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However, its untreatable hepatotoxic effect severely limits its clinical use. Therefore, further studies are required to combat MTX-induced liver injury. Nitrate, abundant in green vegetables, possesses anti-inflammatory, antioxidant, and immunoregulatory effects. Our study investigated the preventive effects of nitrate on MTX-induced liver injury. Liver injury in mice was induced by administering a single dose of MTX (20\u00a0mg\/kg body weight) intraperitoneally (i.p.). Pre-treatment of mice with 2\u00a0mM nitrate in drinking water 5\u00a0days prior effectively mitigated the MTX-elevated serum aminotransferase activities, attenuated hepatic pathological injury, reduced hepatic apoptosis and restored the proliferative capacity of hepatocytes. RNA sequencing analysis indicated that the molecular mechanism may involve the activation of the Wnt\/\u03b2-catenin pathway. The AML 12 cell line was employed for in vitro validation. In this study, the hepatoprotective effect of nitrate against drug-induced liver injury (DILI) was identified for the first time, providing a new approach to preventing DILI in clinical practice.<\/jats:p>","DOI":"10.1093\/toxres\/tfaf107","type":"journal-article","created":{"date-parts":[[2025,7,17]],"date-time":"2025-07-17T10:04:37Z","timestamp":1752746677000},"source":"Crossref","is-referenced-by-count":1,"title":["Nitrate protects against methotrexate-induced liver injury by activating Wnt\/\u03b2-catenin Signaling in mice"],"prefix":"10.1093","volume":"14","author":[{"given":"Xin","family":"Wen","sequence":"first","affiliation":[{"name":"Salivary Gland Disease Center and Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Beijing Laboratory of Oral Health and Beijing Stomatological Hospital, Capital Medical University , No. 9 Fanjiacun Rd, Fengtai District, Beijing 100070 ,","place":["China"]},{"name":"Immunology Research Center for Oral and Systemic Health, Beijing Friendship Hospital, Capital Medical University , No. 95 Yong'an Rd, Xicheng District, Beijing 100050 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ying","family":"Liu","sequence":"additional","affiliation":[{"name":"Salivary Gland Disease Center and Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Beijing Laboratory of Oral Health and Beijing Stomatological Hospital, Capital Medical University , No. 9 Fanjiacun Rd, Fengtai District, Beijing 100070 ,","place":["China"]},{"name":"Immunology Research Center for Oral and Systemic Health, Beijing Friendship Hospital, Capital Medical University , No. 95 Yong'an Rd, Xicheng District, Beijing 100050 ,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunmei","family":"Zhang","sequence":"additional","affiliation":[{"name":"Salivary Gland Disease Center and Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Beijing 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100069,","place":["China"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guangyong","family":"Sun","sequence":"additional","affiliation":[{"name":"Salivary Gland Disease Center and Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Beijing Laboratory of Oral Health and Beijing Stomatological Hospital, Capital Medical University , No. 9 Fanjiacun Rd, Fengtai District, Beijing 100070 ,","place":["China"]},{"name":"Immunology Research Center for Oral and Systemic Health, Beijing Friendship Hospital, Capital Medical University , No. 95 Yong'an Rd, Xicheng District, Beijing 100050 ,","place":["China"]},{"name":"Medical Research Center, Beijing Institute of Respiratory Medicine and Beijing Chao-Yang Hospital, Capital Medical University , No. 8 Gongti South Rd, Chaoyang District, Beijing 100020 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Gland Disease Center and Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Beijing Laboratory of Oral Health and Beijing Stomatological Hospital, Capital Medical University , No. 9 Fanjiacun Rd, Fengtai District, Beijing 100070 ,","place":["China"]},{"name":"Immunology Research Center for Oral and Systemic Health, Beijing Friendship Hospital, Capital Medical University , No. 95 Yong'an Rd, Xicheng District, Beijing 100050 ,","place":["China"]},{"name":"Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Capital Medical University , No. 10 Xitoutiao, Youanmenwai, Fengtai District, Beijing 100069,","place":["China"]},{"name":"Laboratory for Oral and General Health Integration and Translation, Beijing Tiantan Hospital, Capital Medical University , No. 119 South 4th Ring West Rd, Fengtai District, Beijing 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