{"paper_id":"f95b0247-9f6d-4af1-96a5-32e9156a25bc","body_text":"Abstract\nCarvacrol is a phenolic monoterpenoid found in essential oils of oregano, thyme, and other plants. It possesses pharmacological activities, including antioxidant, anticancer, and anti-inflammatory properties, against several diseases. However, research on the treatment effect of endometriosis has not yet been conducted. Endometriosis is a condition characterized by tissues resembling the endometrium beyond the confines of the uterus, leading to intense pelvic pain and heightened risks of infertility. This study aimed to confirm the effectiveness of carvacrol as a treatment strategy for endometriosis with inflammatory regulation and antioxidant property. In the in vivo study, C57BL/6 J mice were orally administered 100 mg/kg carvacrol for 4 weeks and confirmed a decreased size of endometriosis lesions in the carvacrol-treated group, increased number of T cells in the spleen, and increased expression of cytokine-related genes in both tissues. Immortalized human ovarian endometriotic stromal cells (ihOESCs), derived from endometriotic lesions in patients, were used as an in vitro model for studying endometriosis pathophysiology. The findings of the in vitro study confirmed that carvacrol regulates calcium homeostasis, induces antioxidant effects, increases inflammatory response, deactivates the phosphatidylinositol 3-kinase (PI3K) signaling pathway, and ultimately induces autophagy. Therefore, we determined that carvacrol is suitable as an anti-inflammatory and anti-oxidant treatment strategy for endometriosis.\nGraphical Abstract\nSimilar content being viewed by others\nData Availability\nData available on request from the authors.\nCode Availability\nNot applicable.\nReferences\nMehedintu C, Plotogea MN, Ionescu S, Antonovici M. Endometriosis still a challenge. J Med Life. 2014;7:349–57.\nSorrentino F, M DEP, Falagario M, D'Alteri OM, A DISS, Pacheco LA, Carugno JT, Nappi L. Endometriosis and adverse pregnancy outcome. Minerva Obstet Gynecol 2022;74:31–44.\nWang H, Wang B, Wu M, Lu J, Duan P. Targeting osteopontin alleviates endometriosis and inflammation by inhibiting the RhoA/ROS axis and achieves non-invasive in vitro detection via menstrual blood. Hum Reprod. 2024;39:1057–71.\nZhou WJ, Yang HL, Shao J, Mei J, Chang KK, Zhu R, Li MQ. Anti-inflammatory cytokines in endometriosis. Cell Mol Life Sci. 2019;76:2111–32.\nAssaf L, Eid AA, Nassif J. Role of AMPK/mTOR, mitochondria, and ROS in the pathogenesis of endometriosis. Life Sci. 2022;306:120805.\nSingh J, Luqman S, Meena A. Carvacrol as a prospective regulator of cancer targets/signalling pathways. Curr Mol Pharmacol. 2023;16:542–58.\nElbe H, Yigitturk G, Cavusoglu T, Baygar T, OzgulOnal M, Ozturk F. Comparison of ultrastructural changes and the anticarcinogenic effects of thymol and carvacrol on ovarian cancer cells: which is more effective? Ultrastruct Pathol. 2020;44:193–202.\nKianmehr M, Rezaee A, Mahmoudi M, Ghorani V, Boskabady MH. T helper cells subtypes and their cytokine gene expression affected by carvacrol in sensitized mice administered during sensitization period. J Cell Biochem. 2019;120:5343–54.\nKhazdair MR, Ghorani V, Boskabady MH. Experimental and clinical evidence on the effect of carvacrol on respiratory, allergic, and immunologic disorders: A comprehensive review. BioFactors. 2022;48:779–94.\nSon D, Park H, An G, Park S, Hwang DW, Park SJ, Kim HS, Lim W, You S, Song G. Establishment of immortalized human endometriotic stromal cell line from ectopic lesion of a patient with endometriosis. Reprod Sci. 2023;30:2703–14.\nPark W, Park S, Lim W, Song G. Bifenthrin reduces pregnancy potential via induction of oxidative stress in porcine trophectoderm and uterine luminal epithelial cells. Sci Total Environ. 2021;784:147143.\nPelch KE, Sharpe-Timms KL, Nagel SC. Mouse model of surgically-induced endometriosis by auto-transplantation of uterine tissue. Jove-J Vis Exp. 2012;6:(59):e3396.\nPark S, Ham J, Yang CW, Park W, Park H, An GR, Song JS, Hong TY, Park SJ, Kim HS, Song GH, Lim W. Melatonin inhibits endometriosis development by disrupting mitochondrial function and regulating tiRNAs. J Pineal Res 2023;74.\nPark W, Song G, Lim W, Park S. Therapeutic effects of S-allyl-L-cysteine in a mouse endometriosis model and its immunomodulatory effects via regulation of T cell subsets and cytokine expression. Pharmacol Rep. 2024;76:1089–99.\nHerreros-Villanueva M, Chen CC, Tsai EM, Er TK. Endometriosis-associated ovarian cancer: What have we learned so far? Clin Chim Acta. 2019;493:63–72.\nNasiri-Ansari N, Nikolopoulou C, Papoutsi K, Kyrou I, Mantzoros CS, Kyriakopoulos G, Chatzigeorgiou A, Kalotychou V, Randeva MS, Chatha K, Kontzoglou K, Kaltsas G, et al. Empagliflozin attenuates non-alcoholic fatty liver disease (NAFLD) in high fat diet fed ApoE((-/-)) mice by activating autophagy and reducing ER stress and apoptosis. Int J Mol Sci. 2021;22.\nMoloney JN, Cotter TG. ROS signalling in the biology of cancer. Semin Cell Dev Biol. 2018;80:50–64.\nMatsuda J, Namba T, Takabatake Y, Kimura T, Takahashi A, Yamamoto T, Minami S, Sakai S, Fujimura R, Kaimori JY, Matsui I, Hamano T, et al. Antioxidant role of autophagy in maintaining the integrity of glomerular capillaries. Autophagy. 2018;14:53–65.\nXu YJ, Jiang CY, Cheng ZQ, Yao WG, Ge SC. Combining use of phillyrin and autophagy blocker exerts suppressive effect on nasopharyngeal carcinoma cell malignancy and autophagy via AMPK/mTOR/p70s6k signaling pathway. Mol Cell Toxicol. 2024;20:611–8.\nZhao J, He SL, Xiang CH, Zhang SL, Chen XY, Lu XY, Yao Q, Yang LP, Ma LM, Tian WW. KLF9 promotes autophagy and apoptosis in T-cell acute lymphoblastic leukemia cells by inhibiting AKT/mTOR signaling pathway. Mol Cell Toxicol. 2023;19:531–8.\nYu JS, Cui W. Proliferation, survival and metabolism: the role of PI3K/AKT/mTOR signalling in pluripotency and cell fate determination. Development. 2016;143:3050–60.\nChen SL, Yu WY, Li ZY, Wang YD, Peng B. STYXL1 promotes proliferation and epithelial mesenchymal transition of gastric cancer cells via activating the PI3K/AKT pathway. Mol Cell Toxicol. 2024;20:281–91.\nYin H, Zuo Z, Yang Z, Guo H, Fang J, Cui H, Ouyang P, Chen X, Chen J, Geng Y, Chen Z, Huang C, et al. Nickel induces autophagy via PI3K/AKT/mTOR and AMPK pathways in mouse kidney. Ecotoxicol Environ Saf. 2021;223:112583.\nSikora JP, Karawani J, Sobczak J. Neutrophils and the systemic inflammatory response syndrome (SIRS). Int J Mol Sci 2023; 24.\nWynn TA. IL-13 effector functions. Annu Rev Immunol. 2003;21:425–56.\nSilva TA, Ribeiro FL, Oliveira-Neto HH, Watanabe S, AlencarRde C, Fukada SY, Cunha FQ, Leles CR, Mendonca EF, Batista AC. Dual role of CCL3/CCR1 in oral squamous cell carcinoma: implications in tumor metastasis and local host defense. Oncol Rep. 2007;18:1107–13.\nQuast I, Dvorscek AR, Pattaroni C, Steiner TM, McKenzie CI, Pitt C, O’Donnell K, Ding Z, Hill DL, Brink R, Robinson MJ, Zotos D, et al. Interleukin-21, acting beyond the immunological synapse, independently controls T follicular helper and germinal center B cells. Immunity. 2022;55(1414–1430):e1415.\nRebe C, Ghiringhelli F. Interleukin-1beta and cancer. Cancers (Basel) 2020;12.\nMebius RE, Kraal G. Structure and function of the spleen. Nat Rev Immunol. 2005;5:606–16.\nLi N. CD4+ T cells in atherosclerosis: regulation by platelets. Thromb Haemost. 2013;109:980–90.\nWang W, Green M, Choi JE, Gijon M, Kennedy PD, Johnson JK, Liao P, Lang X, Kryczek I, Sell A, Xia H, Zhou J, et al. CD8(+) T cells regulate tumour ferroptosis during cancer immunotherapy. Nature. 2019;569:270–4.\nSalah FS, Ebbinghaus M, Muley VY, Zhou Z, Al-Saadi KR, Pacyna-Gengelbach M, O’Sullivan GA, Betz H, Konig R, Wang ZQ, Brauer R, Petersen I. Tumor suppression in mice lacking GABARAP, an Atg8/LC3 family member implicated in autophagy, is associated with alterations in cytokine secretion and cell death. Cell Death Dis. 2016;7:e2205.\nAcknowledgements\nThis research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (grant number: HI22 C1424). And this research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (grant number: RS- 2023 - 00246500)\nAuthor information\nAuthors and Affiliations\nContributions\nHyewon Jang: Investigation, Methodology, Software, Writing-Original draft preparation; Wonhyoung Park: Investigation, Methodology, Software, Writing-Original draft preparation; Hee Seung Kim: Methodology, Validation; Gwonhwa Song: Conceptualization, Validation, Supervision; Whasun Lim: Validation, Supervision, Writing- Review & Editing; Sunwoo Park: Conceptualization, Writing- Review & Editing, Funding Acquisition.\nCorresponding authors\nEthics declarations\nEthics Approval\nThe animal experiments were conducted according to the approval of the IACUC at Gyeongsang National University (GNU- 220314-M0027).\nConsent to Participate\nNot applicable.\nConsent for Publication\nNot applicable.\nConflict of Interest\nWe confirm that there are no known conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome. We confirm that the manuscript has been read and approved by all named authors and that there are no other persons who satisfied the criteria for authorship but are not listed.\nAdditional information\nPublisher's Note\nSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.\nRights and permissions\nSpringer 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.\nAbout this article\nCite this article\nJang, H., Park, W., Kim, H.S. et al. Carvacrol Shows Potential as a Treatment for Endometriosis with Inflammatory Reaction and Antioxidant Properties, Demonstrated in both In Vivo and In Vitro Studies. Reprod. Sci. 32, 1825–1839 (2025). https://doi.org/10.1007/s43032-025-01866-y\nReceived:\nAccepted:\nPublished:\nVersion of record:\nIssue date:\nDOI: https://doi.org/10.1007/s43032-025-01866-y","source_license":"CC0","license_restricted":false}