Objective To explore the analgesic initiation mechanism of three-manipulation and three-acupoint tuina in model rats with minor chronic constriction injury (CCI). Methods Fifty-six SD rats were divided randomly into eight groups: normal group, sham group, model 1 group, model 2 group, tuina 1 group, tuina 2 group, tuina 1 + transient receptor potential vanilloid-1 (TRPV1) antagonist group, and tuina 2 + transient receptor potential ankyrin 1 (TRPA1) antagonist group. The model, tuina, and tuina + antagonist groups were established with minor CCI models. The tuina and tuina + antagonist groups received the three-method three-point intervention ( point method, dial method, kneading method, Yinmen point, Chengshan point, Yanglingquan point) 7 days after modeling. The model and sham groups were subjected to grasping restraint, and the normal group received no intervention. After the respective interventions, each group was tested for changes in mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) to detect different types of pain. The nitric oxide (NO) content of the dorsal root ganglion (DRG) was determined by the nitrate reductase method, and changes in protein and gene expression levels of components of the TRPV1 / TRPA1-NO-cGMPprotein kinase G (PKG) signaling pathway in the DRG of each group were determined by enzyme-linked immunosorbent assay, Western blot, and qPCR. Results Compared with the model group, MWT and TWL were prolonged in the tuina 1 and tuina 2 groups. Expression levels of TRPV1, TRPA1, NO, soluble guanylate cyclase-β, cGMP, and PKG1 in the DRG were significantly decreased in the tuina 1, tuina 2, tuina 1 + TRPV1 antagonist, and tuina 2 + TRPA1 antagonist groups. Conclusions Tuina can effectively improve the symptoms of thermal and mechanical hyperalgesia caused by peripheral nerve injury after one-time intervention. Tuina can exert immediate and continuous analgesic effects via the TRPV1 / TRPA1-NO-cGMP-PKG signaling pathway.