{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T15:41:19Z","timestamp":1785426079622,"version":"3.56.0"},"reference-count":28,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T00:00:00Z","timestamp":1779926400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32172899"],"award-info":[{"award-number":["32172899"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"award":["32172899"],"award-info":[{"award-number":["32172899"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Chronic kidney disease (CKD) represents a considerable health burden for both humans and veterinary patients globally. Renal fibrosis is the final common pathway for the progression of CKD to end-stage renal disease, which can eventually lead to renal failure. Thymoquinone (TQ), the primary bioactive constituent of Nigella sativa, has demonstrated significant antifibrotic potential; however, the specific molecular mechanisms underlying its renoprotective effects remain incompletely elucidated. This study aimed to investigate how TQ alleviates renal fibrosis to support its potential as a therapeutic agent. TQ\u2019s renoprotective effects were evaluated in a murine unilateral ureteral obstruction (UUO) model using histopathology, Western blotting, immunofluorescence, and RT-qPCR. Network pharmacology and untargeted metabolomics were integrated to identify key pathways, which were further assessed through in vivo and in vitro experiments. TQ treatment attenuated UUO-induced renal interstitial injury. TQ treatment downregulated \u03b1-smooth muscle actin (\u03b1-SMA) and fibronectin, thereby suppressing myofibroblast activation and extracellular matrix (ECM) accumulation. Integrated multi-omics analyses indicated that the antifibrotic activity of TQ is associated with modulation of the PI3K\/AKT signaling axis. Subsequent in vivo and in vitro studies suggested that TQ protects against renal injury by inhibiting aberrant PI3K\/AKT signaling. This study found that TQ ameliorates renal interstitial fibrosis in UUO mice. The underlying mechanism appears to involve suppression of myofibroblast activation and ECM accumulation via inhibition of PI3K\/AKT signaling. These findings highlight the therapeutic potential of TQ for renal fibrosis.<\/jats:p>","DOI":"10.3390\/ijms27114901","type":"journal-article","created":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T12:45:08Z","timestamp":1780404308000},"page":"4901","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Integrating Metabolomics and Network Pharmacology to Reveal the Mechanism of Thymoquinone Alleviating Renal Interstitial Fibrosis in UUO Mice"],"prefix":"10.3390","volume":"27","author":[{"given":"Yuanqing","family":"Liu","sequence":"first","affiliation":[{"name":"College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huijing","family":"Zhang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanjun","family":"Dong","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gebin","family":"Li","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1934-7111","authenticated-orcid":false,"given":"Kai","family":"Fan","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9516-6270","authenticated-orcid":false,"given":"Zhihui","family":"Hao","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"},{"name":"Innovation Centre of Chinese Veterinary Medicine, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"},{"name":"Key Biology Laboratory of Chinese Veterinary Medicine, Ministry of Agriculture and Rural Affairs, Beijing 100193, China"},{"name":"State Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuaiyu","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"},{"name":"Innovation Centre of Chinese Veterinary Medicine, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China"},{"name":"Key Biology Laboratory of Chinese Veterinary Medicine, Ministry of Agriculture and Rural Affairs, Beijing 100193, China"},{"name":"State Key Laboratory of Veterinary Public Health and Safety, College of Veterinary Medicine, China Agricultural University, Beijing 100193, 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":"814","DOI":"10.1111\/jvim.12090","article-title":"Chronic kidney disease in dogs in UK veterinary practices: Prevalence, risk factors, and survival","volume":"27","author":"Elliott","year":"2013","journal-title":"J. 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