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Jellyfish-derived peptides have recently emerged as promising marine bioactive compounds with potential anti-inflammatory activity. In this study, three candidate anti-inflammatory peptides were identified from the enzymatic hydrolysate of Nemopilema nomurai through an integrated strategy combining LC\u2013MS\/MS-based peptidomics and multi-step in silico screening. The selected peptides (DGIPGMPG, PGFHVPPP, and GPKGYPGP) were prioritized based on predicted bioactivity, non-toxicity, favorable physicochemical properties, and molecular docking with the TLR4\/MD-2\/LPS complex (PDB ID: 3FXI), with docking scores ranging from \u22128.4 to \u22128.3 kcal\/mol. Subsequent experimental validation demonstrated that all three peptides exhibited good cytocompatibility and significantly inhibited LPS-induced nitric oxide (NO) production in RAW264.7 macrophages, with GPKGYPGP showing the strongest effect. In addition, these peptides effectively reduced the secretion of pro-inflammatory cytokines, including TNF-\u03b1 and IL-1\u03b2, to varying extents. Collectively, this study identifies three novel anti-inflammatory peptides derived from jellyfish enzymatic hydrolysates and highlights their potential as promising candidates for the development of marine-derived bioactive agents targeting inflammation-related diseases.<\/jats:p>","DOI":"10.3390\/md24060192","type":"journal-article","created":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T09:57:43Z","timestamp":1779962263000},"page":"192","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification of Novel Anti-Inflammatory Peptides from Jellyfish Nemopilema nomurai Enzymatic Hydrolysate: An Integrated In Silico Analysis and Cellular Evaluation"],"prefix":"10.3390","volume":"24","author":[{"given":"Wen","family":"Shen","sequence":"first","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266000, China"},{"name":"University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2266-2908","authenticated-orcid":false,"given":"Xueqin","family":"Wang","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266000, China"},{"name":"Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao 266237, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rongfeng","family":"Li","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266000, China"},{"name":"Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao 266237, China"},{"name":"University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Song","family":"Liu","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266000, China"},{"name":"Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao 266237, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronge","family":"Xing","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266000, China"},{"name":"Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao 266237, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengcheng","family":"Li","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3213-6212","authenticated-orcid":false,"given":"Huahua","family":"Yu","sequence":"additional","affiliation":[{"name":"Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266000, China"},{"name":"Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao 266237, China"},{"name":"University of Chinese Academy of Sciences, Beijing 100049, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"730","DOI":"10.1016\/j.jconrel.2024.06.063","article-title":"Recent Advances in Targeted Therapy for Inflammatory Vascular Diseases","volume":"372","author":"Zhao","year":"2024","journal-title":"J. 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