{"paper_id":"91843cb5-84c4-48dc-8b69-85db0aa0add8","body_text":"Abstract\nEndometriosis is an immune chronic inflammatory disease, and there are currently no more effective drugs for treating endometriosis due to its unknown etiology. Salbutamol is a β2-adrenergic receptor (β2AR) agonist commonly used to treat asthma by selectively activating β2 receptors on airway smooth muscle and leukocytes, exerting bronchial dilation and synergistic anti-inflammatory effects. In recent years, β2AR agonists have been used in endometriosis studies, and we speculate that salbutamol may have a therapeutic effect on endometriosis. The purpose of this research was to explore the therapeutic effect of salbutamol on endometriosis mice. The mouse endometriosis model was established and treated with different doses of salbutamol. Endometrial lesions were harvested for pathological diagnosis, immunohistochemistry (IHC), Masson staining, and toluidine blue analysis. We found that the number and size of endometriotic lesions were all significantly decreased after 3 weeks of treatment with different doses of salbutamol on endometriosis model mice (P < 0.05). After Salbutamol treatment, the amount of mast cells (toluidine blue) and macrophages (F4/80) in the lesions as well as the expressions of interleukin (IL)-1β, tumor necrosis factor (TNF)-ɑ, platelet-derived growth factor subunit B (PDGFB), CD31, transforming growth factor (TGF)-β, Masson staining, BCL2, TUBB3, substance P (SP), and nerve growth factor (NGF) were significantly reduced (P < 0.05). These results suggested that salbutamol could effectively treat endometriosis in mice by reducing immune inflammatory cells and factors, angiogenesis, and fibrosis, increasing apoptosis of endometriotic lesions, and decreasing neurogenesis.\nSimilar content being viewed by others\nData Availability\nThe data in this study are available from the corresponding author upon reasonable request.\nCode Availability\nNot applicable.\nReferences\nTaylor H, Kotlyar A, Flores V. Endometriosis is a chronic systemic disease: clinical challenges and novel innovations. Lancet. 2021;397(10276):839–52.\nBurney R, Giudice L. Pathogenesis and pathophysiology of endometriosis. Fertil Steril. 2012;98(3):511–9.\nKoninckx P, Fernandes R, Ussia A, Schindler L, Wattiez A, Al-Suwaidi S, Amro B, Al-Maamari B, Hakim Z, Tahlak M. Pathogenesis based diagnosis and treatment of endometriosis. Front Endocrinol. 2021;12:745548.\nBurney R. Fibrosis as a molecular hallmark of endometriosis pathophysiology. Fertil Steril. 2022;118(1):203–4.\nBecker C, Bokor A, Heikinheimo O, Horne A, Jansen F, Kiesel L, King K, Kvaskoff M, Nap A, Petersen K, Saridogan E, Tomassetti C, van Hanegem N, Vulliemoz N, Vermeulen N. ESHRE guideline: endometriosis. Hum Reprod Open. 2022;2022(2):hoac009.\nBrichant G, Laraki I, Henry L, Munaut C, Nisolle M. New therapeutics in endometriosis: a review of hormonal, non-hormonal, and non-coding RNA treatments. Int J Mol Sci. 2021;22(19):10498.\nKalaitzopoulos D, Samartzis N, Kolovos G, Mareti E, Samartzis E, Eberhard M, Dinas K, Daniilidis A. Treatment of endometriosis: a review with comparison of 8 guidelines. BMC Womens Health. 2021;21(1):397.\nDeng L, Wang S, Zhang R, Huang J, Lin Y, Liu X, Lu Z, Li M, Tan W. Protective effects of (R)-enantiomers but not (S)-enantiomers of β2-adrenergic receptor agonists against acute colitis: the role of β2AR. Int Immunopharmacol. 2022;110:108997.\nJohnson G, Tabner A, Fakis A, Sherman R, Chester V, Bedford E, Jackson R, Ratan H, Mason S. Salbutamol for analgesia in renal colic: study protocol for a prospective, randomised, placebo-controlled phase II trial (SARC). Trials. 2022;23(1):352.\nFrugier C, Graham F, Samaan K, Paradis L, Des Roches A, Bégin P. Potential efficacy of high-dose inhaled salbutamol for the treatment of abdominal pain during oral food challenge. J Allergy Clin Immunol Pract. 2021;9(8):3130–7.\nSpring Walsh B, Gardiner F, Bloxsome D, Ford D, Mills B, Laws S. A cohort comparison study on women in threatened preterm labor given nifedipine or nifedipine and salbutamol tocolysis in air medical retrieval. Air Méd J. 2022;41(3):298–302.\nDeng L, Guo H, Wang S, Liu X, Lin Y, Zhang R, Tan W. The attenuation of chronic ulcerative colitis by (R)-salbutamol in repeated DSS-induced mice. Oxidative Med Cell Longev. 2022;2022:9318721.\nKim S, Keum B, Byun J, Kim B, Lee K, Yeon J, Lee J, Choi H, Kim E, Jeen Y, Lee H, Chun H, Kim T. Colonic mucosal immune activation in mice with ovalbumin-induced allergic airway disease: association between allergic airway disease and irritable bowel syndrome. Int J Mol Sci. 2021;23(1):181.\nKomlósi Z, van de Veen W, Kovács N, Szűcs G, Sokolowska M, O'Mahony L, Akdis M, Akdis C. Cellular and molecular mechanisms of allergic asthma. Mol Asp Med. 2022;85:100995.\nWasti B, Liu S, Xiang X. Role of epigenetics in the pathogenesis, treatment, prediction, and cellular transformation of asthma. Mediat Inflamm. 2021;2021:9412929.\nMcCallion A, Nasirzadeh Y, Lingegowda H, Miller J, Khalaj K, Ahn S, Monsanto S, Bidarimath M, Sisnett D, Craig A, Young S, Lessey B, Koti M, Tayade C. Estrogen mediates inflammatory role of mast cells in endometriosis pathophysiology. Front Immunol. 2022;13:961599.\nVelho RV, Taube E, Sehouli J, Mechsner S. Neurogenic inflammation in the context of endometriosis-what do we know? Int J Mol Sci. 2021;22(23):13102.\nSprague R, Kim J, Kirimlioglu E, Guo X, Günay N, Guzeloglu-Kayisli O, Ozmen A, Schatz F, Imudia A, Lockwood C, Magness R, Kayisli U. Catecholestradiol activation of adrenergic receptors induces endometrial cell survival via p38 MAPK signaling. J Clin Endocrinol Metab. 2021;106(2):337–50.\nYilmaz N, Ozaksit G, Keskin R, Tapisiz O, Mollamahmutoglu L, Uysal S, Astarci M, Ustun H, Mulazımoglu B. The effect of formoterol on peritoneal VEGF levels in rats with endometriosis. Cytokine. 2012;58(1):47–9.\nZhu J, Liu H, Mao L. The protective effects of ritodrine against hypoxia/reoxygenation-induced injury in endometrial stromal cells. Hum Exp Toxicol. 2022;41:9603271221120650.\nYu Q, Wang J, Li T, Guo X, Ding S, Che X, Zhu L, Peng Y, Xu X, Zou G, Zhang X. Recepteur d’origine nantais contributes to the development of endometriosis via promoting epithelial-mesenchymal transition of a endometrial epithelial cells. J Cell Mol Med. 2021;25(3):1601–12.\nZhu T, Zou G, Ding S, Li T, Zhu L, Wang J, Yao Y, Zhang X. Mast cell stabilizer ketotifen reduces hyperalgesia in a rodent model of surgically induced endometriosis. J Pain Res. 2019;12:1359–69.\nEteraf-Oskouei T, Akbarzadeh-Atashkhosrow A, Maghsudi M, Najafi M. Effects of salbutamol on the inflammatory parameters and angiogenesis in the rat air pouch model of inflammation. Res Pharm Sci. 2017;12(5):364–72.\nRiccio L, Santulli P, Marcellin L, Abrão M, Batteux F, Chapron C. Immunology of endometriosis. Best Pract Res Clin Obstet Gynaecol. 2018;50:39–49.\nTai FW, Chang CY, Chiang JH, Lin WC, Wan L. Association of pelvic inflammatory disease with risk of endometriosis: a nationwide cohort study involving 141,460 individuals. J Clin Med. 2018;7(11):379.\nSadowska A, Manuel-y-Keenoy B, De Backer W. Inhibition of in vitro neutrophil migration through a bilayer of endothelial and epithelial cells using beta2-agonists: concomitant effects on IL-8 and elastase secretion and impact of glucocorticosteroids. Pulm Pharmacol Ther. 2005;18(5):354–62.\nKato T, Yasuda K, Matsushita K, Ishii K, Hirota S, Yoshimoto T, Shibahara H. Interleukin-1/-33 signaling pathways as therapeutic targets for endometriosis. Front Immunol. 2019;10:2021.\nLiu S, Xin X, Hua T, Shi R, Chi S, Jin Z, Wang H. Efficacy of anti-VEGF/VEGFR agents on animal models of endometriosis: a systematic review and meta-analysis. PLoS One. 2016;11(11):e0166658.\nLaschke M, Giebels C, Menger M. Vasculogenesis: a new piece of the endometriosis puzzle. Hum Reprod Update. 2011;17(5):628–36.\nO'Leary A, Fox J, Pullar C. Beta-adrenoceptor activation reduces both dermal microvascular endothelial cell migration via a cAMP-dependent mechanism and wound angiogenesis. J Cell Physiol. 2015;230(2):356–65.\nKitajima M, Dolmans M, Donnez O, Masuzaki H, Soares M, Donnez J. Enhanced follicular recruitment and atresia in cortex derived from ovaries with endometriomas. Fertil Steril. 2014;101(4):1031–7.\nVicino M, Scioscia M, Resta L, Marzullo A, Ceci O, Selvaggi L. Fibrotic tissue in the endometrioma capsule: surgical and physiopathologic considerations from histologic findings. Fertil Steril. 2009;91:1326–8.\nShi L, Zhou F, Zhu H, Huang D, Jin X, Li C, Dai Y, Pan Y, Zhang S. Transforming growth factor beta1 from endometriomas promotes fibrosis in surrounding ovarian tissues via Smad2/3 signaling. Biol Reprod. 2017;97(6):873–82.\nSurinkaew S, Aflaki M, Takawale A, Chen Y, Qi X, Gillis M, Shi Y, Tardif J, Chattipakorn N, Nattel S. Exchange protein activated by cyclic-adenosine monophosphate (Epac) regulates atrial fibroblast function and controls cardiac remodelling. Cardiovasc Res. 2019;115(1):94–106.\nChipuk J, Fisher J, Dillon C, Kriwacki R, Kuwana T, Green D. Mechanism of apoptosis induction by inhibition of the anti-apoptotic BCL-2 proteins. Proc Natl Acad Sci U. S. A. 2008;105(51):20327–32.\nMeresman G, Vighi S, Buquet R, Contreras-Ortiz O, Tesone M, Rumi L. Apoptosis and expression of Bcl-2 and Bax in eutopic endometrium from women with endometriosis. Fertil Steril. 2000;74(4):760–6.\nAjmal I, Farooq M, Abbas S, Shah J, Majid M, Jiang W. Isoprenaline and salbutamol inhibit pyroptosis and promote mitochondrial biogenesis in arthritic chondrocytes by downregulating β-arrestin and GRK2. Front Pharmacol. 2022;13:996321.\nKankaanranta H, Lindsay M, Giembycz M, Zhang X, Moilanen E, Barnes P. Delayed eosinophil apoptosis in asthma. J Allergy Clin Immunol. 2000;106:77–83.\nTse R, Marroquin B, Dorscheid D, White S. Beta-adrenergic agonists inhibit corticosteroid-induced apoptosis of airway epithelial cells. Am J Physiol Lung Cell Mol Physiol. 2003;285(2):L393–404.\nShi Q, Hou Y, Yang Y, Bai G. Protective effects of glycyrrhizin against β2-adrenergic receptor agonist-induced receptor internalization and cell apoptosis. Biol Pharm Bull. 2011;34(5):609–17.\nZaugg M, Xu W, Lucchinetti E, Shafiq S, Jamali N, Siddiqui M. Beta-adrenergic receptor subtypes differentially affect apoptosis in adult rat ventricular myocytes. Circ. 2000;102(3):344–50.\nWang C, Lin H, Lin C, Yu C, Liu S, Huang K, Chung K, Kuo H. Effect of theophylline and specific phosphodiesterase IV inhibition on proliferation and apoptosis of progenitor cells in bronchial asthma. Br J Pharmacol. 2003;138(6):1147–55.\nCoxon L, Horne A, Vincent K. Pathophysiology of endometriosis-associated pain: a review of pelvic and central nervous system mechanisms. Best Pract Res Clin Obstet Gynaecol. 2018;51:53–67.\nYan D, Liu X, Guo S. Neuropeptides substance P and calcitonin gene related peptide accelerate the development and fibrogenesis of endometriosis. Sci Rep. 2019;9(1):2698.\nHonda M, Ito Y, Hattori K, Hosono K, Sekiguchi K, Tsujikawa K, Unno N, Majima M. Inhibition of receptor activity-modifying protein 1 suppresses the development of endometriosis and the formation of blood and lymphatic vessels. J Cell Mol Med. 2020;24(20):11984–97.\nYuan M, Ding S, Meng T, Lu B, Shao S, Zhang X, Yuan H, Hu F. Effect of A-317491 delivered by glycolipid-like polymer micelles on endometriosis pain. Int J Nanomed. 2017;12:8171–83.\nNicholson R, Dixon A, Spanswick D, Lee K. Noradrenergic receptor mRNA expression in adult rat superficial dorsal horn and dorsal root ganglion neurons. Neurosci Lett. 2005;380(3):316–21.\nChoucair-Jaafar N, Yalcin I, Rodeau J, Waltisperger E, Freund-Mercier M, Barrot M. Beta2-adrenoceptor agonists alleviate neuropathic allodynia in mice after chronic treatment. Br J Pharmacol. 2009;158(7):1683–94.\nFunding\nThis work was supported by the National Key R&D Program of China (Grant number: 2017YFC1001202) and the National Natural Science Foundation of China (Grant numbers: 81802591, 81974225, 82171636).\nAuthor information\nAuthors and Affiliations\nContributions\nXZ conceived this experiment. QY and LC contributed equally to conducting experiments, collecting data, and writing the first draft. JW participated in the experimental design and revised the manuscript. XX, TL, XG, MY, and XM participated in the experiment and collected the data. LZ, GZ, and YL conduct statistical analysis on the data.\nCorresponding author\nEthics declarations\nEthics Approval\nOur animal experiment program has been reviewed and approved by the Ethics Committee of Zhejiang University (ZJU20220121).\nConsent to Participate\nNot applicable.\nConsent for publication\nNot applicable.\nConflict of Interest\nThe authors declare no competing interests.\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\nYu, Q., Chen, L., Wang, J. et al. Investigation of the Therapeutic Effect of Salbutamol on Endometriosis in a Mouse Model. Reprod. Sci. 31, 430–440 (2024). https://doi.org/10.1007/s43032-023-01371-0\nReceived:\nAccepted:\nPublished:\nVersion of record:\nIssue date:\nDOI: https://doi.org/10.1007/s43032-023-01371-0","source_license":"CC0","license_restricted":false}