Objective: To investigate the effect of Astragaloside IV (AS-Ⅳ) on the expression of DNA methyltransferase to up-regulate the expression level of vascular endothelial growth factor A (VEGFA) to reduce brain injury caused by acute hypoxia. Methods: Mice hippocampal neurons (HT22 cells) were divided into Control group, Hypoxia group and Astragaloside IV group (H+AS-Ⅳ). HT22 cells were hypoxic for 13 h and reoxygenated for 6 h to establish a hypoxic model. Group H+AS-Ⅳ was treated with 100 μmol/L astragaloside IV at the same time of reoxygenation. The morphological changes of cells were observed by fluorescence inverted microscope. Real-time PCR was used to detect the expression changes of DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3A (DNMT3A), DNA methyltransferase 3B (DNMT3B) and VEGFA at mRNA level. The expression of DNMT1, DNMT3A and DNMT3B at the protein level was detected by Western blot. Results: Compared with the control group, the cell body of HT22 cells became smaller and the antennae became shorter after hypoxia. The mRNA and protein levels of DNMT1 and DNMT3A were decreased (P＜0.05), while the expression of DNMT3B was increased (P＜0.05), and the mRNA level of VEGFA was decreased (P＜0.05). Compared with the hypoxic group, the cell morphology was improved after AS-Ⅳ treatment. The mRNA level and protein level of DNMT1 and DNMT3A were unchanged, while the mRNA level of DNMT3B was decreased (P＜0.05), and the mrna level of VEGFA was increased (P＜0.05). Conclusion: Astragaloside Ⅳ can downregulate the expression of DNMT3B after hypoxia, exerting a neuroprotective effect, and this protective mechanism may be exerted by upregulating the expression of VEGFA.
YANG Haixia
,
JIN Qingling
,
LIN Jiajing
,
XIE Wei
,
BAO Mulan
,
BADE Rengui
. Astragaloside Ⅳ regulates the expression of VEGFA to protect brain from hypoxic through DNA methylation[J]. Journal of Baotou Medical College, 2023
, 39(6)
: 5
-9
.
DOI: 10.16833/j.cnki.jbmc.2023.06.002
[1] Burtscher J, Mallet RT, Burtscher M, et al. Hypoxia and brain aging: Neurodegeneration or neuroprotection?[J]. Ageing Research Reviews, 2021, 68: 101343.
[2] Nyakas C, Buwalda B, Luiten PG. Hypoxia and brain development[J]. Prog Neurobiol, 1996,49(1):1-51.
[3] Cheng S, Zhang X, Feng Q, et al. Astragaloside Ⅳ exerts angiogenesis and cardioprotection after myocardial infarction via regulating PTEN/PI3K/Akt signaling pathway[J]. Life Sciences, 2019, 227: 82-93.
[4] Li L, Gan H, Jin H, et al. Astragaloside Ⅳ promotes microglia/macrophages M2 polarization and enhances neurogenesis and angiogenesis through PPARγ pathway after cerebral ischemia/reperfusion injury in rats[J]. International Immunopharmacology, 2021, 92: 107335.
[5] Yu NK, Baek SH, Kaang BK. DNA methylation-mediated control of learning and memory[J]. Molecular Brain, 2011, 4(1): 5.
[6] Shi G, Zhou X, Wang X, et al. Signatures of altered DNA methylation gene expression after central and peripheral nerve injury[J]. Journal of Cellular Physiology, 2020, 235(6): 5171-5181.
[7] Basang Z, Zhang S, Yang L, et al. Correlation of DNA methylation patterns to the phenotypic features of Tibetan elite alpinists in extreme hypoxia[J]. Journal of Genetics and Genomics, 2021, 48(10): 928-935.
[8] Apte RS, Chen DS, Ferrara N. VEGF in signaling and disease: beyond discovery and development[J]. Cell, 2019, 176(6): 1248-1264.
[9] Mackenzie F, Ruhrberg C. Diverse roles for VEGF-A in the nervous system[J]. Development, 2012, 139(8): 1371-1380.
[10] Zhang N, Chen J, Ferraro GB, et al. Anti-VEGF treatment improves neurological function in tumors of the nervous system[J]. Experimental Neurology, 2018, 299: 326-333.
[11] Choudhry H, Harris AL. Advances in hypoxia-inducible factor biology[J]. Cell Metabolism, 2018, 27(2): 281-298.
[12] Jha NK, Jha SK, Sharma R, et al. Hypoxia-induced signaling activation in neurodegenerative diseases: Targets for new therapeutic Strategies[J]. Journal of Alzheimer's Disease, 2018, 62(1): 15-38.
[13] Semenza GL. HIF-1 and human disease: one highly involved factor[J]. Genes & Development, 2000, 14(16): 1983-1991.
[14] Hartley I, Elkhoury FF, Heon Shin J, et al. Long-lasting changes in DNA methylation following short-term hypoxic exposure in primary hippocampal neuronal cultures[J]. PLoS ONE, 2013, 8(10): e77859.
[15] Lv Y, Zhang C, Jian H, et al. Regulating DNA methylation could reduce neuronal ischemia response and apoptosis after ischemia-reperfusion injury[J]. Gene, 2022, 837: 146689.
[16] Xu S, Li Y, Chen JP, et al. Oxygen glucose deprivation/re-oxygenation-induced neuronal cell death is associated with Lnc-D63785 m6A methylation and miR-422a accumulation[J]. Cell Death & Disease, 2020, 11(9): 816.
[17] Li H, Wang P, Huang F, et al. Astragaloside Ⅳ protects blood-brain barrier integrity from LPS-induced disruption via activating Nrf2 antioxidant signaling pathway in mice[J]. Toxicology and Applied Pharmacology, 2018, 340: 58-66.
[18] Li L, Hou X, Xu R, et al. Research review on the pharmacological effects of astragaloside Ⅳ[J]. Fundamental & Clinical Pharmacology, 2017, 31(1): 17-36.
[19] Zhang J, Wu C, Gao L, et al. Astragaloside Ⅳ derived from Astragalus membranaceus: A research review on the pharmacological effects[J] Adv Pharmacology, 2020,87: 89-112.
[20] Li S, Dou B, Shu S, et al. Suppressing NK cells by astragaloside Ⅳ protects against acute ischemic stroke in mice via inhibiting STAT3[J]. Front Pharmacology, 2021, 12: 802047.
[21] Khaibullina AA, Rosenstein JM, Krum JM. Vascular endothelial growth factor promotes neurite maturation in primary CNS neuronal cultures[J]. Developmental Brain Research, 2004, 148(1): 59-68.
[22] Nanduri J, Semenza GL, Prabhakar NR. Epigenetic changes by DNA methylation in chronic and intermittent hypoxia[J]. American Journal of Physiology-Lung Cellular and Molecular Physiology, 2017, 313(6): L1096-L1100.
[23] Li X, Liu B, Yang J, et al. DNA methylation in promoter region of immune related genes STAT3 and VEGFA and biochemical parameters change in muscle of Japanese flounder under acute hypoxia[J]. Developmental & Comparative Immunology, 2022, 129: 104295.
