Sun Ye, Hu Yifan, Luo Shasha
Jiangnan University Wuxi School of Medicine, Wuxi No.2 People's Hospital (Jiangnan University Medical Center), Wuxi, 214000, China.
Department of Ophthalmology, Wuxi No.2 People's Hospital, Affiliated Wuxi Clinical College of Nantong University, Wuxi, 214000, China.
Med Microbiol Immunol. 2025 May 13;214(1):22. doi: 10.1007/s00430-025-00832-4.
The pathological basis of many visual disorders involves the abnormal viability and migration of retinal pigment epithelium (RPE) cells. Complement response disorder is a significant pathogenic factor causing some autoimmune and inflammation diseases. The complement activation product anaphylatoxin C5a signaling pathway may be associated with RPE cell dysfunction. This study aimed to analyze the molecular mechanisms by which C5a affects RPE cell viability and migration. Recombinant human complement component C5a protein stimulated RPE cells. Cell biological behavior, including cell viability, invasion, and migration were analyzed with Cell Counting Kit-8 and transwell methods. Bioinformatics analysis identified the differentially expressed genes (DEGs) involved in C5a-treated RPE cells based on RNA sequencing. SLC38A1 was knocked down or overexpressed by vector transfection to investigate its involvement in C5a-stimulated RPE cells. C5a promotes RPE cell viability and migration. C5a-induced DEGs are enriched in migration-associated pathways. C5a increased SLC38A1, and SLC38A1 knockdown or overexpression inhibited or promoted RPE cell viability and migration. Glutaminase inhibition abrogated the promoting effect of C5a and SLC38A1 on cell biological behaviors. METTL3-HNRNPC-mediated m6A modification mediated C5a-induced SLC38A1. C5a, METTL3, and SLC38A1 constituted a signaling axis in regulating cell biological behaviors of C5a-treated RPE cells. C5a promotes RPE cell viability and migration, and SLC38A1-mediated improved glutamine metabolism is the downstream signal pathway of the C5a complement pathway. The C5a complement system may target the SLC38A1 to promote RPE cell migration.
许多视觉障碍的病理基础涉及视网膜色素上皮（RPE）细胞的异常存活和迁移。补体反应紊乱是导致一些自身免疫性和炎症性疾病的重要致病因素。补体激活产物过敏毒素C5a信号通路可能与RPE细胞功能障碍有关。本研究旨在分析C5a影响RPE细胞存活和迁移的分子机制。重组人补体成分C5a蛋白刺激RPE细胞。采用细胞计数试剂盒-8和Transwell方法分析细胞生物学行为，包括细胞活力、侵袭和迁移。基于RNA测序的生物信息学分析确定了C5a处理的RPE细胞中差异表达的基因（DEGs）。通过载体转染敲低或过表达SLC38A1，以研究其在C5a刺激的RPE细胞中的作用。C5a促进RPE细胞的存活和迁移。C5a诱导的DEGs富集于迁移相关途径。C5a增加SLC38A1的表达，而SLC38A1的敲低或过表达抑制或促进RPE细胞的存活和迁移。谷氨酰胺酶抑制消除了C5a和SLC38A1对细胞生物学行为的促进作用。METTL3-HNRNPC介导的m6A修饰介导了C5a诱导的SLC38A1。C5a、METTL3和SLC38A1在调节C5a处理的RPE细胞的细胞生物学行为中构成了一个信号轴。C5a促进RPE细胞的存活和迁移，SLC38A1介导的谷氨酰胺代谢改善是C5a补体途径的下游信号通路。C5a补体系统可能靶向SLC38A1以促进RPE细胞迁移。
