Temporal Metabolic Traits Induced by Androgen Excess Correlate with Cytokine Profiling in a Preclinical Mouse Model of PCOS

Background: Polycystic ovary syndrome (PCOS) is a multifaceted endocrine and metabolic disorder in reproductive-age women, characterized by androgen excess, ovulatory dysfunction, obesity, dyslipidemia, white adipose tissue (WAT) expansion, insulin resistance (IR), and also chronic low-grade inflammation. The inflammatory state in PCOS contributes to a vicious cycle in which excess androgens activate inflammatory pathways, and inflammation, in turn, further drives androgen excess and IR. Although both metabolic derangements and chronic inflammation are considered major contributors to PCOS pathophysiology, the onset and progression of these changes remain poorly understood. Therefore, using a well-established hyperandrogenemic mouse model of PCOS, this study aims to test the hypothesis that the non-aromatizable androgen dihydrotestosterone (DHT) induces a time-dependent progression of metabolic and inflammatory alterations. Methods: Three-week old female mice (C57BL/6N) were implanted with Silastic tubes filled with DHT (8 mg, s.c.) or vehicle (VEH) for 2, 4, 8 and 12 weeks (n=6/group). Weekly body weight (BW, gravimetry), body composition (EchoMRI), and final WAT depots (retroperitoneal fat (RPF), mesenteric fat (MF) and inguinal subcutaneous fat (iSCF)) mass (gravimetry) were assessed. Serum adiponectin, leptin and insulin were measured by ELISA, while total cholesterol (CHOL) was measured using a clinical chemistry analyzer. Circulating 23-plex (Bio-Plex Pro Mouse Cytokine, Group I) and 9-plex (Group II) cytokines were assessed by multiplex immunoassay. Correlation analysis was performed using Pearson correlation coefficient. Results: DHT-treated mice exhibited early metabolic alterations, including significant (p< 0.05) increases in BW (1.14-fold, 2 wks), lean mass (1.14-fold, 2 wks), RPF mass (3.28-fold, 4 wks), circulating insulin (2.31-fold, 2 wks), and CHOL (1.28-fold, 2 wks), alongside a 2-fold reduction in adiponectin (2 wks) compared to VEH. At later timepoints, DHT significantly increased fat mass (1.24-fold, 12 wks), MF mass (1.54-fold, 12 wks), iSCF mass (1.53-fold, 8 wks), and leptin (1.54-fold, 12 wks). Among the profiled circulating cytokines, the levels of several chemokines and macrophage activation-related factors (Eotaxin, MCP-1, MIP-1α, MIP-1β, and M-CSF) were significantly elevated during the early timepoints and negatively correlated with adiponectin in DHT-treated mice compared to VEH. In contrast, the levels of several pro-inflammatory (IL-1β, IL-5, IL-17A, KC, TNF-α), anti-inflammatory (IL-10, IL-13), pleotropic inflammatory (IL-2, IL-3, IL-6, IL-12p40) and growth factors (IL-3, PDGF-BB, GM-CSF, G-CSF) showed increases during the later timepoints, and positively correlated with fat mass and leptin. Conclusion and significance: These data show that hyperandrogenemia triggers early metabolic perturbations evidenced by increases in BW and visceral adiposity, reduced adiponectin, and elevated insulin, before broad inflammatory responses emerge. The early rise in chemokines and macrophage-related cytokines indicates that innate immune activation precedes and may drive the subsequent expansion of pro-, anti-, and pleiotropic inflammatory signals observed at later stages. Collectively, these findings reveal a temporal progression in which metabolic dysfunction and early immune activation interact to promote chronic inflammation. This underscores the need for early detection and intervention targeting metabolic and inflammatory risk factors to mitigate long-term PCOS dysregulations. Supported by NIH NIGMS grant P20GM121334 (L.L.Y.C., S.R. and D.G.R.), and AHA Predoctoral Fellowship 24PRE1200831 (J.B.). This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.