Wang Ning, Li Huibo, Sun Qinqin, Han Xuelian, Su Sheng
Eye Hospital, The First Affiliated Hospital, Harbin Medical University, Harbin, Heilongjiang, China.
Department of Hematology, The First Affiliated Hospital, Harbin Medical University, Harbin, Heilongjiang, China.
J Cell Mol Med. 2025 Mar;29(5):e70442. doi: 10.1111/jcmm.70442.
This study investigates the differences in ligand-receptor interactions between the communication network of vascular endothelial cells (ECs) and photoreceptor cells (PRCs)in diabetic retinopathy (DR) the mechanism was verified by animal experiments. The GSE209872 data set, including retinal specimens from five Sprague-Dawley rats induced by streptozotocin, was obtained from Gene Expression Omnibus. CM and EC data were extracted individually for reclustering, functional enrichment and trajectory analyses. Cell communication analysis was conducted to investigate the altered signals and significant ligand-receptor interactions. Moreover, novel ligand-receptor interactions were validated using immunofluorescence staining using 2, 4 and 8 weeks DR model; DR was treated with AAV-shANGPTL4, and retinal function was detected by Haematoxylin and eosin staining (HE), TUNEL and ELISA. The expression of ligand-receptor in DR Retina was detected by qPCR and immunohistochemistry. Nine cell types were determined in DR. Cellular communication results revealed four signalling pathways, including PTN, MK, ANGPTL and CXCL, that were significantly changed in DR. Furthermore, 3 ligand-receptor pairs (Ptn-Ncl, Mkd-Ncl and Angptl4-Sdc4) were obviously upregulated between ECs and PRCs, the expression of which was verified via immunofluorescence in the DR model. After treatment with AAV-shANGPTL4, the retinal thickness and average density of RGCs were decreased (p < 0.05). TUNEL staining showed that knocking down ANGPTL4 reduced the apoptosis of DR (p < 0.05), and VEGF and IGF-1 expression were downregulated (p < 0.01). The expression of ligand-receptors also decreased in the DR Model (p < 0.01). The vascular ECs and PRCs demonstrate significant heterogeneities in DR. ANGPTL4 was a decreased ligand-receptor expression and improved retinal function as a potential therapeutic target against DR.
本研究调查了糖尿病视网膜病变（DR）中血管内皮细胞（ECs）与光感受器细胞（PRCs）通信网络之间配体-受体相互作用的差异，并通过动物实验验证了其机制。从基因表达综合数据库（Gene Expression Omnibus）获取了GSE209872数据集，其中包括5只经链脲佐菌素诱导的斯普拉格-道利大鼠的视网膜标本。分别提取CM和EC数据进行再聚类、功能富集和轨迹分析。进行细胞通信分析以研究改变的信号和显著的配体-受体相互作用。此外，使用2、4和8周的DR模型通过免疫荧光染色验证了新的配体-受体相互作用；用腺相关病毒-shANGPTL4治疗DR，通过苏木精和伊红染色（HE）、TUNEL和ELISA检测视网膜功能。通过qPCR和免疫组织化学检测DR视网膜中配体-受体的表达。在DR中确定了9种细胞类型。细胞通信结果显示，DR中有4条信号通路，包括PTN、MK、ANGPTL和CXCL，发生了显著变化。此外，ECs和PRCs之间3对配体-受体（Ptn-Ncl、Mkd-Ncl和Angptl4-Sdc4）明显上调，其表达在DR模型中通过免疫荧光得到验证。用AAV-shANGPTL4治疗后，视网膜厚度和视网膜神经节细胞（RGCs）的平均密度降低（p<0.05）。TUNEL染色显示，敲低ANGPTL4可减少DR的细胞凋亡（p<0.05），血管内皮生长因子（VEGF）和胰岛素样生长因子-1（IGF-1）表达下调（p<0.01）。DR模型中配体-受体的表达也降低（p<0.01）。血管ECs和PRCs在DR中表现出显著的异质性。ANGPTL4作为一种潜在的抗DR治疗靶点，其配体-受体表达降低且视网膜功能得到改善。
