Objective  To systematically explore the mechanism of astragaloside IV (AS-IV) in the treatment of ischemic stroke (IS) using integrated pharmacology method.

Methods  Potential targets of AS-IV and disease targets of IS were retrieved from multiple databases. The common targets were screened using the Venny website. A protein-protein interaction (PPI) network was constructed via the STRING database, and core targets were analyzed using Cytoscape 3.9.1 software with the CytoHubba plugin. The common targets of AS-IV and IS were imported into the DAVID database for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis to identify relevant pathways. AutoDock Vina 1.2.5 software was used to validate the molecular docking between AS-IV and the core targets, and Pymol 2.5 software was used to visualize the docking results. An oxygen-glucose deprivation/reoxygenation (OGD/R) model was established using in vitro cultured human neuroblastoma SH-SY5Y cells to verify the mechanism of AS-IV in treating IS.

Results  A total of 371 potential targets for AS-IV, 121 targets related to IS, and 41 intersection targets were screened. PPI network analysis identified key core targets such as BCL2, IL-6, and TNF. GO and KEGG analyses indicated that the mechanism of AS-IV in treating IS was associated with the NF-κB signaling pathway. Molecular docking results demonstrated a strong binding ability between AS-IV and the 10 core targets. Cell experiment results showed that compared with the blank group, the levels of TNF, NF-κB, IL-6, Caspase-3, and IL-1β in the model group were significantly increased, while the BCL2 level was significantly decreased (P<0.05). Compared with the model group, the levels of TNF, NF-κB, IL-6, Caspase-3, and IL-1β in the AS-IV low-, medium-, and high-dose groups, as well as the TNF inhibitor group, were significantly decreased, while BCL2 levels were significantly increased (P<0.05). Compared with the AS-IV medium-dose group, the levels of TNF, NF-κB, IL-6, Caspase-3, and IL-1β in the AS-IV medium-dose+TNF agonist group were significantly increased, and the BCL2 level was significantly decreased (P<0.05).

Conclusion  AS-IV may inhibit inflammation by regulating the TNF/NF-κB signaling pathway, thereby exerting its therapeutic effect on IS.

1.Feigin VL, Brainin M, Norrving B, et al. World Stroke Organization (WSO): Global Stroke Fact Sheet 2022[J]. Int J Stroke, 2022, 17(1): 18-29. DOI: 10.1177/17474930211065917.

2.Alsbrook DL, Di Napoli M, Bhatia K, et al. Neuroinflammation in acute ischemic and hemorrhagic stroke[J]. Curr Neurol Neurosci Rep, 2023, 23(8): 407-431. DOI: 10.1007/s11910-023-01282-2.

3.Neves D, Salazar IL, Almeida RD, et al. Molecular mechanisms of ischemia and glutamate excitotoxicity[J]. Life Sciences, 2023, 121814. DOI: 10.1016/j.lfs.2023.121814.

4.Zong P, Feng J, Li CX, et al. Activation of endothelial TRPM2 exacerbates blood-brain barrier degradation in ischemic stroke[J]. Cardiovasc Res, 2024, 120(2): 188-202. DOI: 10.1093/cvr/cvad126.

5.Powers WJ, Rabinstein AA, Ackerson T, et al. Guidelines for the early management of patients with acute ischemic stroke: 2019 update to the 2018 guidelines for the early management of acute ischemic stroke: a guideline for healthcare professionals from the American Heart Association/American Stroke Association[J]. Stroke, 2019, 50(12): e344-e418. DOI: 10.1161/STR.0000000000000211.

6.Campbell BCV, De Silva DA, Macleod MR, et al. Ischaemic stroke[J]. Nat Rev Dis Primers, 2019, 5(1): 70. DOI: 10.1038/s41572-019-0118-8.

7.Song T, Zhang Y, Zhu L, et al. The role of JAK/STAT signaling pathway in cerebral ischemia-reperfusion injury and the therapeutic effect of traditional Chinese medicine: a narrative review[J]. Medicine (Baltimore), 2023, 102(46): e35890. DOI: 10.1097/md.0000000000035890.

8.诸葛陆杰, 方燕, 金华倩, 等. 补阳还五汤上调 miR-199a-5p 表达促进脑缺血大鼠神经发生和血管生成[J]. 浙江大学学报（医学版）, 2020, 49(6): 687-696. [Zhuge LJ, Fang Y, Jin HQ, et al. Chinese medicine Buyang Huanwu decoction promotes neurogenesis and angiogenesis in ischemic stroke rats by upregulating miR-199a-5p expression[J]. Journal of Zhejiang University (Medical Sciences), 2020, 49(6): 687-696.] DOI: 10.3785/j.issn.1008-9292.2020.12.03.

9.Chen C, Bu X, Deng L, et al. Astragaloside IV as a promising therapeutic agent for liver diseases: current landscape and future perspectives[J]. Front Pharmacol, 2025, 16: 1574154. DOI: 10.3389/fphar.2025.1574154.

10.Yao M, Zhang L, Wang L. Astragaloside IV: a promising natural neuroprotective agent for neurological disorders[J]. Biomed Pharmacother, 2023, 159: 114229. DOI: 10.1016/j.biopha.2023.114229.

11.Zhang W, Yang Y, Zhang X, et al. Astragaloside IV relieves central sensitization by regulating astrocytic ROS/NF-κB nuclear translocation signaling in chronic migraine male rats[J]. Phytother Res, 2025, 39(3): 1438-1452. DOI: 10.1002/ptr.8436.

12.Liang Y, Chen B, Liang D, et al. Pharmacological effects of astragaloside IV: a review[J]. Molecules, 2023, 28(16): 6118. DOI: 10.3390/molecules28166118.

13.Su X, Guo H, Zhou Y, et al. Astragaloside IV attenuates high glucose-induced NF-κB-mediated inflammation through activation of PI3K/AKT-ERK-dependent Nrf2/ARE signaling pathway in glomerular mesangial cells[J]. Phytother Res, 2023, 37(9): 4133-4148. DOI: 10.1002/ptr.7875.

14.Xiao L, Dai Z, Tang W, et al. Astragaloside IV alleviates cerebral ischemia-reperfusion injury through NLRP3 inflammasome-mediated pyroptosis inhibition via activating Nrf2[J]. Oxid Med Cell Longev, 2021, 2021: 9925561. DOI: 10.1155/2021/9925561.

15.Shi YH, Zhang XL, Ying PJ, et al. Neuroprotective effect of astragaloside IV on cerebral ischemia/reperfusion injury rats through sirt1/mapt pathway[J]. Front Pharmacol, 2021, 12: 639898. DOI: 10.3389/fphar.2021.639898.

16.Ding Y, Jie K, Xin L, Shao B. Astragaloside IV plays a neuroprotective role by promoting PPARγ in cerebral ischemia-reperfusion rats[J]. Behav Brain Res, 2025, 476: 115267. DOI: 10.1016/j.bbr.2024.115267.

17.张苗苗, 胡利明, 周全, 等. 人参皂苷Ro对氧糖剥夺/复糖复氧诱导SH-SY5Y细胞损伤的保护作用及机制[J]. 中国神经免疫学和神经病学杂志, 2025, 32(1): 35-41. [Zhang MM, Hu LM, Zhou Q, et al. Therapeutic effect and mechanism of ginsenoside Ro on SH-SY5Y cell injury induced by oxygen-glucose deprivation/reoxygenation[J]. Chinese Journal of Neuroimmunology and Neurology, 2025, 32(1): 35-41.] DOI: 10.3969/j.issn.1006-2963.2025.01.006.

18.Xu Z, Yang D, Huang X, et al. Astragaloside IV protects 6-hydroxydopamine-induced SH-SY5Y cell model of Parkinson's disease via activating the JAK2/STAT3 pathway[J]. Front Neurosci, 2021, 15: 631501. DOI: 10.3389/fnins.2021.631501.

19.Liu S, Wang Y, Wen H, et al. Hydroxysafflor yellow A inhibits TNF-α-induced inflammation of human fetal lung fibroblasts via NF-κB signaling pathway[J]. Evid Based Complement Alternat Med, 2019, 2019: 4050327. DOI: 10.1155/2019/4050327.

20.Dos Passos Junior RR, de Freitas RA, Reppetti J, et al. High levels of tumor necrosis factor-alpha reduce placental aquaporin 3 expression and impair in vitro trophoblastic cell migration[J]. Front Physiol, 2021, 12: 696495. DOI: 10.3389/fphys.2021.696495.

21.Amruta N, Rahman AA, Pinteaux E, et al. Neuroinflammation and fibrosis in stroke: the good, the bad and the ugly[J]. J Neuroimmunol, 2020, 346: 577318. DOI: 10.1016/j.jneuroim. 2020.577318.

22.Lu W, Chen Z, Wen J. Flavonoids and ischemic stroke-induced neuroinflammation: focus on the glial cells[J]. Biomed Pharmacother, 2024, 170: 115847. DOI: 10.1016/j.biopha.2023.115847.

23.Wang D, Wang Y, Shi J, et al. Edaravone dexborneol alleviates ischemic injury and neuroinflammation by modulating microglial and astrocyte polarization while inhibiting leukocyte infiltration[J]. Int Immunopharmacol, 2024, 130: 111700. DOI: 10.1016/j.intimp. 2024.111700.

24.Tirandi A, Sgura C, Carbone F, et al. Inflammatory biomarkers of ischemic stroke[J]. Intern Emerg Med, 2023, 18(3): 723-732. DOI: 10.1007/s11739-023-03201-2.

25.Jayaraj RL, Azimullah S, Beiram R, et al. Neuroinflammation: friend and foe for ischemic stroke[J]. J Neuroinflammation, 2019, 16(1): 142. DOI: 10.1186/s12974-019-1516-2.

26.Mokhtari Sangdehi SR, Hajizadeh Moghaddam A, Ranjbar M. Anti-apoptotic effect of silymarin-loaded chitosan nanoparticles on hippocampal Caspase-3 and BCL-2 expression following cerebral ischemia/reperfusion injury[J]. Int J Neurosci, 2022, 132(11): 1102-1109. DOI: 10.1080/00207454.2020.1860971.

27.Hussar P. Apoptosis regulators BCL-2 and caspase-3[J]. Encyclopedia, 2022, 2(4): 1624-1636. DOI: 10.3390/encyclopedia2040111.

28.Glaviano A, Foo ASC, Lam HY, et al. PI3K/AKT/mTOR signaling transduction pathway and targeted therapies in cancer[J]. Mol Cancer, 2023, 22(1): 138. DOI: 10.1186/s12943-023-01827-6.

29.Barnabei L, Laplantine E, Mbongo W, et al. NF-κB: at the borders of autoimmunity and inflammation[J]. Front Immunol, 2021, 12: 716469. DOI: 10.3389/fimmu.2021.716469.

30.Liu D, Zhong Z, Karin M. NF-κB: a double-edged sword controlling inflammation[J]. Biomedicines, 2022, 10(6): 1250. DOI: 10.3390/biomedicines10061250.

31.Anilkumar S, Wright-Jin E. NF-κB as an inducible regulator of inflammation in the central nervous system[J]. Cells, 2024, 13(6): 485. DOI: 10.3390/cells13060485.

32.Zhang L, Ludden CM, Cullen AJ, et al. Nuclear factor kappa B expression in non-small cell lung cancer[J]. Biomed Pharmacother, 2023, 167: 115459. DOI: 10.1016/j.biopha.2023.115459.

33.Zamanian MY, Golmohammadi M, Gardanova ZR, et al. The roles of neuroinflammation in l-DOPA-induced dyskinesia: dissecting the roles of NF-κB and TNF-α for novel pharmacological therapeutic approaches[J]. Eur J Neurosci, 2025, 61(5): e70034. DOI: 10.1111/ejn.70034.

34.Li H, Yao C, Shi K, et al. Astragaloside IV attenuates hypoxia/reoxygenation injury-induced apoptosis of type II alveolar epithelial cells through miR-21-5p[J]. Bioengineered, 2021, 12(1): 7747-7754. DOI: 10.1080/21655979.2021.1982845.

35.Hou Y, Yan Z, Wan H, et al. A combination of astragaloside IV and hydroxysafflor yellow A attenuates cerebral ischemia-reperfusion injury via NF-κB/NLRP3/caspase-1/GSDMD pathway[J]. Brain Sci, 2024, 14(8): 781. DOI: 10.3390/brainsci14080781.

36.Zhang C, Shi Z, Xu Q, et al. Astragaloside IV alleviates stroke-triggered early brain injury by modulating neuroinflammation and ferroptosis via the Nrf2/HO-1 signaling pathway[J]. Acta Cir Bras, 2023, 38: e380723. DOI: 10.1590/acb380723.

37.Shi G, Chen J, Zhang C, et al. Astragaloside IV promotes cerebral angiogenesis and neurological recovery after focal ischemic stroke in mice via activating PI3K/Akt/mTOR signaling pathway[J]. Heliyon, 2023, 9(12): e22800. DOI: 10.1016/j.heliyon.2023.e22800.