Bioinformatics analysis and experimental verification of ferroptosis in phospholipase A2 receptor associated membranous nephropathy
10.3760/cma.j.cn441217-20240827-00831
- VernacularTitle:磷脂酶A2受体相关膜性肾病中铁死亡的生物信息学分析与实验验证
- Author:
Manxia HUANG
1
;
Yuyang HE
;
Yangpu LI
;
Zhuoting LIU
;
Yanqing WANG
;
Youjia ZENG
Author Information
1. 广州中医药大学第四临床医学院，深圳 518033
- Publication Type:Journal Article
- Keywords:
Membranous nephropathy;
Phospholipase A2 receptor;
Ferroptosis;
Bioinformatics;
Genes
- From:
Chinese Journal of Nephrology
2025;41(2):81-89
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To identify and validate the key genes of ferroptosis in phospholipase A2 receptor (PLA2R) associated membranous nephropathy through bioinformatics analysis and in vitro experiments, and to explore the potential role of ferroptosis in PLA2R associated membranous nephropathy (PMN). Methods:The GSE115857 dataset obtained by retrieving the Gene Expression Omnibus (GEO) database and the ferroptosis-related genes obtained by retrieving the FerrDb database were intersected. The intersected genes were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. The key ferroptosis genes associated with PMN were identified by intersecting genes selected using support vector machines-recursive feature elimination and least absolute shrinkage and selection operator regression. The results were validate by real-time PCR, cell counting kit-8, Western blotting and immunofluorescence in human renal podocyte line AB 8/13 from both the control group and model group.Results:A total of 25 genes related to ferroptosis of PMN were obtained, and GO and KEGG analysis showed that these genes were mainly involved in cell ferroptosis metabolism. The key ferroptosis genes of PMN obtained by machine learning method were activating transcription factor 3 ( ATF3) and coiled coil domain containing 6 ( CCDC6). The results of in vitro experiments showed that the human renal podocyte line AB 8/13 in the model group was significantly deformed and retracted compared with the control group. The surface area density of foot processes was significantly reduced, and the podocyte cytoskeleton was allosteric. The morphology of F-actin was disordered and the expression of synaptopodin was decreased. The cell proliferation activity was significantly decreased ( P<0.05). The expression of PLA2R protein was increased ( P<0.05), and the expression of GPX4 protein was decreased ( P<0.01). The protein and mRNA levels of ATF3 and CCDC6 were significantly up-regulated (all P<0.05). Conclusions:Ferroptosis may be one of the key mechanisms in the occurrence and development of PMN. In vitro experiments show that ATF3 and CCDC6 are the key genes in the ferroptosis of PMN podocytes, which provides new insights and ideas for the pathogenesis of PMN.
