Astragalus injection alleviates diabetic cognitive impairment (DCI) through dual effects of anti-neuroinflammation and antioxidative stress based on network pharmacology analysis and experimental validation

Abstract

Objective: To explore the effects of astragalus injection (AI) in patients with diabetic cognitive impairment (DCI). Methods: First, to screen for effective chemical components of astragalus, we searched Traditional Chinese Medicines Systems Pharmacology (TCMSP) and Analysis Platform databases. We then predicted the corresponding potential target genes using Swiss Target Prediction, searched and filtered three disease databases to identify DCI targets, and constructed a drug-component-target-disease interaction network and protein–protein interaction (PPI) network. The core targets and pathways of AI against DCI were predicted using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analyses. In vivo experiments demonstrated the efficacy of AI in alleviating diabetes-associated cognitive impairment, whereas in vitro studies verified its anti-inflammatory and antioxidant stress mechanisms. Finally, western blot analysis identified key regulatory proteins involved in inflammation and oxidative stress, proposing a potential mechanism by which AI mitigates DCI through modulation of these proteins. Results: We screened 25 active components of astragalus and 251 drug targets and obtained 220 common targets after intersecting 8,036 DCI targets. GO enrichment analysis showed that astragalus mainly treats DCI by regulating the response to external stimuli, lipopolysaccharide, hypoxia, and oxygen levels. KEGG pathway enrichment analysis revealed that it was mainly involved in cancer, calcium signaling, p53 signaling, vascular endothelial growth factor (VEGF) signaling pathway, and Toll-like receptor (TLR) signaling pathways. Experimental results demonstrated that AI alleviates diabetes-associated cognitive impairment through dual mechanisms: activating the Kelch-like ECH-associated protein (Keap1)/nuclear factor-erythroid-derived related factor 2 (Nrf2) signaling pathway to exert antioxidant effects, and inhibiting the TLR-4/nuclear factor-kappa B (NF-κB) signaling pathway to mitigate neuroinflammation, thereby improving cognitive dysfunction and hyperglycemia. Conclusions: Network pharmacology identified potential AI interventions for DCI, and experiments were conducted to confirm some of the major targets in the predictive signaling pathway. Our study offers new insights into the mechanisms of AI treatment for DCI.

Keywords

Astragalus injection / Astragalus membranaceus Cognitive impairment of diabetes / anti-neuroinflammation / antioxidative stress / Network pharmacology

References

[1] Liu Z, Wang H, Yang Z, et al. Causal associations between type 1 diabetes mellitus and cardiovascular diseases: a Mendelian randomization study. Cardiovasc Diabetol 2023; 22(1): 236.

[2] Li W, Huang E, Gao S. Type 1 diabetes mellitus and cognitive impairments: a systematic review. J Alzheimers Dis 2017; 57(1): 29-36.

[3] Biessels GJ, Despa F. Cognitive decline and dementia in diabetes mellitus: mechanisms and clinical implications. Nat Rev Endocrinol 2018; 14(10): 591-604.

[4] Rawlings AM, Sharrett AR, Albert MS, et al. The association of late—life diabetes status and hyperglycemia with incident mild cognitive impairment and dementia: the ARIC study. Diabetes Care 2019; 42(7): 1248-1254.

[5] Roberts RO, Knopman DS, Geda YE, et al. Association of diabetes with amnestic and nonamnestic mild cognitive impairment. Alzheimers Dement 2014; 10(1): 18-26.

[6] Velayudhan L, Poppe M, Archer N, et al. Risk of developing dementia in people with diabetes and mild cognitive impairment. Br J Psychiatry 2010; 196(1): 36-40.

[7] Pickering J, Wong R, Al—Salami H, et al. Cognitive deficits in type—1 diabetes: aspects of glucose, cerebrovascular and amyloid involvement. Pharm Res 2021; 38(9): 1477-1484.

[8] Zhang BL, Dong ED, Liu L, et al. Ten recommendations for the high—quality development of traditional Chinese medicine in the new era. Acupunct Herb Med 2025; 5(2): 131-133.

[9] Pang B, Chen YB, Lyu HT, et al. Modern technological advances in traditional medicine: global insights from the 2025 top 50 high—impact researches. Acupunct Herb Med 2025; 5(4): 491-497.

[10] Cui K, Zhang S, Jiang X, et al. Novel synergic antidiabetic effects of Astragalus polysaccharides combined with Crataegus flavonoids via improvement of islet function and liver metabolism . Mol Med Rep 2016; 13(6): 4737-4744.

[11] Luo BY, Wang WT, Li YW, et al. Tetramethylpyrazine attenuates chronic intermittent hypoxia—exacerbated diabetic atherosclerosis: A mechanistic study of the IRE1α—XBP1 signaling pathway. Acupunct Herb Med 2025; 5(2): 160-172.

[12] Yue SJ, Liu J, Feng W—W, et al. System pharmacology—based dissection of the synergistic mechanism of Huangqi and Huanglian for diabetes mellitus. Front Pharmacol 2017; 8: 694.

[13] Zhang Y, Yuan Y, Zhang J, et al. Astragaloside IV supplementation attenuates cognitive impairment by inhibiting neuroinflammation and oxidative stress in type 2 diabetic mice. Front Aging Neurosci 2022; 14: 1004557.

[14] Wang X, Wang Y, Hu JP, et al. Astragaloside IV, a natural PPARgamma agonist, reduces a beta production in Alzheimer’s disease through inhibition of BACE1. Mol Neurobiol 2017; 54(4): 2939-2949.

[15] Shang L, Wang Y, Li J, et al. Mechanism of Sijunzi Decoction in the treatment of colorectal cancer based on network pharmacology and experimental validation. J Ethnopharmacol 2023; 302(Pt A): 115876.

[16] Furman BL. Streptozotocin—induced diabetic models in mice and rats. Curr Protoc 2021; 1(4): e78.

[17] Janhavi P, Divyashree S, Sanjailal KP, et al. DoseCal: a virtual calculator for dosage conversion between human and different animal species. Arch Physiol Biochem 2022; 128(2): 426-430.

[18] Vorhees CV, Williams MT. Morris water maze: procedures for assessing spatial and related forms of learning and memory. Nat Protoc 2006; 1(2): 848-858.

[19] Sun K, Ding M, Fu C, et al. Effects of dietary wild bitter melon (Momordica charantia var. abbreviate Ser.) extract on glucose and lipid metabolism in HFD/STZ—induced type 2 diabetic rats. J Ethnopharmacol 2023; 306: 116154.

[20] Li X, Gao Y, Li B, et al. Integrated proteomics and metabolomics analysis of D—pinitol function during hippocampal damage in streptozocin—induced aging—accelerated mice. Front Mol Neurosci 2023; 16: 1251513.

[21] Zhu X, Li N, Lv D, et al. Mechanism of Astragali Radix intervention in peritoneal fibrosis based on network pharmacology and molecular docking technology. Drugs Clinic 2022; 37(4): 700-707.

[22] Bettio L, Rajendran L, Gil—Mohapel J. The effects of aging in the hippocampus and cognitive decline. Neurosci Biobehav Rev 2017; 79: 66-86.

[23] Mehta BK, Banerjee S. Minocycline reverses diabetes—associated cognitive impairment in rats. Pharmacol Rep 2019; 71(4): 713-720.

[24] Liu YW, Zhu X, Zhang L, et al. Cerebroprotective effects of ibuprofen on diabetic encephalopathy in rats. Pharmacol Biochem Behav 2014; 117: 128-136.

[25] Hua Y, Xue W, Zhang M, et al. Metabonomics study on the hepatoprotective effect of polysaccharides from different preparations of Angelica sinensis. J Ethnopharmacol 2014; 151(3): 1090-1099.

[26] Liu DS, Zhou YH, Liang ES, et al. Neuroprotective effects of the Chinese Yi—Qi—Bu—Shen recipe extract on injury of rat hippocampal neurons induced by hypoxia/reoxygenation. J Ethnopharmacol 2013; 145(1): 168-174.

[27] Sancar G, Liu S, Gasser E, et al. FGF1 and insulin control lipolysis by convergent pathways. Cell Metab 2022; 34(1): 171-183.e6.

[28] Zhang Y, Gu Y, Ren H, et al. Gut microbiome—related effects of berberine and probiotics on type 2 diabetes (the PREMOTE study). Nat Commun 2020; 11(1): 5015.

[29] Albazal A, Delshad AA, Roghani M. Melatonin reverses cognitive deficits in streptozotocin—induced type 1 diabetes in the rat through attenuation of oxidative stress and inflammation. J Chem Neuroanat 2021; 112: 101902.

[30] Xiang X, Wang X, Wu YM, et al. Activation of GPR55 attenuates cognitive impairment, oxidative stress, neuroinflammation, and synaptic dysfunction in a streptozotocin—induced Alzheimer’s mouse model. Pharmacol Biochem Behav 2022; 214: 173340.

[31] Vittal RH, Waseem SB, Iannucci J, et al. Thrombin signaling contributes to high glucose—induced injury of human brain microvascular endothelial cells. J Alzheimers Dis 2021; 79(1): 211-224.

[32] Zhang Z, Yang H, Yang J, et al. Pseudoginsenoside—F11 attenuates cognitive impairment by ameliorating oxidative stress and neuroinflammation in d‑galactose—treated mice. Int Immunopharmacol 2019; 67: 78-86.

[33] Piatkowska—Chmiel I, Herbet M, Gawronska—Grzywacz M, et al. The role of molecular and inflammatory indicators in the assessment of cognitive dysfunction in a mouse model of diabetes. Int J Mol Sci 2021; 22(8): 3878.

[34] Sun Y, Huang J, Chen Y, et al. Direct inhibition of Keap1—Nrf2 Protein—Protein interaction as a potential therapeutic strategy for Alzheimer’s disease. Bioorg Chem 2020; 103: 104172.

[35] Medhat E, Rashed L, Abdelgwad M, et al. Exercise enhances the effectiveness of vitamin D therapy in rats with Alzheimer’s disease: emphasis on oxidative stress and inflammation. Metab Brain Dis 2020; 35(1): 111-120.

[36] Wardyn JD, Ponsford AH, Sanderson CM. Dissecting molecular cross—talk between Nrf2 and NF—κB response pathways. Biochem Soc Trans 2015; 43(4): 621-626.

[37] Wang T, Zhang X, Li JJ. The role of NF—kappaB in the regulation of cell stress responses. Int Immunopharmacol 2002; 2(11): 1509-1520.

[38] Shi Y, Tu Z, Tang D, et al. The inhibition of LPS—induced production of inflammatory cytokines by HSP70 involves inactivation of the NF—kappaB pathway but not the MAPK pathways. Shock 2006; 26(3): 277-284.

[39] Liu Y, Wu H, Nie Y—C, et al. Naringin attenuates acute lung injury in LPS—treated mice by inhibiting NF—kappaB pathway. Int Immunopharmacol 2011; 11(10): 1606-1612.

[40] Khayatan D, Razavi SM, Arab ZN, et al. Protective effects of curcumin against traumatic brain injury. Biomed Pharmacother 2022; 154: 113621.

[41] Xiong T, Zheng X, Zhang K, et al. Ganluyin ameliorates DSS—induced ulcerative colitis by inhibiting the enteric—origin LPS/TLR4/NF—kappaB pathway. J Ethnopharmacol 2022; 289: 115001.

[42] Wang YM, Ji R, Chen W—W, et al. Paclitaxel alleviated sepsis—induced acute lung injury by activating MUC1 and suppressing TLR—4/NF—kappaB pathway. Drug Des Devel Ther 2019; 13: 3391-3404.

[43] Abdel Baky NA, Al—Najjar AH, Elariny HA, et al. Pramipexole and lactoferrin ameliorate cyclophosphamide—induced haemorrhagic cystitis via targeting Sphk1/S1P/MAPK, TLR—4/NF—kappaB, and NLRP3/caspase—1/IL—1beta signalling pathways and modulating the Nrf2/HO—1 pathway . Int Immunopharmacol 2022; 112: 109282.