Objective: To observe the clinical efficacy of row needling at the Governor Vessel on the head for poststroke insomnia, analyze the changes of sleep structure, serum amino acid neurotransmitters, vascular endothelial cell function and cognitive function, in order to explore the mechanism of this therapy.Methods: A total of 41 patients with poststroke insomnia were included in an observation group and treated with row needing at the Governor Vessel on the head; 33 patients with poststroke insomnia were included in a control group and given routine acupuncture treatment. The patients in both groups were treated consecutively for one month. The efficacy was observed and the changes of related indicators were compared. Results: After treatment, the efficacy of the observation group was better than that of the control group (P<0.05). The sleep latency in the observation group was shorter than that in the control group (P<0.05); the total sleep duration, deep sleep duration, light sleep duration and rapid eye movement sleep duration in the observation group were significantly longer than those in the control group (P<0.05); the total waking duration and the waking times over 5 min were significantly less than those in the control group (P<0.05). The serum glutamic acid, γ-aminobutyric acid, calcitonin gene-related peptide, brain-derived neurotrophic factor, nerve growth factor and nitric oxide levels were significantly higher than those in the control group (P<0.01). The tumor necrosis factor-α, interleukin-6, bradykinin, and endothelin-1 levels were lower than those in the control group (P<0.01). The Pittsburgh sleep quality index score in the observation group was lower than that in the control group (P<0.01), and the mini-mental state examination score was higher than that in the control group (P<0.05). Conclusion: Row needling at the Governor Vessel on the head can effectively improve the sleep structure, cognitive function, and sleep quality of patients with poststroke insomnia. The possible mechanisms are regulating serum amino acid neurotransmitters and improving the function of vascular endothelial cells.