Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
The dataset used and/or analyzed in this study is available from the corresponding author on reasonable request.
References
Areloegbe SE, Peter MU, Oyeleke MB, Olaniyi KS (2022) Low-dose spironolactone ameliorates adipose tissue inflammation and apoptosis in letrozole-induced PCOS rat model. BMC Endocr Disord 22:224. https://doi.org/10.1186/s12902-022-01143-y
Babu A, Ramanathan G (2023) Multi-omics insights and therapeutic implications in polycystic ovary syndrome: a review. Funct Integr Genomics 23:130. https://doi.org/10.1007/s10142-023-01053-9
Cabrera-Ortega AA, Feinberg D, Liang Y, Rossa C Jr, Graves DT (2017) The role of forkhead box 1 (FOXO1) in the immune system: dendritic cells, T cells, B cells, and hematopoietic stem cells. Crit Rev Immunol 37:1–13. https://doi.org/10.1615/critrevimmunol.2017019636
Chen L, Gao B, Zhang Y et al (2019) PAR2 promotes M1 macrophage polarization and inflammation via FOXO1 pathway. J Cell Biochem 120:9799–9809. https://doi.org/10.1002/jcb.28260
Chen Y, Liu J, Zhang X et al (2022) lncRNA-GM targets Foxo1 to promote T cell-mediated autoimmunity. Sci Adv 8:eabn9181. https://doi.org/10.1126/sciadv.abn9181
Cirillo F, Catellani C, Sartori C et al (2019) CFTR and FOXO1 gene expression are reduced and high mobility group box 1 (HMGB1) is increased in the ovaries and serum of women with polycystic ovarian syndrome. Gynecol Endocrinol 35:842–846. https://doi.org/10.1080/09513590.2019.1599349
Joham AE, Norman RJ, Stener-Victorin E et al (2022) Polycystic ovary syndrome. Lancet Diabetes Endocrinol 10:668–680. https://doi.org/10.1016/s2213-8587(22)00163-2
Kunitomi C, Harada M, Kusamoto A et al (2021) Induction of aryl hydrocarbon receptor in granulosa cells by endoplasmic reticulum stress contributes to pathology of polycystic ovary syndrome. Mol Hum Reprod 27:gaab003. https://doi.org/10.1093/molehr/gaab003
Li Y, Yao N, Gao Y (2021) MiR-1224-5p attenuates polycystic ovary syndrome through inhibiting NOD-like receptor protein 3 inflammasome activation via targeting forkhead box O 1. Bioengineered 12:8555–8569. https://doi.org/10.1080/21655979.2021.1987125
Lima PDA, Nivet AL, Wang Q et al (2018) Polycystic ovary syndrome: possible involvement of androgen-induced, chemerin-mediated ovarian recruitment of monocytes/macrophages. Biol Reprod 99:838–852. https://doi.org/10.1093/biolre/ioy096
Luan YY, Zhang L, Peng YQ, Li YY, Liu RX, Yin CH (2022) Immune regulation in polycystic ovary syndrome. Clin Chim Acta 531:265–272. https://doi.org/10.1016/j.cca.2022.04.234
Miao ZL, Guo L, Wang YX et al (2012) The intervention effect of Rosiglitozone in ovarian fibrosis of PCOS rats. Biomed Environ Sci 25:46–52. https://doi.org/10.3967/0895-3988.2012.01.007
Qi X, Zhang B, Zhao Y et al (2017) Hyperhomocysteinemia promotes insulin resistance and adipose tissue inflammation in PCOS mice through modulating M2 macrophage polarization via estrogen suppression. Endocrinology 158:1181–1193. https://doi.org/10.1210/en.2017-00039
Regan SLP, Knight PG, Yovich JL, Leung Y, Arfuso F, Dharmarajan A (2018) Granulosa cell apoptosis in the ovarian follicle-a changing view. Front Endocrinol (lausanne) 9:61. https://doi.org/10.3389/fendo.2018.00061
Rostamtabar M, Esmaeilzadeh S, Tourani M et al (2021) Pathophysiological roles of chronic low-grade inflammation mediators in polycystic ovary syndrome. J Cell Physiol 236:824–838. https://doi.org/10.3390/ijms23020583
Sadeghi HM, Adeli I, Calina D et al (2022) Polycystic ovary syndrome: a comprehensive review of pathogenesis, management, and drug repurposing. Int J Mol Sci 23:583. https://doi.org/10.3390/ijms23020583
Shi F, LaPolt PS (2003) Relationship between FoxO1 protein levels and follicular development, atresia, and luteinization in the rat ovary. J Endocrinol 179:195–203. https://doi.org/10.1677/joe.0.1790195
Su Y, Gao J, Kaur P, Wang Z (2020) Neutrophils and macrophages as targets for development of nanotherapeutics in inflammatory diseases. Pharmaceutics 12:1222. https://doi.org/10.3390/pharmaceutics12121222
Wang D, Wang W, Liang Q et al (2018) DHEA-induced ovarian hyperfibrosis is mediated by TGF-β signaling pathway. J Ovarian Res 11:6. https://doi.org/10.1186/s13048-017-0375-7
Wang C, Ding C, Hua Z, Chen C, Yu J (2020) Cangfudaotan decoction alleviates insulin resistance and improves follicular development in rats with polycystic ovary syndrome via IGF-1-PI3K/Akt-Bax/Bcl-2 pathway. Mediators Inflamm 2020:8865647. https://doi.org/10.1155/2020/8865647
Wang Y, Yang Q, Wang H et al (2021) NAD+ deficiency and mitochondrial dysfunction in granulosa cells of women with polycystic ovary syndrome‡. Biol Reprod 105:371–380. https://doi.org/10.1186/s13048-017-0375-7
Xu R, Wang Z (2021) Involvement of transcription factor FoxO1 in the pathogenesis of polycystic ovary syndrome. Front Physiol 12:649295. https://doi.org/10.3389/fphys.2021.649295
Yang Y, Yang F, Yu X et al (2019) miR-16 inhibits NLRP3 inflammasome activation by directly targeting TLR4 in acute lung injury. Biomed Pharmacother 112:108664. https://doi.org/10.1016/j.biopha.2019.108664
Yang Z, Hong W, Zheng K et al (2022) Chitosan oligosaccharides alleviate H(2)O(2)-stimulated Granulosa cell damage via HIF-1α signaling pathway. Oxid Med Cell Longev 2022:4247042. https://doi.org/10.1155/2022/4247042
Zeng X, Xie YJ, Liu YT, Long SL, Mo ZC (2020) Polycystic ovarian syndrome: correlation between hyperandrogenism, insulin resistance and obesity. Clin Chim Acta 502:214–221. https://doi.org/10.1016/j.cca.2019.11.003
Zhang Z, Tian L, Jiang K (2019) Propofol attenuates inflammatory response and apoptosis to protect d-galactosamine/lipopolysaccharide induced acute liver injury via regulating TLR4/NF-κB/NLRP3 pathway. Int Immunopharmacol 77:105974. https://doi.org/10.1016/j.intimp.2019.105974
Zhang X, Wu H, Niu J et al (2023) A novel mitochondria-related gene signature in esophageal carcinoma: prognostic, immune, and therapeutic features. Funct Integr Genomics 23:109. https://doi.org/10.1007/s10142-023-01030-2
Zhao R, Jiang Y, Zhao S, Zhao H (2021) Multiomics analysis reveals molecular abnormalities in granulosa cells of women with polycystic ovary syndrome. Front Genet 12:648701. https://doi.org/10.3389/fgene.2021.648701
Zheng S, Chen N, Kang X, Hu Y, Shi S (2022) Irisin alleviates FFA induced β-cell insulin resistance and inflammatory response through activating PI3K/AKT/FOXO1 signaling pathway. Endocrine 75:740–751. https://doi.org/10.1007/s12020-021-02875-y
Zhou Y, Lan H, Dong Z et al (2022) Rhamnocitrin attenuates ovarian fibrosis in rats with letrozole-induced experimental polycystic ovary syndrome. Oxid Med Cell Longev 2022:5558599. https://doi.org/10.1155/2022/5558599
Zhou Z, Zhang P (2023) Formononetin ameliorates the LPS-induced inflammatory response and apoptosis of neuronal cells via NF-κB/NLRP3 signaling pathway. Funct Integr Genomics 23:321. https://doi.org/10.1007/s10142-023-01247-1
Funding
This work was financially supported by the Doctoral Supervisor Nursery Program of the Second Affiliated Hospital of Fujian Medical University (No: 2021GCC09 to X.C.) and Science and Technology Plan Project of Quanzhou (No: 2018Z110 to X.H.).
Ethics declarations
Ethics approval
The study was performed in accordance with the ethical standards as laid down in the Declaration of Helsinki and approved by the ethics committee of the Second Affiliated Hospital of Fujian Medical University.
Consent to participate
Not applicable.
Competing interests
The authors declare no competing interests.
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Huang, X., Luo, X., Huang, S. et al. Inhibition of FoxO1 alleviates polycystic ovarian syndrome by reducing inflammation and the immune response. Funct Integr Genomics 24, 6 (2024). https://doi.org/10.1007/s10142-024-01284-4
Received:
Revised:
Accepted:
Published:
DOI: https://doi.org/10.1007/s10142-024-01284-4