Hierarchical cluster analysis in the study of the effect of cytokine expression patterns on endometrial repair and receptivity after hysteroscopic adhesiolysis

BACKGROUND: In a previous study, we reported that amnion promotes endometrial cell growth by regulating cytokines. In this study, hierarchical cluster analysis enabled the evaluation of cytokine expression changes after amnion treatment to be explored by cluster patterns. The role of IL1B on endometrial repair and receptivity was revealed. METHODS: A total of 30 patients were recruited in this clinical trial (NCT02496052) of hysteroscopic adhesiolysis. They were randomly allocated into an amnion grafts group (amnion group) and a control group. After hysteroscopic adhesiolysis, a Foley catheter covered with a sterilized freeze-dried amnion graft was inserted into the uterine cavity of the participants in the amnion group, whereas for the control group, a Foley catheter without amnion graft was inserted. After surgery, patient follow-up was done for a year. Uterine exudates were collected every day for seven days after surgery, and analyzed by enzyme-linked immunosorbent assays. Hierarchical cluster analysis was performed to compare expression patterns of each cytokine. Single-gene gene set enrichment analysis and differentially expressed genes enrichment analysis of IL1B were performed using NCBI GEO (N=151) to evaluate its potential mechanisms and impact on endometrial receptivity. RESULTS: Compared to the control group, cytokine expression patterns of the amnion group revealed significant stratifications, which were highly correlated with the expression levels of IL1B on the sixth to seventh day after surgery, improving the pregnant rate. Wilcoxon test revealed significantly low expression levels of IL1B in the reduced endometrial receptivity group compared to the normal group. Moreover, gene set enrichment analysis showed that lysosomes, cell cycle, and calcium signaling pathways were associated with the biological processes in which IL1B plays a role. Screening and enrichment analyses of differentially expressed genes further verified the mechanisms of action of IL1B on endometrial repair and receptivity recovery. CONCLUSIONS: Amnion promotes endometrial repair and receptivity by altering the expression levels and patterns of IL1B. Furthermore, by affecting lysosomal, cell cycle, and calcium signaling pathways, IL1B may be one of the factors involved in endometrial repair and receptivity recovery.