To observe the effect of liver-soothing and mental-activity-regulating (LSMAR) needling on the expression of receptor activity-modifying protein 1 (RAMP 1, receptor of calcitonin gene-related peptide), 5-hydroxytryptamine 1 D receptor(5-HT 1 DR) in the spinal trigeminal nucleus (STN) and midbrain in migraine rats, so as to explore its underlying mechanism in relieving migraine.A total of 40 male Wistar rats were randomly divided into control, model, LSMAR and conventional needling groups (n=10 rats in each). The migraine model was established by subcutaneous injection of nitroglycerin at the posterior neck. LSMAR was applied to "Baihui" (GV 20), bilateral "Fengchi" (GB 20), "Neiguan" (PC 6) and "Taichong" (LR 3) in the LSMAR group and conventional needling was applied to "Baihui" (GV 20) and bilateral "Fengchi" (GB 20) in the conventional acupuncture group for 30 min, once a day for 8 days before modeling. The expression levels of RAMP 1 and 5-HT 1 DR proteins and mRNAs in the STN and mesencephalon were detected by real-time fluorescent quantitative PCR and Western blot, separately.Compared with the control group, the expression levels of RAMP 1 protein and mRNA in STN and mesencephalon were significantly increased (P<0.05) and those of 5-HT 1 DR protein and mRNA considerably decreased (P<0.05) in the model group. After the acupuncture treatment, the increased levels of RAMP 1 protein and mRNA and the decreased levels of 5-HT 1 DR protein and mRNA in the STN and midbrain were obviously reversed in the LSMAR and conventional needling groups relevant to the model group (P<0.05). The effect of LSMAR needling was significantly superior to that of conventional needling in down-regulating the expression levels of RAMP 1 mRNA and protein in the STN and mesencephalon (P<0.05) and in up-regulating the expression levels of 5-HT 1 DR mRNA and protein in the two brain regions (P<0.05).Manual acupuncture stimulation of GV 20, GB 20, etc. can inhibit the expression of RAMP 1 protein and mRNA in the STN and midbrain, and up-regulate the expression of 5-HT 1 DR in the two brain regions of migraine rats, which may be related to its effect in relieving migraine.