{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T06:17:28Z","timestamp":1775024248761,"version":"3.50.1"},"reference-count":43,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T00:00:00Z","timestamp":1774569600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Natural Science Foundation of Fujian Province of China","award":["2021J01910"],"award-info":[{"award-number":["2021J01910"]}]},{"name":"Natural Science Foundation of Fujian Province of China","award":["2023J01313"],"award-info":[{"award-number":["2023J01313"]}]},{"name":"Open Project Fund of Fujian Key Laboratory of Drug Target Discovery and Structural and Functional Research","award":["FKLDSR-202301"],"award-info":[{"award-number":["FKLDSR-202301"]}]},{"name":"Joint Funds for the innovation of Science and Technology\uff0cFujian province","award":["2025Y9274"],"award-info":[{"award-number":["2025Y9274"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceuticals"],"abstract":"<jats:p>Background\/Objectives: Salidroside, a bioactive phenylethanol glycoside primarily derived from Rhodiola rosea, and its major in vivo metabolite tyrosol exhibit diverse pharmacological activities. However, their direct molecular targets remain poorly defined. Methods: In the present study, an integrated strategy combining transcriptomic profiling, Connectivity Map (CMap) analysis, and multi-level experimental validation was employed. Transcriptomic signatures derived from A549 cells treated with salidroside or tyrosol were queried against the CMap database. Molecular docking, surface plasmon resonance (SPR), and cellular thermal shift assays (CETSA) were performed to predict and validate binding interactions. Functional validation was performed in SH-SY5Y cells. The phosphorylation level of extracellular signal-regulated kinase (ERK), a downstream signaling event of dopamine D2 receptor (DRD2), was detected after salidroside and tyrosol treatment. DRD2 antagonist sulpiride pre-intervention and small interfering RNA (siRNA)-mediated DRD2 knockdown were conducted to verify the receptor dependence of the compounds\u2019 effects. Results: CMap analysis revealed that the transcriptomic signatures of salidroside and tyrosol showed significant similarity to known DRD2 modulators. Molecular docking predicted potential binding interactions between the two compounds and DRD2, which was confirmed by SPR and CETSA to be direct physical binding. Functional studies showed that both compounds rapidly induced DRD2 downstream ERK phosphorylation in SH-SY5Y cells; this effect was abrogated by sulpiride or DRD2 knockdown, indicating DRD2-dependent signaling activation. Conclusions: These findings identify DRD2 as a direct molecular target of salidroside and tyrosol and provide mechanistic insight into their dopaminergic regulatory effects. This study highlights the utility of CMap-guided target discovery combined with rigorous experimental validation for elucidating the molecular mechanisms of natural products.<\/jats:p>","DOI":"10.3390\/ph19040540","type":"journal-article","created":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T14:33:33Z","timestamp":1774622013000},"page":"540","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification of Dopamine D2 Receptor as a Direct Target of Salidroside and Tyrosol by Integrated Transcriptomic and Biophysical Approaches"],"prefix":"10.3390","volume":"19","author":[{"given":"Jizhou","family":"Zhang","sequence":"first","affiliation":[{"name":"Institute of Materia Medica, Fujian Academy of Chinese Medical Science, Fuzhou 350003, China"},{"name":"Key Laboratory of Fujian Province Universities on Ion Channel and Signal Transduction in Cardiovascular Diseases, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350108, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8855-2173","authenticated-orcid":false,"given":"Kan","family":"Lin","sequence":"additional","affiliation":[{"name":"Fujian Medical University Union Hospital, Fuzhou 350001, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chang","family":"Jiang","sequence":"additional","affiliation":[{"name":"Institute of Materia Medica, Fujian Academy of Chinese Medical Science, Fuzhou 350003, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jiabing","family":"Zheng","sequence":"additional","affiliation":[{"name":"Fujian Medical University Union Hospital, Fuzhou 350001, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huihui","family":"Huang","sequence":"additional","affiliation":[{"name":"Fujian Key Laboratory of Drug Target Discovery and Structural and Functional Research, Fujian Medical University, Fuzhou 350108, China"},{"name":"Department of Pharmacology, School of Pharmacy, Fujian Medical University, Fuzhou 350108, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7142-1991","authenticated-orcid":false,"given":"Jing","family":"Han","sequence":"additional","affiliation":[{"name":"Institute of Materia Medica, Fujian Academy of Chinese Medical Science, Fuzhou 350003, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,3,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Guo, N., Zhu, M., Han, X., Sui, D., Wang, Y., and Yang, Q. (2014). The Metabolism of Salidroside to Its Aglycone P-Tyrosol in Rats Following the Administration of Salidroside. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0103648"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"512","DOI":"10.2174\/1389200221666200610172105","article-title":"Salidroside\u2014Can It Be a Multifunctional Drug?","volume":"21","author":"Magani","year":"2020","journal-title":"Curr. Drug Metab."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"109268","DOI":"10.1016\/j.cbi.2020.109268","article-title":"Salidroside: A Review of Its Recent Advances in Synthetic Pathways and Pharmacological Properties","volume":"339","author":"Zhang","year":"2021","journal-title":"Chem. Biol. Interact."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1016\/j.foodchem.2013.04.004","article-title":"Tyrosol Exerts a Protective Effect against Dopaminergic Neuronal Cell Death in in Vitro Model of Parkinson\u2019s Disease","volume":"141","author":"Dewapriya","year":"2013","journal-title":"Food Chem."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Karkovi\u0107 Markovi\u0107, A., Tori\u0107, J., Barbari\u0107, M., and Jakobu\u0161i\u0107 Brala, C. (2019). Hydroxytyrosol, Tyrosol and Derivatives and Their Potential Effects on Human Health. Molecules, 24.","DOI":"10.3390\/molecules24102001"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"110458","DOI":"10.1016\/j.biopha.2020.110458","article-title":"Salidroside as a Potential Neuroprotective Agent for Ischemic Stroke: A Review of Sources, Pharmacokinetics, Mechanism and Safety","volume":"129","author":"Fan","year":"2020","journal-title":"Biomed. Pharmacother."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1694","DOI":"10.21037\/atm-21-5752","article-title":"Salidroside Inhibits NLRP3 Inflammasome Activation and Apoptosis in Microglia Induced by Cerebral Ischemia\/Reperfusion Injury by Inhibiting the TLR4\/NF-\u03baB Signaling Pathway","volume":"9","author":"Liu","year":"2021","journal-title":"Ann. Transl. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1016\/j.fct.2019.01.039","article-title":"Modulation of LPS-Induced Nitric Oxide Production in Intestinal Cells by Hydroxytyrosol and Tyrosol Metabolites: Insight into the Mechanism of Action","volume":"125","author":"Serreli","year":"2019","journal-title":"Food Chem. Toxicol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Lammi, C., Bellumori, M., Cecchi, L., Bartolomei, M., Bollati, C., Clodoveo, M.L., Corbo, F., Arnoldi, A., and Mulinacci, N. (2020). Extra Virgin Olive Oil Phenol Extracts Exert Hypocholesterolemic Effects through the Modulation of the LDLR Pathway: In Vitro and Cellular Mechanism of Action Elucidation. Nutrients, 12.","DOI":"10.3390\/nu12061723"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1080\/07391102.2019.1575283","article-title":"Exploration of Interaction Mechanism of Tyrosol as a Potent Anti-Inflammatory Agent","volume":"38","author":"Yadav","year":"2020","journal-title":"J. Biomol. Struct. Dyn."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1989","DOI":"10.4103\/1673-5374.172317","article-title":"Neuroprotective Effects of Salidroside on Focal Cerebral Ischemia\/Reperfusion Injury Involves the Nuclear Erythroid 2-Related Factor 2 Pathway","volume":"10","author":"Han","year":"2015","journal-title":"Neural Regen. Res."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Zheng, J., Zhang, J., Han, J., Zhao, Z.C., and Lin, K. (2024). The Effect of Salidroside in Promoting Endogenous Neural Regeneration after Cerebral Ischemia\/Reperfusion Involves Notch Signaling Pathway and Neurotrophic Factors. BMC Complement. Med. Ther., 24.","DOI":"10.1186\/s12906-024-04597-w"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Zhong, Z.-F., Han, J., Zhang, J.-Z., Xiao, Q., Chen, J.-Y., Zhang, K., Hu, J., and Chen, L.-D. (2019). Neuroprotective Effects of Salidroside on Cerebral Ischemia\/Reperfusion-Induced Behavioral Impairment Involves the Dopaminergic System. Front. Pharmacol., 10.","DOI":"10.3389\/fphar.2019.01433"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1038\/nrc2044","article-title":"The Connectivity Map: A New Tool for Biomedical Research","volume":"7","author":"Lamb","year":"2007","journal-title":"Nat. Rev. Cancer"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1289","DOI":"10.1016\/j.drudis.2012.07.017","article-title":"Applications of Connectivity Map in Drug Discovery and Development","volume":"17","author":"Qu","year":"2012","journal-title":"Drug Discov. Today"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Song, J. (2025). Applications of the Cellular Thermal Shift Assay to Drug Discovery in Natural Products: A Review. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26093940"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"D901","DOI":"10.1093\/nar\/gkm958","article-title":"DrugBank: A Knowledgebase for Drugs, Drug Actions and Drug Targets","volume":"36","author":"Wishart","year":"2008","journal-title":"Nucleic Acids Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1002\/bab.1719","article-title":"Salidroside Protected against MPP+-Induced Parkinson\u2019s Disease in PC12 Cells by Inhibiting Inflammation, Oxidative Stress and Cell Apoptosis","volume":"66","author":"Zhou","year":"2019","journal-title":"Biotechnol. Appl. Biochem."},{"key":"ref_19","first-page":"529","article-title":"Salidroside Protects SH-SY5Y from Pathogenic \u03b1-Synuclein by Promoting Cell Autophagy via Mediation of mTOR\/p70S6K Signaling","volume":"20","author":"Chen","year":"2019","journal-title":"Mol. Med. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"108833","DOI":"10.1016\/j.pharmthera.2025.108833","article-title":"Recent Advances in Target Identification Technology of Natural Products","volume":"269","author":"Liu","year":"2025","journal-title":"Pharmacol. Ther."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Jiang, H., Hu, C., and Chen, M. (2021). The Advantages of Connectivity Map Applied in Traditional Chinese Medicine. Front. Pharmacol., 12.","DOI":"10.3389\/fphar.2021.474267"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zhu, Y., Yang, H., Han, L., Mervin, L.H., Hosseini-Gerami, L., Li, P., Wright, P., Trapotsi, M.-A., Liu, K., and Fan, T.-P. (2023). In Silico Prediction and Biological Assessment of Novel Angiogenesis Modulators from Traditional Chinese Medicine. Front. Pharmacol., 14.","DOI":"10.3389\/fphar.2023.1116081"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1055\/a-1948-3179","article-title":"The Therapeutic Potential of Salidroside for Parkinson\u2019s Disease","volume":"89","author":"Li","year":"2023","journal-title":"Planta Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1038\/nature25758","article-title":"Structure of the D2 Dopamine Receptor Bound to the Atypical Antipsychotic Drug Risperidone","volume":"555","author":"Wang","year":"2018","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e1900044","DOI":"10.1002\/minf.201900044","article-title":"Probing Binding Landscapes and Molecular Recognition Mechanisms of Atypical Antipsychotic Drugs towards the Selective Targeting of D2 Dopamine Receptor","volume":"38","author":"Olotu","year":"2019","journal-title":"Mol. Inform."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1111\/j.1471-4159.2008.05831.x","article-title":"L-DOPA Activates ERK Signaling and Phosphorylates Histone H3 in the Striatonigral Medium Spiny Neurons of Hemiparkinsonian Mice","volume":"108","author":"Santini","year":"2009","journal-title":"J. Neurochem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e1473242024","DOI":"10.1523\/JNEUROSCI.1473-24.2024","article-title":"Optogenetic Control of Dopamine Receptor 2 Reveals a Novel Aspect of Dopaminergic Neurotransmission in Motor Function","volume":"45","author":"Kim","year":"2025","journal-title":"J. Neurosci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"105989","DOI":"10.1016\/j.phrs.2021.105989","article-title":"Salvianolic Acid A Relieves Cognitive Disorder after Chronic Cerebral Ischemia: Involvement of Drd2\/Cryab\/NF-\u03baB Pathway","volume":"175","author":"Yang","year":"2022","journal-title":"Pharmacol. Res."},{"key":"ref_29","first-page":"4137","article-title":"Salidroside Prevents Tumor Necrosis Factor-\u03b1-induced Vascular Inflammation by Blocking Mitogen-activated Protein Kinase and NF-\u03baB Signaling Activation","volume":"18","author":"Li","year":"2019","journal-title":"Exp. Ther. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1989","DOI":"10.2147\/DDDT.S136792","article-title":"Salidroside Pretreatment Attenuates Apoptosis and Autophagy during Hepatic Ischemia-Reperfusion Injury by Inhibiting the Mitogen-Activated Protein Kinase Pathway in Mice","volume":"11","author":"Feng","year":"2017","journal-title":"Drug Des. Dev. Ther."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5204","DOI":"10.1111\/febs.16921","article-title":"Comparative study of the molecular mechanisms underlying the G protein and \u03b2-arrestin-dependent pathways that lead to ERKs activation upon stimulation by dopamine D2 receptor","volume":"290","author":"Liu","year":"2023","journal-title":"FEBS J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"dev202842","DOI":"10.1242\/dev.202842","article-title":"Common Modes of ERK Induction Resolve into Context-Specific Signalling via Emergent Networks and Cell-Type-Specific Transcriptional Repression","volume":"151","author":"Perera","year":"2024","journal-title":"Development"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.neuroscience.2014.01.025","article-title":"The Role of D2-Autoreceptors in Regulating Dopamine Neuron Activity and Transmission","volume":"282","author":"Ford","year":"2014","journal-title":"Neuroscience"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1080\/03602532.2016.1179754","article-title":"Metabolic Disposition and Biological Significance of Simple Phenols of Dietary Origin: Hydroxytyrosol and Tyrosol","volume":"48","author":"Boronat","year":"2016","journal-title":"Drug Metab. Rev."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1124\/pr.117.013995","article-title":"Modulating Dopamine Signaling and Behavior with Chemogenetics: Concepts, Progress, and Challenges","volume":"71","author":"Runegaard","year":"2019","journal-title":"Pharmacol. Rev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"167","DOI":"10.1016\/j.tins.2006.01.002","article-title":"Dopamine in Neurotoxicity and Neuroprotection: What Do D2 Receptors Have to Do with It?","volume":"29","author":"Bozzi","year":"2006","journal-title":"Trends Neurosci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2203","DOI":"10.1152\/jn.00161.2013","article-title":"Supersensitive Presynaptic Dopamine D2 Receptor Inhibition of the Striatopallidal Projection in Nigrostriatal Dopamine-Deficient Mice","volume":"110","author":"Wei","year":"2013","journal-title":"J. Neurophysiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.bbi.2012.11.017","article-title":"Loss of Substance P and Inflammation Precede Delayed Neurodegeneration in the Substantia Nigra after Cerebral Ischemia","volume":"29","author":"Blackabey","year":"2013","journal-title":"Brain Behav. Immun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1124\/pr.110.002642","article-title":"The Physiology, Signaling, and Pharmacology of Dopamine Receptors","volume":"63","author":"Beaulieu","year":"2011","journal-title":"Pharmacol. Rev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"118349","DOI":"10.1016\/j.lfs.2020.118349","article-title":"Post-Ischemic Administration of Dopamine D2 Receptor Agonist Reduces Cell Death by Activating Mitochondrial Pathway following Ischemic Stroke","volume":"259","author":"Kaushik","year":"2020","journal-title":"Life Sci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"6643","DOI":"10.1038\/s41467-024-50964-z","article-title":"A neurodevelopmental disorder mutation locks G proteins in the transitory pre-activated state","volume":"15","author":"Knight","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1437","DOI":"10.1016\/j.cell.2017.10.049","article-title":"A Next Generation Connectivity Map: L1000 Platform and the First 1,000,000 Profiles","volume":"171","author":"Subramanian","year":"2017","journal-title":"Cell"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1124\/pr.118.016790","article-title":"Biased Receptor Signaling in Drug Discovery","volume":"71","author":"Kenakin","year":"2019","journal-title":"Pharmacol. Rev."}],"container-title":["Pharmaceuticals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/4\/540\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,1]],"date-time":"2026-04-01T04:18:29Z","timestamp":1775017109000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8247\/19\/4\/540"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,27]]},"references-count":43,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["ph19040540"],"URL":"https:\/\/doi.org\/10.3390\/ph19040540","relation":{},"ISSN":["1424-8247"],"issn-type":[{"value":"1424-8247","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,27]]}}}