Objective To investigate the therapeutic effect of electroacupuncture at Guanyuan acupoint on neurogenic urinary retention through microRNA (miRNA)-199a-3p regulating receptor tyrosine kinase c-kit/extracellular signal-regulated kinase (ERK) pathway in rats. Methods Sixty model rats with neurogenic urinary retention were established by modified spinal cord transection method and divided into 5 groups by random number table method:model group, electroacupuncture group, miRNA-199a-3p antagomir (inhibitor) group, miRNA-199a-3p antagomir negative control group, and electroacupuncture+miRNA-199a-3p antagomir group, and another 12 SD rats were selected as sham group. After group intervention with drugs, the urodynamics of the rats were tested, and the maximum bladder volume, basal bladder pressure, and urine leakage point pressure were compared in each group; muscle strip experiment was used to detect the excitability of rat detrusor muscles and compare the spontaneous contraction frequency of muscle strips in each group; hematoxylin-eosin staining was used to detect the pathological and morphological changes of the bladder tissue of rats in each group; the kits were used to measure the serum levels of inflammatory cytokine cyclooxygenase 2 (COX-2), interleukin (IL-18) and inducible nitric oxide synthase (iNOS) of rats in each group; real-time fluorescence quantitative polymerase chain reaction experiment was used to detect the expression levels of miRNA-199a-3p and c-kit mRNA in the bladder tissue; Western blotting was used to detect the expression levels of c-kit/ERK pathway proteins in the bladder tissue. Results Compared with the sham group, the bladder tissue of the model group showed severe pathological damage, the maximum bladder volume, basal bladder pressure, urine leakage point pressure, serum COX-2, IL-18 and iNOS levels were significantly increased (all P<0.05), the spontaneous contraction frequency of muscle strips, the expression levels of miRNA-199a-3p and c-kit mRNA in bladder tissue, and the expression levels of phosphorylated (p)-ERK/ERK and c-kit in bladder tissue were significantly decreased (all P<0.05). Compared with the model group and the electroacupuncture+miRNA-199a-3p antagomir group, the pathological damage of the bladder tissue of rats in the electroacupuncture group was reduced, the maximum bladder volume, basal bladder pressure, urine leakage point pressure, serum COX-2, IL-18 and iNOS levels were decreased (all P<0.05), the spontaneous contraction frequency of muscle strips, the expression levels of miRNA-199a-3p and c-kit mRNA in bladder tissue, and the expression levels of p-ERK/ERK and c-kit in bladder tissue were increased (all P<0.05); the pathological damage in the miRNA-199a-3p antagomir group was aggravated, the maximum bladder volume, basal bladder pressure, urine leakage point pressure, serum COX-2, IL-18 and iNOS levels were increased (all P<0.05), the spontaneous contraction frequency of muscle strips, the expression levels of miRNA-199a-3p and c-kit mRNA, and the expression levels of p-ERK/ERK and c-kit were decreased (all P<0.05). Compared with the model group, there was no significant difference in the indicators in the miRNA-199a-3p antagomir negative control group (all P>0.05). Conclusion Electroacupuncture at Guanyuan acupoint can activate c-kit/ERK pathway by up-regulating the expression of miRNA-199a-3p, thereby reduce the inflammatory damage of bladder tissue in neurogenic urinary retention rats, enhance the excitability of detrusor, repair the bladder function, and improve the symptoms of urinary retention in rats.