Advanced Search

Journal of Traditional Chinese Medicine >

2025 , Vol. 45 >Issue 6: 1273 - 1282

DOI: https://doi.org/10.19852/j.cnki.jtcm.20250314.001

Original Articles

Effect of acupuncture on brain microenvironment in rats with post-stroke limb spasticity based on single-cell transcriptome sequencing technology

  • Qiongshuai ZHANG ,
  • Yi LI ,
  • Fang CAO ,
  • Mujun ZHI ,
  • Le WANG ,
  • Ruyao LIU ,
  • Juanjuan FENG
Expand
  • 1 Department of Rehabilitation, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, China
    2 Department of Acupuncture, the First Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450000, China

Received date: 2025-01-05

  Accepted date: 2025-03-04

  Online published: 2025-03-14

Supported by

Clinical Efficacy and Mechanism of Action of Intermittent Oro-esophageal Tube Feeding in the Treatment of Post-Stroke Dysphagia(LHGJ20220348);Mechanism of Acupuncture in Preventing and Treating Limb Spasticity after Stroke Based on Mechanistic Target of Rapamycin Lactylation and Mitophagy(232300420256)

Fold

Abstract

OBJECTIVE: To investigate the possible mechanisms by which acupuncture improves post-stroke limb spasticity using single-cell sequencing technology.

METHODS: Thirty-two rats were randomly assigned to four groups: Control, Sham, Model, and Acupuncture. The middle cerebral artery occlusion (MCAO) model was established, and the acupuncture groups received acupuncture treatment. After treatment, brain morphological changes and the degree of neurological impairment were assessed. The effect of acupuncture on the proportion of brain cell types in the ischemic penumbra of MCAO rats was analyzed using single-cell transcriptomics, and the expression and enrichment of differentially expressed genes were examined. Finally, selected differential genes were validated by Western blot and quantitative real-time polymerase chain reaction.

RESULTS: Triphenyltetrazolium chloride staining showed that the infarct area in MCAO rats was significantly reduced after acupuncture. Garcia scoring, hematoxylin-eosin staining, Nissl staining, and terminal deoxynucleotidyl transferase dUTP nick end labeling demonstrated that acupuncture reduced brain damage. Enzyme-linked immunosorbent assay results showed that acupuncture significantly decreased serum inflammatory factors, including interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Single-cell transcriptome analysis revealed marked changes in cell type proportions between the Acupuncture and Model groups. A total of 207 differential genes were identified, including 157 upregulated and 50 downregulated genes. Analysis of macrophage-specific differential genes in the ischemic penumbra showed enrichment in Gene Ontology terms such as Ras protein signal transduction and regulation of GTPase activity, and Kyoto Encyclopedia of Genes and Genomes pathways including lysosome, axon guidance, and mitogen-activated protein kinase signaling. S100a8 and leukocyte specific transcript 1 (LST1) were identified as key differential genes.

CONCLUSION: These findings suggest that the key differential genes S100a8 and LST1 may alleviate post-stroke limb spasticity by regulating the inflammatory response in the ischemic penumbra.

Key words： acupuncture; single-cell gene expression analysis; macrophages; post-stroke spasticity; central microenvironment

Cite this article

Qiongshuai ZHANG , Yi LI , Fang CAO , Mujun ZHI , Le WANG , Ruyao LIU , Juanjuan FENG . Effect of acupuncture on brain microenvironment in rats with post-stroke limb spasticity based on single-cell transcriptome sequencing technology[J]. Journal of Traditional Chinese Medicine, 2025 , 45(6) : 1273 -1282 . DOI: 10.19852/j.cnki.jtcm.20250314.001

References

1. Li S, Francisco GE, Rymer WZ. A new definition of poststroke spasticity and the interference of spasticity with motor recovery from acute to chronic stages. Neurorehabil Neural Repair 2021; 35:601.
2. Ghannadi S, Shariat A, Ansari NN, et al. The effect of dry needling on lower limb dysfunction in poststroke survivors. J Stroke Cerebrovasc Dis 2020; 29: 104814.
3. Xie L, Mao G, Xie Y, et al. Efficacy of Baishao Luoshi decoction on synaptic plasticity in rats with post stroke spasticity. J Tradit Chin Med 2023; 43: 295.
4. Gomez-Cuaresma L, Lucena-Anton D, Gonzalez-Medina G, Martin-Vega FJ, Galan-Mercant A, Luque-Moreno C. Effectiveness of stretching in post-stroke spasticity and range of motion: systematic review and Meta-analysis. J Pers Med 2021; 11: 1074.
5. Lim SM, Yoo J, Lee E, et al. Acupuncture for spasticity after stroke: a systematic review and Meta-analysis of randomized controlled trials. Evid Based Complement Alternat Med 2015; 2015: 870398.
6. Xue C, Jiang C, Zhu Y, et al. Effectiveness and safety of acupuncture for post-stroke spasticity: a systematic review and Meta-analysis. Front Neurol 2022; 13: 942597.
7. Zhang QS, Zhang Y, Ji GC, Xu XH, Wang YF, Song BL. Clinical observation on cluster acupuncture at scalp points combined with exercise therapy in treatment of limb spasm after stroke. Zhong Guo Zhen Jiu 2022; 42: 377.
8. Zhang HY, Zhao YJ, Zhang PJ, et al. Acupuncture ameliorates neurological function by suppressing microglia polarization and inflammatory response after cerebral ischemia in rats. Zhen Ci Yan Jiu 2022; 47: 941.
9. Wang WB, Yang LF, He QS, et al. Mechanisms of electroacupuncture effects on acute cerebral ischemia/reperfusion injury: possible association with upregulation of transforming growth factor beta 1. Neural Regen Res 2016; 11: 1099.
10. Zhang Y, Mao X, Lin R, Li Z, Lin J. Electroacupuncture ameliorates cognitive impairment through inhibition of Ca2+-mediated neurotoxicity in a rat model of cerebral ischaemia-reperfusion injury. Acupunct Med 2018; 36: 401.
11. Xie C, Gao X, Luo Y, Pang Y, Li M. Electroacupuncture modulates stromal cell-derived factor-1α expression and mobilization of bone marrow endothelial progenitor cells in focal cerebral ischemia/reperfusion model rats. Brain Res 2016; 1648: 119.
12. Yoo J, Seo JJ, Eom JH, Hwang DY. Effects of stromal cell-derived factor 1α delivered at different phases of transient focal ischemia in rats. Neuroscience 2012; 209: 171.
13. Shi L, Sun Z, Su W, et al. Treg cell-derived osteopontin promotes microglia-mediated white matter repair after ischemic stroke. Immunity 2021; 54: 1527.
14. Nguyen TH, Vicidomini R, Choudhury SD, et al. scRNA-seq data from the larval Drosophila ventral cord provides a resource for studying motor systems function and development. Dev Cell 2024; 59: 1210.
15. Longa EZ, Weinstein PR, Carlson S, Cummins R. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 1989; 20: 84.
16. Wang Y, Tian M, Tan J, et al. Irisin ameliorates neuroinflammation and neuronal apoptosis through integrin αVβ5/AMPK signaling pathway after intracerebral hemorrhage in mice. J Neuroinflammation 2022; 19: 82.
17. Percie DSN, Ahluwalia A, Alam S, et al. Reporting animal research: explanation and elaboration for the ARRIVE guidelines 2.0. PLoS Biol 2020; 18: e3000411.
18. Hopkins AM, DeSimone E, Chwalek K, Kaplan DL. 3D in vitro modeling of the central nervous system. Prog Neurobiol 2015; 125: 1.
19. Hubert T, Grimal S, Carroll P, Fichard-Carroll A. Collagens in the developing and diseased nervous system. Cell Mol Life Sci 2009; 66: 1223.
20. Anlar B, Gunel-Ozcan A. Tenascin-R: role in the central nervous system. Int J Biochem Cell Biol 2012; 44: 1385.
21. Fisher M, Feuerstein G, Howells DW, et al. Update of the stroke therapy academic industry roundtable preclinical recommendations. Stroke 2009; 40: 2244.
22. Wang JH, Zhang TZ, Li XL, et al. Effect of scalp acupuncture stimulation on cerebral infarct volume and expression of IL-10, IL-6 and IL-1β in para-hippocampal gyrus in acute ischemic cerebrovascular disease rats. Zhen Ci Yan Jiu 2019; 44: 405.
23. de Sousa N, Pinho AG, Monteiro S, et al. Acute baclofen administration promotes functional recovery after spinal cord injury. Spine J 2023; 23: 379.
24. Nguyen JN, Chauhan A. Bystanders or not? Microglia and lymphocytes in aging and stroke. Neural Regen Res 2023; 18: 1397.
25. Liu W, Wang X, Yang S, et al. Electroacupunctre improves motor impairment via inhibition of microglia-mediated neuroinflammation in the sensorimotor cortex after ischemic stroke. Life Sci 2016; 151: 313.
26. Gelderblom M, Leypoldt F, Steinbach K, et al. Temporal and spatial dynamics of cerebral immune cell accumulation in stroke. Stroke 2009; 40: 1849.
27. Hu X, Leak RK, Shi Y, et al. Microglial and macrophage polarization — new prospects for brain repair. Nat Rev Neurol 2015; 11: 56.
28. Peng XY, Yuan B, Tian T, et al. Effects of electro-scalp acupuncture on inflammatory response and microglial polarization in the ischemic cortex of rats with ischemic stroke. Zhong Guo Zhen Jiu 2023; 43: 1050.
29. Zhang SH, Wang YL, Zhang CX, et al. Effect of interactive dynamic scalp acupuncture on post-stroke cognitive function, depression, and anxiety: a multicenter, randomized, controlled trial. Chin J Integr Med 2022; 28: 106.
30. Sun X, Wang Y, Zhao Y, et al. Activation of the Epac/Rap 1 signaling pathway alleviates blood-brain barrier disruption and brain damage following cerebral ischemia/reperfusion injury. Int Immunopharmacol 2023; 117: 110014.
31. Takahashi H, Asahina R, Fujioka M, et al. Ras-like Gem GTPase induced by Npas4 promotes activity-dependent neuronal tolerance for ischemic stroke. Proc Natl Acad Sci U S A 2021; 118.
32. Zhang X, Chen C, Ling C, et al. EGFR tyrosine kinase activity and Rab GTPases coordinate EGFR trafficking to regulate macrophage activation in sepsis. Cell Death Dis 2022; 13: 934.
33. Wang S, Song R, Wang Z, Jing Z, Wang S, Ma J. S100A8/A 9 in inflammation. Front Immunol 2018; 9: 1298.
34. Yan L, Bjork P, Butuc R, et al. Beneficial effects of quinoline-3-carboxamide (ABR-215757) on atherosclerotic plaque morphology in S100A12 transgenic ApoE null mice. Atherosclerosis 2013; 228: 69.
35. Morrow DA, Wang Y, Croce K, et al. Myeloid-related protein 8/14 and the risk of cardiovascular death or myocardial infarction after an acute coronary syndrome in the pravastatin or atorvastatin evaluation and infection therapy: thrombolysis in myocardial infarction (PROVE IT-TIMI 22) trial. Am Heart J 2008; 155: 49.
36. Kraakman MJ, Lee MK, Al-Sharea A, et al. Neutrophil-derived S100 calcium-binding proteins A8/A9 promote reticulated thrombocytosis and atherogenesis in diabetes. J Clin Invest 2017; 127: 2133.
37. Song R, Struhl K. S100A8/S100A 9 cytokine acts as a transcriptional coactivator during breast cellular transformation. Sci Adv 2021; 1: 7.
38. Ganta VC, Choi M, Farber CR, Annex BH. Antiangiogenic VEGF(165)b regulates macrophage polarization via S100A8/ S100A9 in peripheral artery disease. Circulation 2019; 139: 226.
39. Pickering TG, Hall JE, Appel LJ, et al. Recommendations for blood pressure measurement in humans: an AHA scientific statement from the Council on High Blood Pressure Research Professional and Public Education Subcommittee. J Clin Hypertens (Greenwich) 2005; 7: 102.
40. Heidemann J, Kebschull M, Tepasse PR, Bettenworth D. Regulated expression of leukocyte-specific transcript (LST) 1 in human intestinal inflammation. Inflamm Res 2014; 63: 513.
41. Fabisik M, Tureckova J, Pavliuchenko N, et al. Regulation of inflammatory response by transmembrane adaptor protein LST1. Front Immunol 2021; 12: 618332.
Options
Outlines

/

〈 〉