{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,29]],"date-time":"2026-10-29T14:26:38Z","timestamp":1793283998782,"version":"4.5.0"},"reference-count":54,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T00:00:00Z","timestamp":1759968000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Endocrinol."],"abstract":"<jats:sec><jats:title>Objective<\/jats:title><jats:p>Anemarrhenae Rhizoma (AR) is a traditional Chinese medicine widely used for the treatment of type 2 diabetes mellitus (T2DM). However, the specific bioactive constituents responsible for its <jats:italic>in vivo<\/jats:italic> effects and their underlying mechanisms of action remain unclear. We hypothesise that serum-absorbed and metabolised AR components modulate key metabolic and inflammatory pathways in T2DM. To test this hypothesis, this study employs an integrated strategy combining metabolomics with serum-urine pharmacochemistry and network pharmacology to systematically identify AR\u2019s active constituents and elucidate their multi-target mechanisms in T2DM management.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>UHPLC-Q-TOF-MS coupled with multivariate statistical analysis was employed to identify the AR-derived constituents in serum and urine of T2DM rats. Network pharmacology was utilised to predict the targets of the AR\u2019s active components, while biochemical assays, liver histopathology, and metabolomics were performed to evaluate its therapeutic effects. Molecular docking and molecular dynamics (MD) simulations were conducted to assess the binding affinities between key components and their targets.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>77 AR components were identified, among which 47 prototypes and 11 metabolites were detected in serum and urine. The key bioactive constituents included sarsasapogenin, markogenin\/neogitogenin, digitogenin, norathyriol, and mangiferin. AR treatment significantly reduced blood glucose and lipid levels, ameliorated insulin resistance, attenuated inflammation, and modulated the PPAR and NF-\u03baB signalling pathways. Serum metabolomics analysis revealed 35 differential metabolites, with linoleic acid metabolism and PPAR signalling identified as the predominant metabolic pathways. Molecular docking and MD simulations demonstrated strong binding affinity between core components and key targets (PPARA, NFKB1, IL6, AKT1, IL1B). Pharmacological validation confirmed AR\u2019s therapeutic efficacy in T2DM through regulation of these core targets.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>AR ameliorates T2DM by suppressing NF-\u03baB signalling and activating PPAR pathways, thereby improving metabolic dysregulation.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fendo.2025.1618584","type":"journal-article","created":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T04:22:02Z","timestamp":1759983722000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":2,"title":["Metabolomics integrated with network pharmacology and serum-urine pharmacochemistry unveils the antidiabetic mechanism of Anemarrhenae Rhizoma"],"prefix":"10.3389","volume":"16","author":[{"given":"Xunlong","family":"Zhong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huaidong","family":"Peng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chang","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunhua","family":"Xiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinyu","family":"Zhu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haixuan","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruolun","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanmei","family":"Zhong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingwen","family":"Feng","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,10,9]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1016\/j.mcna.2014.08.012","article-title":"Oral antihyperglycemic treatment options for type 2 diabetes mellitus","volume":"99","author":"Brietzke","year":"2015","journal-title":"Med Clin North Am"},{"key":"B2","doi-asserted-by":"publisher","first-page":"592","DOI":"10.1080\/13880209.2024.2374794","article-title":"Evidence of traditional Chinese medicine for treating type 2 diabetes mellitus: from molecular mechanisms to clinical efficacy","volume":"62","author":"Ni","year":"2024","journal-title":"Pharm Biol"},{"key":"B3","doi-asserted-by":"publisher","first-page":"42","DOI":"10.1016\/j.jep.2014.02.013","article-title":"The genus Anemarrhena Bunge: A Review on ethnopharmacology, phytochemistry and pharmacology","volume":"153","author":"Wang","year":"2014","journal-title":"J Ethnopharmacol"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpba.2023.115386","article-title":"Lipidomics of the erythrocyte membrane and network pharmacology to explore the mechanism of mangiferin from Anemarrhenae Rhizoma in treating type 2 diabetes mellitus rats","volume":"230","author":"Zhong","year":"2023","journal-title":"J Pharm BioMed Anal"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.1016\/j.phymed.2024.155439","article-title":"Integrated serum pharmacochemistry and network pharmacology to explore the mechanism of Yi-Shan-Hong formula in alleviating chronic liver injury","volume":"128","author":"Zhao","year":"2024","journal-title":"Phytomedicine"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.1002\/pca.3120","article-title":"Integrated multicomponent analysis based on UHPLC-Q-Exactive Orbitrap-MS and network pharmacology to elucidate the potential mechanism of Baoyuan decoction against idiopathic pulmonary fibrosis","volume":"33","author":"Zhang","year":"2022","journal-title":"Phytochem Anal"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1016\/j.lfs.2023.122347","article-title":"Integrated non-targeted metabolomics and network pharmacology to reveal the mechanisms of berberine in the long-term treatment of PTZ-induced epilepsy","volume":"336","author":"Zhu","year":"2024","journal-title":"Life Sci"},{"key":"B8","doi-asserted-by":"publisher","DOI":"10.2174\/0113816128304628240526071425","article-title":"The potential mechanism of eriodictyol in treating Alzheimer\u2019s Disease: a Study on computer-assisted investigational strategies","volume":"30","author":"Du","year":"2024","journal-title":"Curr Pharm Des"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.1016\/j.arabjc.2023.105486","article-title":"Integration of pharmacodynamics, metabolomics and network pharmacology to elucidate the effect of Prunella vulgaris seed oil in the treatment of hyperlipidemia","volume":"17","author":"Zeng","year":"2024","journal-title":"Arab J Chem"},{"key":"B10","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2023.117340","article-title":"Integrated network pharmacology and metabolomics to investigate the effects and possible mechanisms of dehydroevodiamine against ethanol-induced gastric ulcers","volume":"319","author":"Wang","year":"2024","journal-title":"J Ethnopharmacol"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2015.06.015","article-title":"Metabolites characterization of timosaponin AIII in vivo and in vitro by using liquid chromatography-mass spectrometry","volume":"997","author":"Sun","year":"2015","journal-title":"J Chromatogr B"},{"key":"B12","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1016\/j.intimp.2015.02.016","article-title":"Timosaponin AIII and its metabolite sarsasapogenin ameliorate colitis in mice by inhibiting NF-\u03baB and MAPK activation and restoring Th17\/Treg cell balance","volume":"25","author":"Lim","year":"2015","journal-title":"Int Immunopharmacol"},{"key":"B13","doi-asserted-by":"publisher","DOI":"10.1038\/s41401-020-0427-1","article-title":"Sarsasapogenin improves adipose tissue inflammation and ameliorates insulin resistance in high-fat diet-fed C57bl\/6j mice","volume":"42","author":"Yu","year":"2021","journal-title":"Acta Pharmacol Sin"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1016\/j.phymed.2021.153686","article-title":"Sarsasapogenin attenuates Alzheimer-like encephalopathy in diabetes","volume":"91","author":"Zhang","year":"2021","journal-title":"Phytomedicine"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jafc.8b03763","article-title":"Absorption, metabolism, and pharmacokinetics profiles of norathyriol, an aglycone of mangiferin, in rats by HPLC-MS\/MS","volume":"66","author":"Guo","year":"2018","journal-title":"J Agric Food Chem"},{"key":"B16","doi-asserted-by":"publisher","first-page":"68","DOI":"10.3109\/13880209.2013.814691","article-title":"Mangiferin and its aglycone, norathyriol, improve glucose metabolism by activation of AMP-activated protein kinase","volume":"52","author":"Wang","year":"2014","journal-title":"Pharm Biol"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.1007\/s00125-014-3315-8","article-title":"Norathyriol reverses obesity- and high-fat-diet-induced insulin resistance in mice through inhibition of PTP1B","volume":"57","author":"Ding","year":"2014","journal-title":"Diabetologia"},{"key":"B18","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2018.00201","article-title":"Mangiferin improves hepatic lipid metabolism mainly through its metabolite-norathyriol by modulating Sirt-1\/Ampk\/Srebp-1c signaling","volume":"9","author":"Li","year":"2018","journal-title":"Front Pharmacol"},{"key":"B19","doi-asserted-by":"publisher","DOI":"10.1016\/j.phymed.2023.154733","article-title":"Jiang-Tang-San-Huang pill alleviates type 2 diabetes mellitus through modulating the gut microbiota and bile acids metabolism","volume":"113","author":"Tawulie","year":"2023","journal-title":"Phytomedicine"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2021.104186","article-title":"Liver proteomics analysis reveals abnormal metabolism of bile acid and arachidonic acid in Chinese hamsters with type 2 diabetes mellitus","volume":"239","author":"Wang","year":"2021","journal-title":"J Proteomics"},{"key":"B21","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.305789","article-title":"Glucose and insulin induction of bile acid synthesis: mechanisms and implication in diabetes and obesity","volume":"287","author":"Li","year":"2012","journal-title":"J Biol Chem"},{"key":"B22","doi-asserted-by":"publisher","first-page":"16","DOI":"10.5604\/01.3001.0010.5672","article-title":"The role of bile acids in glucose metabolism and their relation with diabetes","volume":"16","author":"Gonz\u00e1lez-Regueiro","year":"2017","journal-title":"Ann Hepatol"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.1016\/j.heliyon.2024.e26326","article-title":"Lipidomics based on UHPLC\/Q-TOF-MS to characterize lipid metabolic profiling in patients with newly diagnosed type 2 diabetes mellitus with dyslipidemia","volume":"10","author":"Zhong","year":"2024","journal-title":"Heliyon"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.1016\/j.jpba.2023.115725","article-title":"Serum and urine metabolomics study revealed theamelioration of gynura bicolor extract on high fat diet-fed and streptozotocin-induced type 2 diabetic mice based on UHPLC-MS\/MS","volume":"236","author":"Ding","year":"2023","journal-title":"J Pharm BioMed Anal"},{"key":"B25","doi-asserted-by":"publisher","DOI":"10.1016\/j.fbio.2024.104631","article-title":"Integrated serum and liver metabolomics decipher the hepatoprotective mechanisms of mangiferin sodium salt through modulating alcohol metabolism on alcoholic liver disease","volume":"61","author":"Li","year":"2024","journal-title":"Food Bioscience"},{"key":"B26","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.16239","article-title":"A review on the relationship between Arachidonic acid 15-Lipoxygenase (ALOX15) and diabetes mellitus","volume":"11","author":"He","year":"2023","journal-title":"PeerJ"},{"key":"B27","doi-asserted-by":"publisher","first-page":"465","DOI":"10.1007\/978-3-319-55330-6_25","article-title":"Insulin resistance and mitochondrial dysfunction","volume":"982","author":"Gonzalez-Franquesa","year":"2017","journal-title":"Adv Exp Med Biol"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1210\/er.2014-1100","article-title":"TLR4 at the crossroads of nutrients, gut microbiota, and metabolic inflammation","volume":"36","author":"Velloso","year":"2015","journal-title":"Endocr Rev"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1210\/en.2014-1467","article-title":"Palmitate-induced impairments of \u03b2-cell function are linked with generation of specific ceramide species via acylation of sphingosine","volume":"156","author":"Manukyan","year":"2015","journal-title":"Endocrinology"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1016\/j.biochi.2015.09.006","article-title":"Palmitate activates mTOR\/p70S6K through AMPK inhibition and hypophosphorylation of raptor in skeletal muscle cells: Reversal by oleate is similar to metformin","volume":"118","author":"Kwon","year":"2015","journal-title":"Biochimie"},{"key":"B31","doi-asserted-by":"publisher","DOI":"10.3390\/jcm9082615","article-title":"Oleic acid protects against insulin resistance by regulating the genes related to the PI3K signaling pathway","volume":"9","author":"L\u00f3pez-G\u00f3mez","year":"2020","journal-title":"J Clin Med"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1194\/jlr.M047357","article-title":"Dietary omega-3 fatty acids modulate the eicosanoid profile in man primarily via the CYP-epoxygenase pathway","volume":"55","author":"Fischer","year":"2014","journal-title":"J Lipid Res"},{"key":"B33","doi-asserted-by":"publisher","DOI":"10.1016\/j.jdiacomp.2014.04.008","article-title":"Serum phospholipid omega-3 polyunsaturated fatty acids and insulin resistance in type 2 diabetes mellitus and non-alcoholic fatty liver disease","volume":"28","author":"Lou","year":"2014","journal-title":"J Diabetes Complications"},{"key":"B34","doi-asserted-by":"publisher","DOI":"10.1161\/01.Cir.0000158483.13093.9d","article-title":"Metabolic syndrome: a comprehensive perspective based on interactions between obesity, diabetes, and inflammation","volume":"111","author":"Dandona","year":"2005","journal-title":"Circulation"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.1016\/j.dsx.2020.07.028","article-title":"Effect of augmented glycation in mobilization of plasma free fatty acids in type 2 diabetes mellitus","volume":"14","author":"Chandra","year":"2020","journal-title":"Diabetes Metab Syndr"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1016\/j.nut.2019.05.007","article-title":"Arachidonic acid-rich arasco oil has anti-inflammatory and antidiabetic actions against streptozotocin + high fat diet induced diabetes mellitus in wistar rats","volume":"66","author":"Gundala","year":"2019","journal-title":"Nutrition"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1016\/j.nut.2014.12.028","article-title":"Supplementation of \u03b1-linolenic acid improves serum adiponectin levels and insulin sensitivity in patients with type 2 diabetes","volume":"31","author":"Gomes","year":"2015","journal-title":"Nutrition"},{"key":"B38","article-title":"Docosahexaenoic acid has an anti-diabetic effect in streptozotocin-induced diabetic mice","volume":"7","author":"Li","year":"2014","journal-title":"Int J Clin Exp Med"},{"key":"B39","doi-asserted-by":"publisher","DOI":"10.3390\/ijms221910573","article-title":"The PPARA and PPAR\u03b3 epigenetic landscape in cancer and immune and metabolic disorders","volume":"22","author":"Porcuna","year":"2021","journal-title":"Int J Mol Sci"},{"key":"B40","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-024-07781-7","article-title":"Remodelling of the translatome controls diet and its impact on tumorigenesis","volume":"633","author":"Yang","year":"2024","journal-title":"Nature"},{"key":"B41","doi-asserted-by":"publisher","first-page":"201","DOI":"10.1186\/s12933-020-01172-8","article-title":"Marked effects of novel selective peroxisome proliferator-activated receptor \u03b1 modulator, pemafibrate in severe hypertriglyceridemia: preliminary report","volume":"19","author":"Iitake","year":"2020","journal-title":"Cardiovasc Diabetol"},{"key":"B42","doi-asserted-by":"publisher","DOI":"10.3390\/ijms241612665","article-title":"The role of PIk3R1 in metabolic function and insulin sensitivity","volume":"24","author":"Tsay","year":"2023","journal-title":"Int J Mol Sci"},{"key":"B43","doi-asserted-by":"publisher","DOI":"10.3389\/fphar.2022.842879","article-title":"Co-Crystal of rosiglitazone with berberine ameliorates hyperglycemia and insulin resistance through the PI3k\/Akt\/TXNIP Pathway","volume":"13","author":"He","year":"2022","journal-title":"Vivo vitro. Front Pharmacol"},{"key":"B44","doi-asserted-by":"publisher","DOI":"10.1016\/j.jgg.2021.09.004","article-title":"Functionalized gadofullerene ameliorates impaired glycolipid metabolism in type 2 diabetic mice","volume":"49","author":"Wu","year":"2022","journal-title":"J Genet Genomics"},{"key":"B45","doi-asserted-by":"publisher","DOI":"10.1016\/j.jff.2021.104532","article-title":"Linoleic acid exerts antidiabetic effects by inhibiting protein tyrosine phosphatases associated with insulin resistance","volume":"83","author":"Yoon","year":"2021","journal-title":"J Funct Foods"},{"key":"B46","doi-asserted-by":"publisher","DOI":"10.1016\/j.nut.2018.05.033","article-title":"Phosphorylation of protein kinase B, the key enzyme in insulin-signaling cascade, is enhanced in linoleic and arachidonic acid-treated HT29 and HepG2 cells","volume":"57","author":"Mariniello","year":"2019","journal-title":"Nutrition"},{"key":"B47","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2023.116966","article-title":"Therapeutic effect of hosta plantaginea (Lam.) aschers flowers on acute pharyngitis through inhibition of multi-Inflammatory pathways in rats","volume":"318","author":"Wang","year":"2024","journal-title":"J Ethnopharmacol"},{"key":"B48","doi-asserted-by":"publisher","first-page":"793","DOI":"10.1016\/j.biochi.2010.12.012","article-title":"A mechanistic approach for islet amyloid polypeptide aggregation to develop anti-amyloidogenic agents for type-2 diabetes","volume":"93","author":"Ahmad","year":"2011","journal-title":"Biochimie"},{"key":"B49","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbr.2016.08.047","article-title":"TLR4 is a link between diabetes and Alzheimer\u2019s disease","volume":"316","author":"Huang","year":"2017","journal-title":"Behav Brain Res"},{"key":"B50","doi-asserted-by":"publisher","DOI":"10.1039\/d0fo03051a","article-title":"Sandalwood seed oil ameliorates hepatic insulin resistance by regulating the Jnk\/Nf-\u03bab inflammatory and PI3k\/Akt insulin signaling pathways","volume":"12","author":"Zhang","year":"2021","journal-title":"Food Funct"},{"key":"B51","doi-asserted-by":"publisher","DOI":"10.1002\/dmrr.3735","article-title":"Association of plasma advanced glycation end-products and their soluble receptor with type 2 diabetes among Chinese adults","volume":"40","author":"Chen","year":"2024","journal-title":"Diabetes Metab Res Rev"},{"key":"B52","doi-asserted-by":"publisher","DOI":"10.1016\/j.jep.2021.114713","article-title":"Mulberry leaves ameliorate diabetes via regulating metabolic profiling and AGEs\/RAGE and p38 MAPK\/NF-\u03baB pathway","volume":"283","author":"Li","year":"2022","journal-title":"J Ethnopharmacol"},{"key":"B53","doi-asserted-by":"publisher","first-page":"e3620","DOI":"10.1002\/dmrr.3620","article-title":"Th1 bias of liver mucosal-associated invariant T cells promotes hepatic gluconeogenesis in type 2 diabetes mellitus","volume":"39","author":"Tang","year":"2023","journal-title":"Diabetes Metab Res Rev"},{"key":"B54","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1038\/s41392-024-01757-9","article-title":"NF-\u03baB in biology and targeted therapy: new insights and translational implications","volume":"9","author":"Guo","year":"2024","journal-title":"Signal Transduct Target Ther"}],"container-title":["Frontiers in Endocrinology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2025.1618584\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T04:22:04Z","timestamp":1759983724000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fendo.2025.1618584\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,9]]},"references-count":54,"alternative-id":["10.3389\/fendo.2025.1618584"],"URL":"https:\/\/doi.org\/10.3389\/fendo.2025.1618584","relation":{},"ISSN":["1664-2392"],"issn-type":[{"value":"1664-2392","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,9]]},"article-number":"1618584"}}