Proteomic analysis of high glucose-induced apoptosis in peritoneal mesothelial cells

doi:10.3969/j.issn.1674-8115.2026.10.003

Abstract

Abstract:

Objective ·To investigate the biological mechanisms and molecular targets of high glucose-induced apoptosis in peritoneal mesothelial cells (PMCs) using proteomics. Methods ·The human peritoneal mesothelial cell line HMrSV5 was employed in this study. Cells were divided into a control group (untreated) and a high glucose group (treated with 138 mmol/L glucose). Cellular proteins were collected after 24 h of stimulation. Label-free quantitative proteomics was utilized to analyze changes in the protein expression profile of HMrSV5 cells under high glucose stimulation. Differentially expressed proteins (DEPs) were subjected to pathway enrichment analysis and protein-protein interaction (PPI) network construction. Cell proliferation viability was measured using the CCK-8 assay, and the apoptosis rate was detected by flow cytometry. Western blotting was performed to verify the expression and phosphorylation levels of the key protein checkpoint kinase 2 (CHEK2). Results ·A total of 7 366 proteins were identified. Principal component analysis (PCA) revealed significant differences between the control and glucose-treated groups. Compared with the control group, 417 DEPs were identified in the high glucose group, comprising 237 upregulated and 180 downregulated proteins. Subcellular localization analysis showed that upregulated DEPs were primarily distributed in the cytoplasm, nucleoplasm, and mitochondria, while downregulated DEPs were mainly located in the nucleoplasm and cytoplasm. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses indicated that DEPs were significantly enriched in cell cycle-related pathways, including the mitotic cell cycle, P53 signaling pathway, and cellular senescence. Gene Set Enrichment Analysis (GSEA) further suggested that the P53 signaling pathway was significantly positively enriched in the high glucose group, whereas the E2F target gene pathway and G₂/M checkpoint pathway were negatively enriched. PPI network analysis revealed that CHEK2 interacted with multiple cell cycle-related DEPs, suggesting it may be a potential key protein. CCK-8 and flow cytometry assays showed that high glucose stimulation reduced HMrSV5 cell viability and induced apoptosis. Western blotting results confirmed that high glucose stimulation significantly upregulated CHEK2 expression and increased its phosphorylation levels. Conclusion ·High glucose stimulation may promote apoptosis in PMCs by affecting cell cycle regulation, and CHEK2 may be a key regulatory protein.

Key words: peritoneal dialysis, proteomics, high glucose, apoptosis, cell cycle, checkpoint kinase 2 (CHEK2)

CLC Number:

R692.5

Li Ruolin, Li Zimeng, Song Ahui, Tao Ye, Li Chen, Liu Yingli. Proteomic analysis of high glucose-induced apoptosis in peritoneal mesothelial cells[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2026,(10): 1331-1338.

Figures/Tables 4

Fig 1 Proteomic alterations in high glucose-treated HMrSV5 cells

Fig 2 Functional enrichment analysis of DEPs in glucose-treated HMrSV5 cells

Fig 3 PPI network and expression patterns of cell cycle-related DEPs in glucose-treated HMrSV5 cells

Fig 4 Changes in cell viability, apoptosis level, and CHEK2 phosphorylation level after high-glucose stimulation of HMrSV5 cells

TrendMD

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