The aim of this study was to explore the role of forkhead box transcription Factor O1 (FoxO1) in chronic inflammation in polycystic ovary syndrome (PCOS). A PCOS rat model was constructed as an in vivo model by letrozole induction, and granulosa cells (GCs) from PCOS rats were isolated and cultured as an in vitro cellular model. FoxO1 was knocked down by shRNA and siRNA in the PCOS rat model and GCs model, respectively. H&E staining was conducted to evaluate the effect of FoxO1 inhibition on ovarian pathology and dysfunction in PCOS rats. The levels of inflammatory cytokines in the ovaries and uterus of PCOS rats and in GCs were assessed by ELISA. Flow cytometry was used to evaluate the changes in the contents of neutrophils and macrophages in the peripheral blood and spleen of PCOS rats. CCK-8 assays and Annexin V-FITC/PI staining were performed to evaluate the proliferation and apoptosis of GCs. The expression of genes and proteins related to the TLR4/NF-κB/NLRP3 pathway in GCs was determined by RT-qPCR and Western blotting. The results indicated that FoxO1 was highly expressed in PCOS rat model. Inhibition of FoxO1 significantly mitigated the pathological changes and dysfunction in the ovaries of PCOS rats while also suppressing inflammation and fibrosis in the ovaries and uterus. Moreover, knocking down FoxO1 facilitated the restoration of the normal ratio of neutrophils and macrophages in the peripheral blood and spleen of PCOS rats and promoted M2 polarization of macrophages. Additionally, inhibition of FoxO1 promoted the proliferation of GCs and inhibited the inflammatory response in GCs. Furthermore, FoxO1 knockdown inhibited the activation of the NF-κB pathway and the formation of the NLRP3 inflammasome in GCs. In conclusion, inhibition of FoxO1 can alleviate PCOS by inhibiting the TLR4/NF-κB/NLRP3 pathway to reduce inflammation and the immune response.

本研究的目的是探讨叉头盒转录因子 O1 (FoxO1) 在多囊卵巢综合征 (PCOS) 慢性炎症中的作用。通过来曲唑诱导构建PCOS大鼠模型作为体内模型，并分离培养PCOS大鼠的颗粒细胞（GC）作为体外细胞模型。在 PCOS 大鼠模型和 GC 模型中，FoxO1 分别被 shRNA 和 siRNA 敲低。通过 H&E 染色评估 FoxO1 抑制对 PCOS 大鼠卵巢病理和功能障碍的影响。通过 ELISA 评估 PCOS 大鼠卵巢和子宫以及 GC 中炎症细胞因子的水平。采用流式细胞术评价PCOS大鼠外周血及脾脏中性粒细胞、巨噬细胞含量的变化。进行CCK-8测定和Annexin V-FITC/PI染色来评估GC的增殖和凋亡。通过RT-qPCR和Western blotting检测GC中TLR4/NF-κB/NLRP3通路相关基因和蛋白的表达。结果表明FoxO1在PCOS大鼠模型中高表达。抑制FoxO1可显着减轻PCOS大鼠卵巢的病理变化和功能障碍，同时抑制卵巢和子宫的炎症和纤维化。此外，敲低FoxO1有利于PCOS大鼠外周血和脾脏中中性粒细胞和巨噬细胞的正常比例恢复，促进巨噬细胞M2极化。此外，抑制FoxO1可促进GC的增殖并抑制GC的炎症反应。此外，FoxO1 敲低抑制了 GC 中 NF-κB 通路的激活和 NLRP3 炎症小体的形成。总之，抑制FoxO1可以通过抑制TLR4/NF-κB/NLRP3通路来减轻炎症和免疫反应，从而缓解PCOS。