{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,31]],"date-time":"2026-03-31T05:32:53Z","timestamp":1774935173567,"version":"3.50.1"},"reference-count":41,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2025,9,13]],"date-time":"2025-09-13T00:00:00Z","timestamp":1757721600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"Ministry of Education","doi-asserted-by":"publisher","award":["RS-2018-NR031048"],"award-info":[{"award-number":["RS-2018-NR031048"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Atopic dermatitis is a chronic inflammatory skin disorder characterized by persistent inflammation and severe pruritus. Current anti-inflammatory agents carry risks of long-term adverse effects, while antihistamines provide limited relief of pruritus. Protease-activated receptor 2 (PAR2) has emerged as a critical mediator of both inflammation and pruritus, representing a promising therapeutic target. In this study, we investigated the therapeutic potential of punicalagin (PCG), a potent PAR2 antagonist, in atopic dermatitis. PCG fully and potently inhibited trypsin-induced PAR2 activation in HaCaT cells with an IC50 of 1.30 \u00b5M, exhibiting over 40-fold greater selectivity over PAR1. PCG significantly inhibited PAR2-induced phosphorylation of ERK1\/2 and NF-\u03baB in both HaCaT and human dermal fibroblast cells and reduced IL-8 secretion in HaCaT cells. In addition, PCG did not significantly affect other pruritus-related GPCRs including H1R, H4R, TGR5, 5HT2A, 5HT2B, and MRGPRX2 at 30 \u00b5M. Notably, PCG strongly blocked PAR2-AP-induced scratching in mice. In addition, PCG improved skin lesions, reduced dermatitis severity scores, and alleviated scratching behavior in a DNFB-induced atopic dermatitis model. These effects were associated with reduced epidermal thickness, decreased serum TSLP levels, and inhibition of PAR2-dependent calcium signaling in dorsal root ganglion neurons. These findings demonstrate that PCG is a selective PAR2 antagonist that effectively alleviates both inflammatory and pruritic symptoms of atopic dermatitis, suggesting its potential as a novel therapeutic agent.<\/jats:p>","DOI":"10.3390\/ijms26188920","type":"journal-article","created":{"date-parts":[[2025,9,16]],"date-time":"2025-09-16T07:33:02Z","timestamp":1758007982000},"page":"8920","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Blockade of PAR2 Signaling by Punicalagin as a Therapeutic Strategy for Atopic Dermatitis"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0711-0766","authenticated-orcid":false,"given":"Hyejin","family":"Jeon","sequence":"first","affiliation":[{"name":"College of Pharmacy and Yonsei Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9335-8604","authenticated-orcid":false,"given":"Yohan","family":"Seo","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Yonsei Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"},{"name":"Department of Bio-Nanomaterials, Bio Campus of Korea Polytechnics, 112 Dongan-ro, Ganggeong-up, Nonsan 32943, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3676-0170","authenticated-orcid":false,"given":"Wook-Joo","family":"Lee","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Gachon University, 191 Hambangmoero, Yeonsu-gu, Incheon 21936, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3780-7695","authenticated-orcid":false,"given":"Yunkyung","family":"Heo","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Yonsei Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Won-Sik","family":"Shim","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Gachon University, 191 Hambangmoero, Yeonsu-gu, Incheon 21936, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1480-7433","authenticated-orcid":false,"given":"Wan","family":"Namkung","sequence":"additional","affiliation":[{"name":"College of Pharmacy and Yonsei Institute of Pharmaceutical Sciences, Yonsei University, 85 Songdogwahak-ro, Yeonsu-gu, Incheon 21983, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"523","DOI":"10.1046\/j.1365-2133.2001.04078.x","article-title":"Prevalence of atopic dermatitis, asthma, allergic rhinitis, and hand and contact dermatitis in adolescents. The Odense Adolescence Cohort Study on Atopic Diseases and Dermatitis","volume":"144","author":"Mortz","year":"2001","journal-title":"Br. J. Dermatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1001\/archdermatol.2011.178","article-title":"The Impact of Pruritus on Quality of Life: The Skin Equivalent of Pain","volume":"147","author":"Kini","year":"2011","journal-title":"Arch. Dermatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1449","DOI":"10.1038\/ng.3424","article-title":"Multi-ancestry genome-wide association study of 21,000 cases and 95,000 controls identifies new risk loci for atopic dermatitis","volume":"47","author":"Paternoster","year":"2015","journal-title":"Nat. Genet."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"786","DOI":"10.1111\/j.0022-202X.2005.23651.x","article-title":"Interleukin-4 Suppresses the Enhancement of Ceramide Synthesis and Cutaneous Permeability Barrier Functions Induced by Tumor Necrosis Factor-\u03b1 and interferon-\u03b3 in Human Epidermis","volume":"124","author":"Hatano","year":"2005","journal-title":"J. Investig. Dermatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1038\/jid.2012.239","article-title":"TSLP Produced by Keratinocytes Promotes Allergen Sensitization through Skin and Thereby Triggers Atopic March in Mice","volume":"133","author":"Hener","year":"2013","journal-title":"J. Investig. Dermatol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"980","DOI":"10.1016\/j.it.2018.10.001","article-title":"The Itch\u2013Scratch Cycle: A Neuroimmune Perspective","volume":"39","author":"Mack","year":"2018","journal-title":"Trends Immunol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1016\/j.jaad.2016.02.1228","article-title":"Systematic review and meta-analysis of randomized clinical trials (RCTs) comparing topical calcineurin inhibitors with topical corticosteroids for atopic dermatitis: A 15-year experience","volume":"75","author":"Broeders","year":"2016","journal-title":"J. Am. Acad. Dermatol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1038\/s41573-021-00266-6","article-title":"Atopic dermatitis: An expanding therapeutic pipeline for a complex disease","volume":"21","author":"Bieber","year":"2021","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1053","DOI":"10.1016\/j.jaip.2020.11.034","article-title":"Biologics for Treatment of Atopic Dermatitis: Current Status and Future Prospect","volume":"9","author":"Ratchataswan","year":"2021","journal-title":"J. Allergy Clin. Immunol. Pr."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1111\/j.1365-2133.1990.tb14732.x","article-title":"The antipruritic effect of a sedative and a non-sedative antihistamine in atopic dermatitis","volume":"122","author":"Wahlgren","year":"1990","journal-title":"Br. J. Dermatol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1930","DOI":"10.1038\/jid.2008.13","article-title":"Mite and Cockroach Allergens Activate Protease-Activated Receptor 2 and Delay Epidermal Permeability Barrier Recovery","volume":"128","author":"Jeong","year":"2008","journal-title":"J. Investig. Dermatol."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Fan, M., Fan, X., Lai, Y., Chen, J., Peng, Y., Peng, Y., Xiang, L., and Ma, Y. (2024). Protease-Activated Receptor 2 in inflammatory skin disease: Current evidence and future perspectives. Front. Immunol., 15.","DOI":"10.3389\/fimmu.2024.1448952"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1016\/0092-8674(91)90261-V","article-title":"Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation","volume":"64","author":"Vu","year":"1991","journal-title":"Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"32095","DOI":"10.1074\/jbc.M513138200","article-title":"Proteinase-activated Receptors, Targets for Kallikrein Signaling","volume":"281","author":"Oikonomopoulou","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Buhl, T., Ikoma, A., Kempkes, C., Cevikbas, F., Sulk, M., Buddenkotte, J., Akiyama, T., Crumrine, D., Camerer, E., and Carstens, E. (2020). Protease-Activated Receptor-2 Regulates Neuro-Epidermal Communication in Atopic Dermatitis. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.01740"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"6176","DOI":"10.1523\/JNEUROSCI.23-15-06176.2003","article-title":"Proteinase-Activated Receptor-2 Mediates Itch: A Novel Pathway for Pruritus in Human Skin","volume":"23","author":"Steinhoff","year":"2003","journal-title":"J. Neurosci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"808","DOI":"10.3349\/ymj.2010.51.6.808","article-title":"Protease and protease-activated receptor-2 signaling in the pathogenesis of atopic dermatitis","volume":"51","author":"Lee","year":"2010","journal-title":"Yonsei Med. J."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1254\/jphs.08332FP","article-title":"Involvement of Tryptase and Proteinase-Activated Receptor-2 in Spontaneous Itch-Associated Response in Mice With Atopy-like Dermatitis","volume":"109","author":"Tsujii","year":"2009","journal-title":"J. Pharmacol. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Nomura, H., Suganuma, M., Takeichi, T., Kono, M., Isokane, Y., Sunagawa, K., Kobashi, M., Sugihara, S., Kajita, A., and Miyake, T. (2020). Multifaceted Analyses of Epidermal Serine Protease Activity in Patients with Atopic Dermatitis. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21030913"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1016\/j.ejphar.2005.11.012","article-title":"Scratching behavior in mice induced by the proteinase-activated receptor-2 agonist, SLIGRL-NH2","volume":"530","author":"Shimada","year":"2006","journal-title":"Eur. J. Pharmacol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2176","DOI":"10.1152\/jn.00463.2009","article-title":"Excitation of Mouse Superficial Dorsal Horn Neurons by Histamine and\/or PAR-2 Agonist: Potential Role in Itch","volume":"102","author":"Akiyama","year":"2009","journal-title":"J. Neurophysiol."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Seo, Y., Mun, C.H., Park, S.-H., Jeon, D., Kim, S.J., Yoon, T., Ko, E., Jo, S., Park, Y.-B., and Namkung, W. (2020). Punicalagin Ameliorates Lupus Nephritis via Inhibition of PAR2. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21144975"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1124\/pr.54.2.203","article-title":"International union of pharmacology. XXVIII. Proteinase-activated receptors","volume":"54","author":"Hollenberg","year":"2002","journal-title":"Pharmacol. Rev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1221","DOI":"10.1038\/sj.gt.3301724","article-title":"Prevention and regression of atopic dermatitis by ointment containing NF-kB decoy oligodeoxynucleotides in NC\/Nga atopic mouse model","volume":"9","author":"Nakamura","year":"2002","journal-title":"Gene Ther."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Zeze, N., Kido-Nakahara, M., Tsuji, G., Maehara, E., Sato, Y., Sakai, S., Fujishima, K., Hashimoto-Hachiya, A., Furue, M., and Nakahara, T. (2022). Role of ERK Pathway in the Pathogenesis of Atopic Dermatitis and Its Potential as a Therapeutic Target. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23073467"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1136\/adc.70.2.119","article-title":"Detection of plasma interleukin-8 in atopic dermatitis","volume":"70","author":"Kimata","year":"1994","journal-title":"Arch. Dis. Child."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"832","DOI":"10.1111\/j.0022-202X.2004.23445.x","article-title":"Protease-Activated Receptor-2 (PAR-2) Expression in Human Fibroblasts is Regulated by Growth Factors and Extracellular Matrix","volume":"123","author":"Gruber","year":"2004","journal-title":"J. Investig. Dermatol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Liu, Q., Li, M., Wang, N., He, C., Jiang, X., and Li, J. (2022). Calcium-Based Antimicrobial Peptide Compounds Attenuate DNFB-Induced Atopic Dermatitis-Like Skin Lesions via Th-Cells in BALB\/c Mice. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms231911371"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"14795","DOI":"10.1073\/pnas.0507385102","article-title":"Retinoid X receptor ablation in adult mouse keratinocytes generates an atopic dermatitis triggered by thymic stromal lymphopoietin","volume":"102","author":"Li","year":"2005","journal-title":"Proc. Natl. Acad. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/j.cell.2013.08.057","article-title":"The Epithelial Cell-Derived Atopic Dermatitis Cytokine TSLP Activates Neurons to Induce Itch","volume":"155","author":"Wilson","year":"2013","journal-title":"Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1016\/j.jid.2018.08.019","article-title":"PAR2 Pepducin-Based Suppression of Inflammation and Itch in Atopic Dermatitis Models","volume":"139","author":"Barr","year":"2019","journal-title":"J. Investig. Dermatol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2525","DOI":"10.1042\/BST20200191","article-title":"The development of proteinase-activated receptor-2 modulators and the challenges involved","volume":"48","author":"McIntosh","year":"2020","journal-title":"Biochem. Soc. Trans."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1067\/mai.2001.118796","article-title":"Targeting keratinocyte apoptosis in the treatment of atopic dermatitis and allergic contact dermatitis","volume":"108","author":"Trautmann","year":"2001","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1413","DOI":"10.1111\/j.1476-5381.2011.01610.x","article-title":"Modulating human proteinase activated receptor 2 with a novel antagonist (GB88) and agonist (GB110)","volume":"165","author":"Suen","year":"2011","journal-title":"Br. J. Pharmacol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1111\/j.1600-0625.1999.tb00383.x","article-title":"Proteinase-activated receptor-2 in human skin: Tissue distribution and activation of keratinocytes by mast cell tryptase","volume":"8","author":"Steinhoff","year":"1999","journal-title":"Exp. Dermatol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1979","DOI":"10.1172\/JCI30951","article-title":"Sensitization of TRPA1 by PAR2 contributes to the sensation of inflammatory pain","volume":"117","author":"Dai","year":"2007","journal-title":"J. Clin. Investig."},{"key":"ref_37","first-page":"e66","article-title":"A Re-Innervated in Vitro Skin Model of Non-Histaminergic Itch and Skin Neurogenic Inflammation: PAR2-, TRPV1- and TRPA1-Agonist Induced Functionality","volume":"1","author":"Lebonvallet","year":"2021","journal-title":"Ski. Heal. Dis."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Xu, J., Cao, K., Liu, X., Zhao, L., Feng, Z., and Liu, J. (2021). Punicalagin Regulates Signaling Pathways in Inflammation-Associated Chronic Diseases. Antioxidants, 11.","DOI":"10.3390\/antiox11010029"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s42003-020-01504-0","article-title":"Protease-activated receptor-2 ligands reveal orthosteric and allosteric mechanisms of receptor inhibition","volume":"3","author":"Kennedy","year":"2020","journal-title":"Commun. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1186\/1476-9255-5-9","article-title":"Bioavailable constituents\/metabolites of pomegranate (Punica granatum L) preferentially inhibit COX2 activity ex vivo and IL-1beta-induced PGE2 production in human chondrocytes i n vitro","volume":"5","author":"Shukla","year":"2008","journal-title":"J. Inflamm."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Shabir, I., Dar, A.H., Dash, K.K., Manzoor, S., Srivastava, S., Pandey, V.K., Shams, R., Bashir, I., Khan, S.A., and Mukarram, S.A. (2024). Bioactive potential of punicalagin: A comprehensive review. Appl. Food Res., 4.","DOI":"10.1016\/j.afres.2024.100572"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/18\/8920\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:45:01Z","timestamp":1760035501000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/18\/8920"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,13]]},"references-count":41,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["ijms26188920"],"URL":"https:\/\/doi.org\/10.3390\/ijms26188920","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,13]]}}}