OBJECTIVES

This study used a quantitative bioinformatic analysis of public RNA sequencing databases to study key molecules mediating the occurrence of childhood constipation (CHC) and explore associated immune cell abnormalities and the role of natural killer (NK) cells.

METHODS

Gene expression profiling datasets, including CHC (GSE36701), were obtained. Immune genes were downloaded from the MSigDB database, and 3,907 immune-related genes were obtained. Differential analysis and weighted gene co-expression network analysis identified 12 hub genes related to CHC and immune genes in adipose tissue and blood samples from GSE36701. Gene ontology analysis was performed to determine key biologic processes. Stepwise and logistic regression analyses were performed to select specific genes for constructing a diagnostic model for CHC. The model was validated using GSE36701, and its diagnostic performance was assessed using the AUC value. The study recruited 20 CHC patients and 20 HV children. After blood collection, peripheral blood mononuclear cells (PBMCs) were extracted, and flow cytometry was used to detect the proportions of immune cells in the blood. qRT-PCR was employed to measure the expression of hub genes in NK cells.

RESULTS

A total of 12 hub genes were identified, among which the regulation of steroid metabolic processes and renal sodium excretion were closely associated with an increased expression level of CHC. Gene set enrichment analysis revealed that the core genes were associated with glycosphingolipid biosynthesis ganglio series, base excision repair, and ribosome characteristics. Immunoinfiltration analysis and experimental findings showed that the proportion of CD56 NK cells in patients with CHC was significantly lower compared to healthy children. The qRT-PCR results indicated that compared to the healthy volunteers group, the expression of AGTR1, FAM200B, NRSN2-AS1, PRAC1, SERTAD3, and TEF genes was decreased, while the expression of APANXA2-OT1, FMO9P, and LOC100506929 genes was increased in the CHC group.

CONCLUSION

This study identified TEF as a hub gene associated with the coexistence of CHC and immune cell abnormalities. The study highlights the important role of CD56 NK cells in the pathogenesis of CHC and provides possible targets for diagnosis and therapeutic intervention.