The aim of this study was to investigate the effects of acupuncture on a rat model of chronic migraine (CM) and explore the underlying mechanism of action from the perspective of the gut–brain axis.
Methods:
A total of 24 male Sprague-Dawley (SD) rats were randomly allocated into control, model and acupuncture groups (n = 8 each). The CM model was established by intraperitoneal injection of nitroglycerin (NTG). Acupuncture was administered at GV20 and bilateral PC6/LR3/ST36 for rats in the acupuncture group once a day for 9 days. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of 5-hydroxytryptamine (5-HT), calcitonin gene-related peptide (CGRP) and vasoactive intestinal peptide (VIP) in plasma. A combination of 16S rDNA sequencing and liquid chromatography-mass spectrometry (LC-MS) metabolomics was adopted to investigate the role of the gut–brain axis in migraine chronification and the effect of acupuncture on CM.
Results:
Acupuncture treatment significantly attenuated hyperalgesia in CM model rats and regulated serum levels of brain–gut peptides, including 5-HT, CGRP and VIP. Furthermore, the gut microbial community structure and metabolic profile changed in CM rats and acupuncture impacted the changes. Notably, acupuncture modulated 10 gut microbial genera and 13 fecal metabolites.
Conclusion:
These findings suggest that the gut–brain axis may play an important role in the chronification of migraine, and the regulation of gut microbiota and metabolites may be one of the mechanisms underlying the analgesic effect of acupuncture in CM.
Headache Classification Committee of the International Headache Society (IHS). The international classification of headache disorders, 3rd edition. Cephalalgia2018; 38(1): 1–211.
2.
MeleticheDMLoflandJHYoungWB.Quality-of-life differences between patients with episodic and transformed migraine. Headache2001; 41(6): 573–578.
3.
BuseDCManackASerranoD, et al. Sociodemographic and comorbidity profiles of chronic migraine and episodic migraine sufferers. J Neurol Neurosurg Psychiatry2010; 81(4): 428–432.
On behalf of the School of Advanced Studies of the European Headache Federation (EHF-SAS); ArzaniMRazeghi JahromiSGhorbaniZ, et al. Gut–brain axis and migraine headache: a comprehensive review. J Headache Pain2020; 21: 15.
6.
AamodtAHStovnerLJHagenK, et al. Comorbidity of headache and gastrointestinal complaints: the Head-HUNT Study. Cephalalgia2007; 27(12): 1444–1451.
7.
MoissetXBommelaerGBoubeM, et al. Migraine prevalence in inflammatory bowel disease patients: a tertiary-care centre cross-sectional study. Eur J Pain2017; 21(9): 1550–1560.
8.
de RoosNMGiezenaarCGTRoversJMP, et al. The effects of the multispecies probiotic mixture Ecologic®Barrier on migraine: results of an open-label pilot study. Benef Microbes2015; 6(5): 641–646.
9.
MartamiFToghaMSeifishahparM, et al. The effects of a multispecies probiotic supplement on inflammatory markers and episodic and chronic migraine characteristics: a randomized double-blind controlled trial. Cephalalgia2019; 39(7): 841–853.
10.
XieYZhouGXuY, et al. Effects of diet based on IgG elimination combined with probiotics on migraine plus irritable bowel syndrome. Pain Res Manag2019; 2019: 7890461–7890466.
11.
KangLTangWZhangY, et al. The gut microbiome modulates nitroglycerin-induced migraine-related hyperalgesia in mice. Cephalalgia2022; 42(6): 490–499.
12.
TangYLiuSShuH, et al. Gut microbiota dysbiosis enhances migraine-like pain via TNF-α upregulation. Mol Neurobiol2020; 57(1): 461–468.
13.
XuJZhangFQPeiJ, et al. Acupuncture for migraine without aura: a systematic review and meta-analysis. J Integr Med2018; 16(5): 312–321.
14.
XianMShenLZhanS, et al. Integrated 16S rRNA gene sequencing and LC/MS-based metabolomics ascertained synergistic influences of the combination of acupuncture and NaoMaiTong on ischemic stroke. J Ethnopharmacol2022; 293: 115281.
15.
JangJHYeomMJAhnS, et al. Acupuncture inhibits neuroinflammation and gut microbial dysbiosis in a mouse model of Parkinson’s disease. Brain Behav Immun2020; 89: 641–655.
16.
YangBHeMChenX, et al. Acupuncture effect assessment in APP/PS1 transgenic mice: regulation of learning-memory abilities, gut microbiota, and microbial metabolites. Comput Math Methods Med2022; 2022: 1–20.
17.
HongJChenJKanJ, et al. Effects of acupuncture treatment in reducing sleep disorder and gut microbiota alterations in PCPA-induced insomnia mice. Evid Based Complement Alternat Med2020; 2020: 3626120–3626114.
18.
LiuCHanJZhangW, et al. Analgesic effect of Shugan Tiaoshen acupuncture combined with Tuina in treating migraine. J Clin Acupunct Moxibustion2021; 37(3): 20–24.
19.
WangTSunXGuY, et al. Preliminary discussion on soothing liver and regulating mind acupuncture for treatment of mental diseases. World J Integr Tradit West Med2021; 16(9): 1751–1764.
20.
WanKXuQShiY, et al. Electroacupuncture produces analgesic effects via cannabinoid CB1 receptor–mediated GABAergic neuronal inhibition in the rostral ventromedial medulla. Chin Med2025; 20: 30.
21.
AllaisGChiarleGSinigagliaS, et al. Acupuncture treatment of migraine, nausea, and vomiting in pregnancy. Neurol Sci2019; 40(Suppl. 1): 213–215.
22.
LuMJYuZHeY, et al. Electroacupuncture at ST36 modulates gastric motility via vagovagal and sympathetic reflexes in rats. World J Gastroenterol2019; 25(19): 2315–2326.
23.
BaoCHWangCYLiGN, et al. Effect of mild moxibustion on intestinal microbiota and NLRP6 inflammasome signaling in rats with post-inflammatory irritable bowel syndrome. World J Gastroenterol2019; 25(32): 4696–4714.
24.
LiuGHLiuHMChenYS, et al. Effect of electroacupuncture in mice with dextran sulfate sodium–induced colitis and the influence of gut microbiota. Evid Based Complement Alternat Med2020; 2020: 2087903–2087913.
25.
WangLAnJSongS, et al. Electroacupuncture preserves intestinal barrier integrity through modulating the gut microbiota in DSS-induced chronic colitis. Life Sci2020; 261: 118473.
26.
ThapaRMagarATShresthaJ, et al. Influence of gut and lung dysbiosis on lung cancer progression and their modulation as promising therapeutic targets: a comprehensive review. Medcomm (2020)2024; 5(12): e70018.
27.
CurtoMLionettoLNegroA, et al. Altered kynurenine pathway metabolites in serum of chronic migraine patients. J Headache Pain2016; 17: 47.
28.
WishartDS.Metabolomics for investigating physiological and pathophysiological processes. Physiol Rev2019; 99(4): 1819–1875.
29.
WuBXMaJYHuangXC, et al. Acupuncture as a potential therapeutic approach for Tourette syndrome: modulation of neurotransmitter levels and gut microbiota. Chin J Integr Med2025; 31(8): 735–742.
30.
LvZLiuRSuK, et al. Acupuncture ameliorates breast cancer–related fatigue by regulating the gut microbiota–gut–brain axis. Front Endocrinol2022; 13: 921119.
31.
MoyeLSPradhanAAA. Animal model of chronic migraine-associated pain. Curr Protoc Neurosci2017; 80: 9601–9609.
32.
SufkaKJStaszkoSMJohnsonAP, et al. Clinically relevant behavioral endpoints in a recurrent nitroglycerin migraine model in rats. J Headache Pain2016; 17: 40.
33.
China Association for Acupuncture Moxibustion. Nomenclature and location of acupuncture points for laboratory animals—Part 2: rat. Acupunct Res2021; 46(4): 351–352.
34.
Galeano-CastañedaYBascuñánPSerreD, et al. Trans-stadial fate of the gut bacterial microbiota in Anopheles albimanus. Acta Trop2020; 201: 105204.
35.
GromskiPSMuhamadaliHEllisDI, et al. A tutorial review: metabolomics and partial least squares–discriminant analysis—a marriage of convenience or a shotgun wedding. Anal Chim Acta2015; 879: 10–23.
36.
BylesjöMErikssonDSjödinA, et al. Orthogonal projections to latent structures as a strategy for microarray data normalization. BMC Bioinformatics2007; 8: 207.
37.
WangMYuXGengW, et al. Effect of manual acupuncture preconditioning on behavior and serum CGRP, SP, IL-1β and TNF-α levels in migraine rats. Acupunct Res2018; 43(6): 375–379.
38.
LiYLiHHanJ.Study of analgesic and anti-inflammatory mechanisms in prevention and treatment of migraine with acupuncture therapy of soothing liver and regulating spirit method. Chin Arch Tradit Chin Med2018; 36(11): 2643–2646.
39.
PradhanAASmithMLMcGuireB, et al. Characterization of a novel model of chronic migraine. Pain2014; 155(2): 269–274.
40.
WangMYuXGengW, et al. Manual acupuncture stimulation regulates expression of receptor activity-modifying protein 1 and 5-HT1D receptor proteins and genes in migraine rats. Acupunct Res2018; 43(7): 440–444.
41.
Cernuda-MorollónELarrosaDRamónC, et al. Interictal increase of CGRP levels in peripheral blood as a biomarker for chronic migraine. Neurology2013; 81(14): 1191–1196.
42.
Cernuda-MorollónEMartínez-CamblorPAlvarezR, et al. Increased VIP levels in peripheral blood outside migraine attacks as a potential biomarker of cranial parasympathetic activation in chronic migraine. Cephalalgia2015; 35(4): 310–316.
43.
SeyboldVS. The role of peptides in central sensitization. Handb Exp Pharmacol2009(194): 451–491.
44.
OttossonAEdvinssonL.Release of histamine from dural mast cells by substance P and calcitonin gene-related peptide. Cephalalgia1997; 17(3): 166–174.
45.
FerrariMDOdinkJTapparelliC, et al. Serotonin metabolism in migraine. Neurology1989; 39(9): 1239–1242.
46.
NanNGongMXWangQ, et al. Wuzhuyu decoction relieves hyperalgesia by regulating central and peripheral 5-HT in chronic migraine model rats. Phytomedicine2022; 96: 153905.
47.
ChenJWangQWangA, et al. Structural and functional characterization of the gut microbiota in elderly women with migraine. Front Cell Infect Microbiol2020; 9: 470.
48.
WenZHeMPengC, et al. Metabolomics and 16S rRNA gene sequencing analyses of changes in the intestinal flora and biomarkers induced by Gastrodia-Uncaria treatment in a rat model of chronic migraine. Front Pharmacol2019; 10: 1425.
49.
XieLLZhaoYLYangJ, et al. Electroacupuncture prevents osteoarthritis of high-fat diet-induced obese rats. Biomed Res Int2020; 2020: 9380965.
50.
LindenbergFKrychLFieldenJ, et al. Expression of immune regulatory genes correlates with the abundance of specific Clostridiales and Verrucomicrobia species in the equine ileum and cecum. Sci Rep2019; 9: 12674.
51.
TakahashiK.Effect of Enterococcus faecalis 2001 on colitis and depressive-like behavior in dextran sulfate sodium-treated mice: involvement of the brain–gut axis. J Neuroinflammation2019; 16: 16.
52.
FungTCVuongHELunaCDG, et al. Intestinal serotonin and fluoxetine exposure modulate bacterial colonization in the gut. Nat Microbiol2019; 4(12): 2064–2073.
53.
YuJBiXYuB, et al. Isoflavones: anti-inflammatory benefit and possible caveats. Nutrients2016; 8: 361.
54.
ShimosatoT.Oral administration of Flavonifractor plautii, a bacteria increased with green tea consumption, promotes recovery from acute colitis in mice via suppression of IL-17. Front Nutr2021; 7: 9.
55.
DongLZhengQChengY, et al. Gut microbial characteristics of adult patients with epilepsy. Front Neurosci2022; 16: 803538.
56.
WangYOuyangMGaoX, et al. Phocea, Pseudoflavonifractor, and Lactobacillus intestinalis: three potential biomarkers of gut microbiota that affect progression and complications of obesity-induced type 2 diabetes mellitus. Diabetes Metab Syndr Obes2020; 13: 835–850.
57.
LeeCHongSNPaikNY, et al. CD1d modulates colonic inflammation in NOD2−/− mice by altering the intestinal microbial composition comprising Acetatifactor muris. J Crohns Colitis2019; 13: 11.
58.
LabusJSOsadchiyVHsiaoEY, et al. Evidence for an association of gut microbial Clostridia with brain functional connectivity and gastrointestinal sensorimotor function in patients with irritable bowel syndrome, based on tripartite network analysis. Microbiome2019; 7: 45.
59.
RenCLiuJZhouJ, et al. Low levels of serum serotonin and amino acids identified in migraine patients. Biochem Biophys Res Commun2018; 496: 267–273.
PeroutkaSJ. Migraine: a chronic sympathetic nervous system disorder. Headache2004; 44(1): 53–64.
62.
D’AndreaGPeriniFTerrazzinoS, et al. Contributions of biochemistry to the pathogenesis of primary headaches. Neurol Sci2004; 25(Suppl. 3): S89–92.
63.
SugaharaM.4-Aminobutyraldehyde as a precursor convertible to γ-aminobutyric acid in vivo. J Biochem1983; 93: 6.
64.
PeekALLeaverAMFosterS, et al. Increased GABA+ in people with migraine, headache, and pain conditions: a potential marker of pain. J Pain2021; 22(12): 1631–1645.
65.
AguilaMRRebbeckTLeaverAM, et al. The association between clinical characteristics of migraine and brain GABA levels: an exploratory study. J Pain2016; 17(10): 1058–1067.
66.
TillakaratneNJMedina-KauweLGibsonKM.Gamma-aminobutyric acid (GABA) metabolism in mammalian neural and nonneural tissues. Comp Biochem Physiol A Physiol1995; 112(2): 247–263.
67.
YonedaYRobertsEDietzGWJr.A new synaptosomal biosynthetic pathway of glutamate and GABA from ornithine and its negative feedback inhibition by GABA. J Neurochem1982; 38(6): 1686–1694.
68.
KatanoYFujinamiSKawakoshiA, et al. Complete genome sequence of oscillibacter valericigenes Sjm18-20T (=NBRC 101213T). Stand Genomic Sci2012; 6: 406–414.
69.
NaseribafroueiAHestadKAvershinaE, et al. Correlation between the human fecal microbiota and depression. Neurogastroenterol Motil2014; 26(8): 1155–1162.
70.
LarouteVYasaroCNarinW, et al. GABA production in lactococcus lactis is enhanced by arginine and co-addition of malate. Front Microbiol2016; 7: 1050.
71.
JungMYLeeCSeoMJ, et al. Characterization of a potential probiotic bacterium Lactococcus raffinolactis WiKim0068 isolated from fermented vegetable using genomic and in vitro analyses. BMC Microbiol2020; 20: 136.
72.
ProuskyJSeelyD.The treatment of migraines and tension-type headaches with intravenous and oral niacin (nicotinic acid): systematic review of the literature. Nutr J2005; 4: 3.
