Objective  To investigate whether xanthohumol exerts an antianxiety effect by inhibiting FKBP51 in the brain.

Methods  Forty male C57BL/6J mice were randomly divided into five groups, a control group (Con), a model group (Saline), and xanthohumol groups (5, 10, and 20 mg/kg). Except for the Con group, mice in the other groups were given different doses of xanthohumol after inducing anxiety-like behavior using a restraint stress model. The Saline group and Con group were treated with an equal volume of physiological saline via intraperitoneal injection, administered continuously for 7 days. The open-field test and elevated plus-maze test were used to detect anxiety-like behavior in the mice. Western Blot analysis was employed to measure brain-derived neurotrophic factor (BDNF) protein expression levels in the hippocampus of the mice. Golgi staining was used to assess dendritic spine density in mouse hippocampal neurons. Another forty male C57BL/6J mice were used to evaluate the effects on anxiety-like behavior following the stereotactic injection of an adeno-associated virus (AAV) designed to suppress FKBP51. Molecular docking and dynamics simulations were performed to analyze the binding affinity and stability of xanthohumol and FKBP51.

Results  Xanthohumol significantly improved the anxiety-like behavior of restraint-stressed mice in the open field test and elevated plus-maze test, as evidenced by increased exercise time and distance in the central area, as well as an increase in the number and duration of entries into the open arms. Further research showed that xanthohumol upregulated the protein expression level of BDNF in the hippocampus of mice and increased the density of dendritic spines in neurons in the hippocampal CA1 area, thereby promoting neuronal plasticity. Additionally, the expression of FKBP51 was inhibited by stereotactic injection of AAV, and it was found that the anti-anxiety effect of xanthohumol was dependent on the function of FKBP51. Molecular docking and dynamics simulation results indicated that xanthohumol and the FKBP51 protein had a high binding affinity, forming a stable complex structure.

Conclusion  Xanthohumol plays an anti-anxiety effect by inhibiting the FKBP51, promoting BDNF expression, and improving neuronal plasticity, providing a theoretical basis and potential targets for the development of new anti-anxiety drugs.

1.Ren L, Fan Y, Wu W, et al. Anxiety disorders: treatments, models, and circuitry mechanisms[J]. Eur J Pharmacol, 2024, 983: 176994. DOI: 10.1016/j.ejphar.2024.176994.

2.Liu X, Liu H, Wu X, et al. Xiaoyaosan against depression through suppressing LPS mediated TLR4/NLRP3 signaling pathway in "microbiota-gut-brain" axis[J]. J Ethnopharmacol, 2024, 335: 118683. DOI: 10.1016/j.jep.2024.118683.

3.Ahmad A, Khan S, Ali S, et al. Network pharmacology combined with molecular docking and experimental verification to elucidate the effect of flavan-3-ols and aromatic resin on anxiety[J]. Sci Rep, 2024, 14(1): 9799. DOI: 10.1038/s41598-024-58877-z.

4.Doeselaar L, Abromeiit A, Stark T, et al. FKBP51 in glutamatergic forebrain neurons promotes early life stress inoculation in female mice[J]. Nat Commun, 2025, 16(1): 2529. DOI: 10.1038/s41467-025-57952-x.

5.Gao W, Chen P, Hsu H, et al. Xanthohumol, a prenylated chalcone, regulates lipid metabolism by modulating the LXR/RXR-ANGPTL3-LPL axis in hepatic cell lines and high-fat diet-fed zebrafish models[J]. Biomed Pharmacother, 2024, 174: 116598. DOI: 10.1016/j.biopha.2024.116598.

6.Carbone K, Gervasi F. An updated review of the genus humulus: a valuable source of bioactive compounds for health and disease prevention[J]. Plants (Basel), 2022, 11(24): 3434. DOI: 10.3390/plants11243434.

7.Dlugosz A, Blaszak B, Czarnecki D, et al. Mechanism of action and therapeutic potential of xanthohumol in prevention of selected neurodegenerative diseases[J]. Molecules, 2025, 30(3): 694. DOI: 10.3390/molecules30030694.

8.Ma S, Chong Y, Zhang R, et al. Glycyrrhizic acid treatment ameliorates anxiety-like behaviour via GLT1 and Per1/2-dependent pathways[J]. J Ethnopharmacol 2024, 328: 118013. DOI: 10.1016/j.jep.2024.118013.

9.Ma S, Guo X, Han R, et al. Elucidation of the mechanism of action of ailanthone in the treatment of colorectal cancer: integration of network pharmacology, bioinformatics analysis and experimental validation[J]. Front Pharmacol, 2024, 15: 1355644. DOI: 10.3389/fphar.2024.1355644.

10.Babter A, Scottk K, Vos T, et al. Global prevalence of anxiety disorders: a systematic review and meta-regression[J]. Psychol Med, 43(5): 897-910. DOI: 10.1017/S003329171200147X.

11.边明真, 谭涛, 李华南, 等. 基于中西医临床病证特点的广泛性焦虑症动物模型分析[J].天津中医药, 2024, 41(9): 1214-1220. [Bian MZ, Tan T, Li HN, et al. Analysis of animal model of generalized anxiety disorder based on clinical characteristics of TCM and western medicine[J]. Tianjin Journal of Traditional Chinese Medicine, 2024, 41(9): 1214-1220.] DOI: 10.11656/j.issn.1672-1519.2024.09.23.

12.李雪, 李爽, 武盈吉, 等. 啤酒花的研究进展[J]. 吉林医药学院学报, 2019, 40(2): 143-145. [Li X, Li S, Wu YJ, et al. Research progress of Humulus lupulus[J]. Journal of Jilin Medical University, 2019, 40(2): 143-145.] DOI: 10.13845/j.cnki.issn1673-2995.2019.02.025.

13.林柳悦, 蒋益萍, 张巧艳, 等. 啤酒花化学成分和药理活性研究进展[J]. 中国中药杂志, 2017, 42(10): 1830-1836. [Lin  LY, Jiang YP, Zhang QY, et al. Research progress of chemical constituents and pharmacological activities in Humulus lupulus[J]. China Journal of Chinese Materia Medica, 2017, 42(10): 1830-1836.] DOI: 10.19540/j.cnki.cjcmm.20170224.019.

14.Ma S, Zhang R, Li L, et al. Xanthohumol protect cognitive performance in diabetic model rats by inhibiting protein kinase B/nuclear factor kappa-B pathway[J]. Neuroreport, 2025, 32(8): 651-658. DOI: 10.1097/WNR.0000000000001595.

15.Bathina S, Das U. Brain-derived neurotrophic factor and its clinical implications[J]. Arch Med Sci, 2015, 11(6): 1164-1178. DOI: 10.5114/aoms.2015.56342.

16.李晓雷, 王秋妍, 刘洋, 等. 头针联合通督治郁针法对脑卒中后抑郁（痰瘀互结证）患者抑郁症状及血清脑源性神经营养因子、神经生长因子水平的影响[J]. 海南医学, 2025, 36(6): 794-799. [Li XL, Wang QY, Liu Y, et al. Effects of scalpacupuncture combined with Tongdu Zhiyu acupuncture on depressive symptoms and serum BDNF and NGF levels in patients of post-stroke depression with phlegm-stasis mutual aggregation syndrome[J]. Hainan Medical Journal, 2025, 36(6): 794-799.] DOI: 10.3969/j.issn.1003-6350.2025.06.007.

17.王晓歌, 鲍金宇, 杨帅, 等. 跑台运动上调BDNF/TrkB-CREB通路改善神经性疼痛模型大鼠焦虑样行为[J]. 中国实验动物学报, 2024, 32(9): 1149-1159. [Wang XG, Bao JY, Yang S, et al. Treadmill exercise up-regulates BDNF/TrkB-CREB pathway to improve anxiety-like behavior in neuropathic pain rats[J]. Acta Laboratorium Animalis Scientia Sinica, 2024, 32(9): 1149-1159.] DOI: 10.3969/j.issn.1005-4847.2024.09.006.

18.Kovarova V, Bordes J, Mitra S, et al. Deep phenotyping reveals CRH and FKBP51-dependent behavioral profiles following chronic social stress exposure in male mice[J]. Neuropsychopharmacology, 2025, 50(3): 556-567. DOI: 10.1038/s41386-024-02008-9.

19.Jin W. Regulation of BDNF-TrkB signaling and potential therapeutic strategies for parkinson's disease[J]. J Clin Med, 2020, 9(1): 257. DOI: 10.3390/jcm9010257.

20.Anderzhanova E, Hafner K, Genewsky A, et al. The stress susceptibility factor FKBP51 controls S-ketamine-evoked release of mBDNF in the prefrontal cortex of mice[J]. Neurobiol Stress, 2020, 13: 100239. DOI: 10.1016/j.ynstr.2020.100239.