Integrative bioinformatics and experimental approaches reveal NFKBIZ as a key regulator of inflammatory factors and chemokines in pelvic inflammatory disease progression

Purpose of the study

Pelvic inflammatory disease (PID) is a major reproductive health issue that can lead to complications such as infertility, chronic pelvic pain, and ectopic pregnancy. While PID has a significant clinical impact, its underlying immune mechanisms and inflammatory processes are not fully understood. This study aims to identify molecular changes, immune cell infiltration patterns, and key gene modules associated with PID progression, providing potential biomarkers for early diagnosis and targeted treatment.

Study design

We first analyzed the microarray dataset from GSE110106 to identify DEGs between PID patients and controls, and then performed functional enrichment analysis using GO and KEGG pathways. To investigate the immune status, we used seven algorithms to assess immune cell infiltration in PID samples. Additionally, WGCNA was employed to identify key gene modules associated with PID progression, while Least Absolute Shrinkage and Selection Operator (LASSO) regression was applied to identify potential signature genes. Experimental validation was performed using NIH-3T3 fibroblasts stimulated with cytokines to mimic PID conditions, followed by RT-PCR, ELISA and cell viability assays to assess the role of NFKBIZ in inflammation and cell damage.

Results

This study identified 7122 DEGs in PID patients, with 3317 up-regulated and 3805 down-regulated genes. Functional enrichment analysis highlighted key pathways such as NF-kappaB signaling and immune responses, while immune cell analysis revealed altered populations, particularly dendritic cells and T cell subsets. WGCNA identified a gene module most strongly correlated with PID, and machine learning methods, including LASSO regression, pinpointed NFKBIZ, KIAA0556, and SRGN as potential biomarkers. Validation experiments confirmed significant overexpression of NFKBIZ in a simulated PID environment, with knockdown of NFKBIZ reducing inflammatory mediator levels (IL-1β, TNF-α, MCP-1, MIP-2) and improving cell viability, suggesting NFKBIZ as a key player in PID inflammation and tissue damage.

Conclusion

Our findings suggest that NFKBIZ plays a crucial role in the progression of pelvic inflammatory disease by modulating inflammatory responses and promoting fibroblast apoptosis. Targeting NFKBIZ may offer a potential therapeutic approach for managing PID-related inflammation and tissue damage.