{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:53:26Z","timestamp":1785405206891,"version":"3.56.0"},"reference-count":45,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T00:00:00Z","timestamp":1779667200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Naval Medical University Continuation Fund","award":["2024YX005"],"award-info":[{"award-number":["2024YX005"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["82071598"],"award-info":[{"award-number":["82071598"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["82071598"],"award-info":[{"award-number":["82071598"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCM"],"abstract":"<jats:p>Objectives: Diminished ovarian reserve (DOR) significantly compromises in vitro fertilization (IVF) success. Although systemic markers such as anti-M\u00fcllerian hormone (AMH) serve as valuable clinical indicators of the ovarian reserve, they lack the sensitivity to reflect the qualitative deterioration of the follicular microenvironment. Therefore, in this study, we aimed to characterize the inflammatory proteome of follicular fluid (FF) to establish a high-performance auxiliary diagnostic model for DOR. Methods: Utilizing the ultra-sensitive Olink proximity extension assay, we quantified 92 inflammation-related proteins in the FF of 88 participants (67 with DOR and 21 normal controls). Differentially expressed proteins (DEPs) were identified, and their relationships with key clinical indices were evaluated. A robust predictive signature was refined through integrated Least Absolute Shrinkage and Selection Operator (LASSO) regression and Random Forest algorithms, with diagnostic performance assessed via 10-fold cross-validation. Results: Thirty-five DEPs were significantly dysregulated in the FF of patients with DOR, demonstrating strong associations with serum AMH and basal estradiol concentrations. A minimized diagnostic panel comprising four core proteins, adenosine deaminase (ADA), vascular endothelial growth factor A (VEGFA), eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1), and matrix metalloproteinase-1 (MMP-1), was established. This multivariable model achieved an excellent area under the receiver operating characteristic curve (AUC) of 0.953. Conclusions: The identified four-protein signature reflects localized chronic inflammation and early pathophysiological shifts in the DOR follicular microenvironment. As a high-performance molecular index, this panel could complement conventional systemic assessments, provide a reliable means of evaluating follicular viability, and optimize individualized therapeutic strategies.<\/jats:p>","DOI":"10.3390\/jcm15114072","type":"journal-article","created":{"date-parts":[[2026,5,25]],"date-time":"2026-05-25T11:57:44Z","timestamp":1779710264000},"page":"4072","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification of Inflammatory Markers for the Prediction and Diagnosis of Diminished Ovarian Reserve Using Olink Targeted Proteomics"],"prefix":"10.3390","volume":"15","author":[{"given":"Meihui","family":"Li","sequence":"first","affiliation":[{"name":"Shanghai-MOST Key Laboratory of Health and Disease Genomics, NHC Key Lab of Reproduction Regulation, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Clinical Medical School, Fudan University, Shanghai 200032, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shanghai-MOST Key Laboratory of Health and Disease Genomics, NHC Key Lab of Reproduction Regulation, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Clinical Medical School, Fudan University, Shanghai 200032, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lin","family":"Yu","sequence":"additional","affiliation":[{"name":"Shanghai-MOST Key Laboratory of Health and Disease Genomics, NHC Key Lab of Reproduction Regulation, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Clinical Medical School, Fudan University, Shanghai 200032, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Shi","sequence":"additional","affiliation":[{"name":"Shanghai-MOST Key Laboratory of Health and Disease Genomics, NHC Key Lab of Reproduction Regulation, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Clinical Medical School, Fudan University, Shanghai 200032, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minzhi","family":"Gao","sequence":"additional","affiliation":[{"name":"Shanghai Key Laboratory for Assisted Reproduction and Reproductive Genetics, Center for Reproductive Medicine, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200135, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nian","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Integrative Medicine, Eastern Hepatobiliary Surgery Hospital, Naval Medical University, Shanghai 200438, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7531-9994","authenticated-orcid":false,"given":"Zhaogui","family":"Sun","sequence":"additional","affiliation":[{"name":"Shanghai-MOST Key Laboratory of Health and Disease Genomics, NHC Key Lab of Reproduction Regulation, Shanghai Institute for Biomedical and Pharmaceutical Technologies, Clinical Medical School, Fudan University, Shanghai 200032, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4282","DOI":"10.1038\/s41467-026-70964-5","article-title":"Modified letrozole vs GnRH antagonist protocols in ovarian aging women for IVF: An open-label, multicenter, randomized controlled trial","volume":"17","author":"Zhao","year":"2026","journal-title":"Nat. Commun."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s13048-024-01584-0","article-title":"The effects of melatonin on follicular oxidative stress and art outcomes in women with diminished ovarian reserve: A randomized controlled trial","volume":"18","author":"Sadeghpour","year":"2025","journal-title":"J. Ovarian Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1186\/s13287-025-04383-6","article-title":"Cell-free fat extract prevents diminished ovarian reserve by inhibiting granulosa cell senescence","volume":"16","author":"Liu","year":"2025","journal-title":"Stem Cell Res. Ther."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Ahmad, M.F., Mohd Nor, N.Y., Mohammad Ramadneh, M.M., Roseli, N.I., Elias, M.H., Mat Jin, N., Abu, M.A., Syafruddin, S.E., Zainuddin, A.A., and Azhar, S.S. (2025). Comparative Analysis of Rescue-In Vitro-Maturation (r-IVM) Outcomes in Women with Diminished Ovarian Reserve (DOR) Versus Normal Ovarian Reserve (NOR). Biomedicines, 13.","DOI":"10.3390\/biomedicines13051084"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"hoae063","DOI":"10.1093\/hropen\/hoae063","article-title":"Women may not benefit from repeated frozen embryo transfers: A retrospective analysis of the cumulative live birth rate of 43972 women","volume":"2024","author":"Zeng","year":"2024","journal-title":"Hum. Reprod. Open"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"(2020). Testing and interpreting measures of ovarian reserve: A committee opinion. Fertil. Steril., 114, 1151\u20131157.","DOI":"10.1016\/j.fertnstert.2020.09.134"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Papini, F., Cappellini, S., Marcacci, I., Marzi, I., Casarosa, E., Daniele, S., Macaluso, S., Ferrari, A., Panattoni, A., and Artini, P.G. (2026). Endometriosis-Related Impairment in Assisted Reproductive Technologies: Inflammatory Profiles, Oocyte Competence, and Embryo Development. J. Clin. Med., 15.","DOI":"10.3390\/jcm15051723"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1186\/s13048-026-02056-3","article-title":"Putative \u201covarian awakening\u201d in diminished ovarian reserve: A multicenter analysis demonstrates indistinguishability from regression to the mean","volume":"19","author":"Huang","year":"2026","journal-title":"J. Ovarian Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1186\/s12958-025-01367-2","article-title":"Potential ability of circulating INSL3 level for the prediction of ovarian reserve and IVF success as a novel theca cell-specific biomarker in women with unexplained infertility and diminished ovarian reserve","volume":"23","author":"Ozcan","year":"2025","journal-title":"Reprod. Biol. Endocrinol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1186\/s13048-023-01293-0","article-title":"Oxidative stress and inflammatory markers in ovarian follicular fluid of women with diminished ovarian reserve during in vitro fertilization","volume":"16","author":"Huang","year":"2023","journal-title":"J. Ovarian Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1186\/s13048-026-01973-7","article-title":"Metabolic dysregulation in follicular fluid is associated with adverse reproductive outcomes in women with decreased ovarian reserve","volume":"19","author":"Li","year":"2026","journal-title":"J. Ovarian Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"109573","DOI":"10.1016\/j.envint.2025.109573","article-title":"Elevated follicular fluid concentrations of ultraviolet filters associated with diminished ovarian reserve: A case-control study","volume":"201","author":"Tian","year":"2025","journal-title":"Environ. Int."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1038\/s42003-026-09826-1","article-title":"Multicellular origins of murine ovarian inflammaging","volume":"9","author":"Galligos","year":"2026","journal-title":"Commun. Biol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"An, X., Guo, X., Cai, M., and Xu, M. (2025). Exploring the Regulatory Effect of Hydroxytyrosol on Ovarian Inflammaging Through Autophagy-Targeted Mechanisms: A Bioinformatics Approach. Nutrients, 17.","DOI":"10.3390\/nu17091421"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e32757","DOI":"10.1097\/MD.0000000000032757","article-title":"The effect of individual oocyte matched follicular fluid oxidant, antioxidant status, and pro- and anti-inflammatory cytokines on IVF outcomes of patients with diminished ovarian reserve","volume":"102","author":"Bacanakgil","year":"2023","journal-title":"Medicine"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1525392","DOI":"10.3389\/fendo.2025.1525392","article-title":"Proximity extension assay revealed novel inflammatory biomarkers for follicular development and ovarian function: A prospective controlled study combining serum and follicular fluid","volume":"16","author":"Wang","year":"2025","journal-title":"Front. Endocrinol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"5597","DOI":"10.2147\/DDDT.S505689","article-title":"Advances in Traditional Chinese Medicine for Managing Diminished Ovarian Reserve: Mechanisms and Clinical Insights","volume":"19","author":"Liu","year":"2025","journal-title":"Drug Des. Devel Ther."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1186\/s12944-024-02295-7","article-title":"Association between the age-adjusted visceral adiposity index (AVAI) and female infertility status: A cross-sectional analysis of the NHANES 2013-2018","volume":"23","author":"Kuang","year":"2024","journal-title":"Lipids Health Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2713","DOI":"10.1038\/s41467-026-69557-z","article-title":"Transdermal testosterone gel vs placebo in women with diminished ovarian reserve prior to in vitro fertilization: A randomized, clinical trial","volume":"17","author":"Polyzos","year":"2026","journal-title":"Nat. Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"110511","DOI":"10.1016\/j.biopha.2020.110511","article-title":"Intermittent hypobaric hypoxia causes deleterious effects on the reproductive system in female rats","volume":"130","author":"Liu","year":"2020","journal-title":"Biomed. Pharmacother."},{"key":"ref_21","first-page":"11635","article-title":"Bisphenol A Alternatives in Follicular Fluid: Novel Risk Factors for Ovarian Reserve Dysfunction","volume":"11","author":"Ren","year":"2026","journal-title":"ACS Omega"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Si, M., Xiong, X., Yun, C., Chen, Y., Niu, H., Qu, Y., Liu, M., Wang, Y., Huang, L., and Long, X. (2026). Microplastics and nanoplastics in follicular fluid are associated with diminished ovarian reserve: Clinical and molecular insights. J. Adv. Res., in press.","DOI":"10.1016\/j.jare.2026.01.074"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Voros, C., Chatzinikolaou, F., Papadimas, G., Polykalas, S., Mavrogianni, D., Koulakmanidis, A.M., Athanasiou, D., Kanaka, V., Kanaka, M., and Bananis, K. (2025). Ferroptosis in the Ovarian Follicular Microenvironment: A Redox-Dependent Cell Death Pathway with Emerging Roles in PCOS, Oocyte Quality, and IVF Outcomes. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms262110381"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Wang, J., Li, Z., Hu, K., Hu, J., Jiang, T., Liao, J., Zhang, Q., Sun, L., Cui, L., and Chen, R. (2026). Exposure to per- and poly-fluoroalkyl substances and associations with embryo quality and adverse pregnancy outcomes: A prospective cohort study. Front. Public Health, 14.","DOI":"10.3389\/fpubh.2026.1783940"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Hall, B., Castelli, L., Higginbottom, A., He, J., Zou, L.N., Walker, H., Yag\u00fce-Capilla, M., Wong, K.E., Burrows, D.J., and George, J. (2026). Antisense Dipeptide Repeat Proteins Drive Widescale Purine Metabolism Aberration in C9orf72 Amyotrophic Lateral Sclerosis via ADA. Int. J. Mol. Sci., 27.","DOI":"10.3390\/ijms27041953"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Ohta, A., and Sitkovsky, M. (2014). Extracellular adenosine-mediated modulation of regulatory T cells. Front. Immunol., 5.","DOI":"10.3389\/fimmu.2014.00304"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"178459","DOI":"10.1016\/j.ejphar.2025.178459","article-title":"GLP-1RA partially alleviates obesity-induced reproductive dysfunction driven by the interplay mechanisms of inflammation and metabolic dysregulation via the SIRT-associated pathway","volume":"1011","author":"Matiki","year":"2026","journal-title":"Eur. J. Pharmacol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1005","DOI":"10.1186\/s12967-025-06966-6","article-title":"Mitochondria-driven inflammation: A new frontier in ovarian ageing","volume":"23","author":"Ju","year":"2025","journal-title":"J. Transl. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1016\/j.mce.2011.04.024","article-title":"Intra-ovarian roles of activins and inhibins","volume":"359","author":"Knight","year":"2012","journal-title":"Mol. Cell. Endocrinol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1677\/jme.1.01921","article-title":"Spatio-temporal analysis of vascular endothelial growth factor expression and blood vessel remodelling in pig ovarian follicles during the periovulatory period","volume":"36","author":"Martelli","year":"2006","journal-title":"J. Mol. Endocrinol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1093\/biolre\/ioae157","article-title":"Single-cell RNA sequencing reveals common interactions between follicle immune cells and granulosa cells in premature ovarian insufficiency patients","volume":"112","author":"Han","year":"2025","journal-title":"Biol. Reprod."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1038\/s41467-024-55509-y","article-title":"Physiological premature aging of ovarian blood vessels leads to decline in fertility in middle-aged mice","volume":"16","author":"Mu","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"110639","DOI":"10.1016\/j.abb.2025.110639","article-title":"Mechanism of vascular endothelial growth factor regulating hypoxia and inflammatory microenvironment in endometriosis: Based on bioinformatics and multi-level validation","volume":"774","author":"Yu","year":"2025","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"669","DOI":"10.1093\/biolreprod\/85.s1.669","article-title":"hVEGFA165b Over-expression Leads to Reduced Ovarian Size and Primordial Follicle Number in Mice","volume":"85","author":"Pohlmeier","year":"2011","journal-title":"Biol. Reprod."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1186\/s13048-023-01228-9","article-title":"Lysophosphatidic acid is associated with oocyte maturation by enhancing autophagy via PI3K-AKT-mTOR signaling pathway in granulosa cells","volume":"16","author":"Liu","year":"2023","journal-title":"J. Ovarian Res."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Li, C., Zhu, W., Ma, X., Fan, X., Ha, F., and Miao, Y. (2026). ESR2 Regulates Granulosa Cell Proliferation and Steroidogenesis via the PI3K\/AKT\/mTOR Signaling Pathway in Wuding Chickens. Biology, 15.","DOI":"10.3390\/biology15040370"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2285","DOI":"10.1128\/MCB.01517-12","article-title":"Inactivation of the mTORC1-eukaryotic translation initiation factor 4E pathway alters stress granule formation","volume":"33","author":"Fournier","year":"2013","journal-title":"Mol. Cell Biol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Wang, X.P., Ye, M.Z., Tu, W.T., and Zhao, X.F. (2025). 20-hydroxyecdysone via upregulating 4EBP expression and inhibiting its phosphorylation represses cell proliferation to permit insect larval molting. Insect Sci., online ahead of print.","DOI":"10.1111\/1744-7917.70025"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"4102","DOI":"10.4049\/jimmunol.1701670","article-title":"eIF4E-Binding Proteins 1 and 2 Limit Macrophage Anti-Inflammatory Responses through Translational Repression of IL-10 and Cyclooxygenase-2","volume":"200","author":"William","year":"2018","journal-title":"J. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"107947","DOI":"10.1016\/j.anireprosci.2025.107947","article-title":"Protective effect of \u03b2-carotene against lipopolysaccharide-induced meiotic maturation defects in porcine cumulus-oocytes complexes","volume":"279","author":"Chen","year":"2025","journal-title":"Anim. Reprod. Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2285","DOI":"10.7150\/ijbs.115990","article-title":"CXCL8 Drives MMP1 Upregulation and Promotes Metastatic Progression in Oral Cancer Through CXCR1\/2-Mediated JAK1\/STAT3 Activation","volume":"22","author":"Lin","year":"2026","journal-title":"Int. J. Biol. Sci."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Verma, S., Lee, I., Schneider, J.S., Farrell, N.F., Kendall, P.L., and Roland, L.T. (2026). The Role of Proteases in Epithelial Dysregulation in Fungal Sinusitis. Int. Forum Allergy Rhinol., online ahead of print.","DOI":"10.1002\/alr.70107"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e70026","DOI":"10.1002\/biof.70026","article-title":"The Potential Role of Matrix Metalloproteinase in Polycystic Ovary Syndrome: Implications for Extracellular Matrix Remodeling","volume":"51","author":"Nikanfar","year":"2025","journal-title":"Biofactors"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Butler, A.E., Nandakumar, M., Sathyapalan, T., Brennan, E., and Atkin, S.L. (2025). Matrix Metalloproteinases, Tissue Inhibitors of Metalloproteinases, and Their Ratios in Women with Polycystic Ovary Syndrome and Healthy Controls. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26010321"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"104450","DOI":"10.1016\/j.rbmo.2024.104450","article-title":"Biomechanical properties of ovarian tissue and their impact on the activation of follicular growth: A narrative review","volume":"50","author":"Vasse","year":"2025","journal-title":"Reprod. Biomed. Online"}],"container-title":["Journal of Clinical Medicine"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2077-0383\/15\/11\/4072\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T04:20:42Z","timestamp":1779855642000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2077-0383\/15\/11\/4072"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,25]]},"references-count":45,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["jcm15114072"],"URL":"https:\/\/doi.org\/10.3390\/jcm15114072","relation":{},"ISSN":["2077-0383"],"issn-type":[{"value":"2077-0383","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,25]]}}}