly divided into a T2DM group fed a HFD and a normal group (NOR group) fed normal chow. After 6 w, the T2DM group was confirmed to have impaired glucose tolerance and was intraperitoneally injected with a single small dose of streptozotocin (STZ, 30 mg/kg), and blood glucose levels were monitored. The HFD was maintained for another 6 w, and an Evans blue assay and a dark-adapted electroretinogram (ERG) were conducted. Rat retinas were collected for morphology analysis, TMT proteomics analysis, and Western blotting. The expression patterns of selected differentially expressed proteins (DEPs) were validated in rat retinas via Western blotting and in aqueous humor from NPDR patients via slot blotting.
After the 12-w HFD and STZ injection, the rats presented typical symptoms of T2DM. The retinas of T2DM rats presented pathological features of NPDR, including compromised scotopic ERGs, thinning of retinal layers, increased apoptosis and vascular leakage in the retina. Proteomic analysis identified DEPs and revealed profound dyslipidemia in T2DM rat retinas. The significant upregulation of the FABP3, TINAGL1, and COL4A3 proteins was validated in the retinas of the rats by Western blotting and in the aqueous humor of the NPDR patients by slot blotting.
In a rat model of HFD-induced T2DM that is consistent with the natural history and pathological features of NPDR, proteomics and bioinformatics analyses identified FABP3, TINAGL1, and COL4A3 as the 3 key upregulated proteins in retinas for the first time. These findings are supported by technical and clinical validations and provide novel targets for NPDR intervention.
本研究旨在建立高脂饮食（HFD）诱导的2型糖尿病（T2DM）大鼠模型，并采用串联质谱标签（TMT）蛋白质组学方法寻找非增殖性糖尿病视网膜病变（NPDR）的新型干预靶点。
将6周龄雄性Sprague-Dawley大鼠随机分为高脂饮食喂养的T2DM组和正常饮食喂养的正常组（NOR组）。6周后，确认T2DM组糖耐量受损，腹腔注射小剂量链脲佐菌素（STZ，30 mg/kg），并监测血糖水平。继续高脂饮食喂养6周，进行伊文思蓝试验和暗适应视网膜电图（ERG）检查。收集大鼠视网膜进行形态学分析、TMT蛋白质组学分析和蛋白质免疫印迹法检测。通过蛋白质免疫印迹法在大鼠视网膜中以及通过狭缝印迹法在NPDR患者的房水中验证所选差异表达蛋白（DEP）的表达模式。
经过12周的高脂饮食和STZ注射后，大鼠呈现出T2DM的典型症状。T2DM大鼠的视网膜呈现出NPDR的病理特征，包括暗视ERG受损、视网膜层变薄、视网膜细胞凋亡增加和血管渗漏。蛋白质组学分析鉴定出DEP，并揭示了T2DM大鼠视网膜中严重的血脂异常。通过蛋白质免疫印迹法在大鼠视网膜中以及通过狭缝印迹法在NPDR患者的房水中验证了FABP3、TINAGL1和COL4A3蛋白的显著上调。
在与NPDR自然病史和病理特征一致的高脂饮食诱导的T2DM大鼠模型中，蛋白质组学和生物信息学分析首次确定FABP3、TINAGL1和COL4A3为视网膜中3种关键上调蛋白。这些发现得到了技术和临床验证的支持，并为NPDR干预提供了新的靶点。
