Department of Ophthalmology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, China; Hunan Clinical Research Center of Ophthalmic Diseases, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, China.
Department of Ophthalmology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, China; Hunan Clinical Research Center of Ophthalmic Diseases, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, China.
Int Immunopharmacol. 2024 Jun 15;134:112231. doi: 10.1016/j.intimp.2024.112231. Epub 2024 May 12.
The roles of immune cell infiltration and ferroptosis in the progression of proliferative diabetic retinopathy (PDR) remain unclear. To identify upregulated molecules associated with immune infiltration and ferroptosis in PDR, GSE60436 and GSE102485 datasets were downloaded from the Gene Expression Omnibus (GEO). Genes associated with immune cell infiltration were examined through Weighted Gene Co-expression Network Analysis (WGCNA) and CIBERSORT algorithm. Common differentially expressed genes (DEGs) were intersected with ferroptosis-associated and immune cell infiltration-related genes. Localization of cellular expression was confirmed by single-cell analysis of GSE165784 dataset. Findings were validated by qRT-PCR, ELISA, Western blotting, and immunofluorescence staining. As a result, the infiltration of M2 macrophages was significantly elevated in fibrovascular membrane samples from PDR patients than the retinas of control subjects. Analysis of DEGs, M2 macrophage-related genes and ferroptosis-related genes identified three hub intersecting genes, TP53, HMOX1 and PPARA. qRT-PCR showed that HMOX1 was significantly higher in the oxygen-induced retinopathy (OIR) mouse model retinas than in controls. Single-cell analysis confirmed that HMOX1 was located in M2 macrophages. ELISA and western blotting revealed elevated levels of HMOX1 in the vitreous humor of PDR patients and OIR retinas, and immunofluorescence staining showed that HMOX1 co-localized with M2 macrophages in the retinas of OIR mice. This study offers novel insights into the mechanisms associated with immune cell infiltration and ferroptosis in PDR. HMOX1 expression correlated with M2 macrophage infiltration and ferroptosis, which may play a crucial role in PDR pathogenesis.
免疫细胞浸润和铁死亡在增殖性糖尿病视网膜病变（PDR）进展中的作用尚不清楚。为了鉴定与 PDR 免疫浸润和铁死亡相关的上调分子，从基因表达综合数据库（GEO）下载了 GSE60436 和 GSE102485 数据集。通过加权基因共表达网络分析（WGCNA）和 CIBERSORT 算法检查与免疫细胞浸润相关的基因。将共同差异表达基因（DEGs）与铁死亡相关和免疫细胞浸润相关基因进行交集。通过 GSE165784 数据集的单细胞分析确认细胞表达的定位。通过 qRT-PCR、ELISA、Western blot 和免疫荧光染色验证发现。结果显示，与对照组相比，PDR 患者的纤维血管膜样本中 M2 巨噬细胞浸润明显增加。DEGs、M2 巨噬细胞相关基因和铁死亡相关基因分析确定了三个相交的关键基因，即 TP53、HMOX1 和 PPARA。qRT-PCR 显示，氧诱导的视网膜病变（OIR）小鼠模型视网膜中 HMOX1 明显高于对照组。单细胞分析证实 HMOX1 位于 M2 巨噬细胞中。ELISA 和 Western blot 显示 PDR 患者和 OIR 视网膜的玻璃体中 HMOX1 水平升高，免疫荧光染色显示 HMOX1 与 OIR 小鼠视网膜中的 M2 巨噬细胞共定位。这项研究为 PDR 中与免疫细胞浸润和铁死亡相关的机制提供了新的见解。HMOX1 的表达与 M2 巨噬细胞浸润和铁死亡相关，这可能在 PDR 的发病机制中起关键作用。
