Poriferast-5-en-3β-ol improves pelvic inflammatory disease via reducing GADD45A-induced inflammation.

Abstract

BACKGROUND: Pelvic inflammatory disease (PID), often triggered by pathogens like Chlamydia, is a major cause of female infertility. This study investigated molecular mechanisms in PID and the therapeutic potential of DanZhiYin (DZY) and its components. METHODS: Bioinformatic analysis of dataset GSE110106 identified differentially expressed genes in PID, validated clinically. A Chlamydia infection model in OE-E6/E7 cells was used. siRNA knockdown and overexpression revealed GADD45A's role. Anti-inflammatory effects of DZY, Salvia miltiorrhiza, and Poriferast-5-en-3β-ol were tested via quantitative reverse transcription polymerase Chain Reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). Molecular dynamics and rescue experiments clarified the mechanism, and a PID mouse model assessed efficacy. RESULTS: MAPK signaling was upregulated in PID. GADD45A was identified as a key diagnostic marker, with an area under the curve (AUC) of 0.747. Chlamydia infection increased GADD45A and cytokines; its silencing reduced inflammation, while overexpression exacerbated it. DZY, Salvia miltiorrhiza, and Poriferast-5-en-3β-ol inhibited inflammation dose-dependently. Poriferast-5-en-3β-ol directly binds to GADD45A, thereby suppressing the production of IL-1β and TNF-α by inhibiting p38 MAPK and its phosphorylated form (p-p38). In vivo, all treatments reduced bacterial load and inflammation, with DZY being most effective. CONCLUSION: GADD45A is a central inflammatory regulator in PID. DZY and Poriferast-5-en-3β-ol attenuate inflammation by targeting GADD45A, providing new mechanistic and therapeutic insights for PID.