To investigate the potential miRNA-mRNA network co-expressed in polycystic ovary syndrome (PCOS) and diabetes, and explore the molecular mechanism of traditional acupuncture treatment of PCOS.Patients with PCOS and diabetes who had undergone acupuncture treatment from January 2019 to June 2020 were recruited in this study. The potential miRNA-mRNA network co-expressed in PCOS and diabetes was obtained through bioinformatics analysis. The expression levels of candidate gen es were determined using quantitative qRT-PCR to study the effectiveness of acupuncture approach. Further, the mechanism of action of acupuncture method was determined using luciferase assay.A total of 44 patients were included in this study. The miRNA-mRNA network for PCOS was then constructed based on the results of the bioinformatics analysis. Acupuncture treatment could significantly down-regulate miR-32-3p levels and up-regulate expression of PLA2G4A. Luciferase experiments showed that miR-32-3p could affect glucose metabolism in PCOS patients through down-regulating PLA2G4A expression. Functional and pathway enrichment analysis further suported this finding.MiR-32-3p regulates PLA2G4A protein expression, which is vital in the pathogenesis of PCOS and diabetes. Further, this research proved that the potential mechanism of traditional acupuncture treatment may be the downregulation of miR-32-3p, thus inhibiting PCOS and diabetes progression.AJTR Copyright © 2021.