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Single-Nucleus RNA-Seq Reveals Neuroprotective Effects of Acupuncture in Chronic Migraine Through Modulation of Glial
Shiqi Sun1, Shuangyuan Hu1, Mingsheng Sun1
1Acupuncture and Tuina School, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan, China.
Background:
Chronic migraine (CM) is a debilitating neurological disorder with limited treatment options. Although acupuncture has demonstrated clinical efficacy in relieving CM symptoms, its cellular and molecular mechanisms remain poorly understood.
Objective:
This study aimed to delineate the cell-type-specific transcriptional landscape and intercellular communication network reshaped by acupuncture.
Methods:
A rat model of CM was induced by repeated administration of nitroglycerin. Acupuncture was applied at GB8 and GB34 points. Single-nucleus RNA sequencing (snRNA-seq) was performed on the TNC region to profile transcriptomic changes across different cell types. Differential expression analysis, functional enrichment, and pseudotime trajectory inference were performed, along with intercellular communication analysis using CellChat.
Results:
Acupuncture alleviated pain-related behaviors and restored aberrant cell compositions in the TNC, especially among neurons, astrocytes, and microglia. It reversed the CM-induced upregulation of inflammatory and oxidative stress-related genes (e.g., Nfkbia, S100a8, S100a9, Penk). The imbalance between neuronal excitability and metabolism was rectified through the modulation of glutamatergic transmission and oxidative phosphorylation pathways. Microglial polarization shifted from pro-inflammatory (M1/M5) to reparative ones (M2/M4), while astrocytic subtypes rebalanced toward anti-inflammatory and metabolic repair states. CellChat analysis showed that acupuncture also remodeled neuron-glia communication disrupted by CM.
Conclusions:
This study sheds light on the cellular and molecular mechanisms by which acupuncture alleviates CM, providing novel insights into its effects on oxidative stress, inflammation, and neuronal function in the TNC. These findings have important implications for both basic science and clinical practice, supporting the potential of acupuncture as a non-invasive, adjunctive therapy for migraine treatment.