Abstract： Objective： To explore the mechanism of moxibustion in alleviating cancer-related fatigue （CRF）. Methods：Forty SPF male BALB/c mice were randomly divided into five groups （n=eight per group）：the blank control group， the model group， the 38 ℃ moxibustion group， the 44 ℃ moxibustion group， and the 44 ℃moxibustion + inhibitor group. A CRF model was established by inducing CT-26 subcutaneous transplanted tumors followed by intraperitoneal chemotherapy with 5-fluorouracil （5-FU）. After successful modeling， moxibustion intervention was applied at Guanyuan （CV4） point， with the local skin temperature controlled at （38±1） ℃ or （44±1） ℃ . The intervention was performed once daily for 10 minutes，five days a week with a two-day break，for a total of two weeks. The tail suspension immobility time and the maximum grip force of forelimbs of mice were tested after successful tumor inoculation， after chemotherapy completion， and after two weeks of intervention. Organ indices were calculated. The mRNA expression of transient receptor potential vanilloid 1 （TRPV1） protein in the skin of Guanyuan point，and the mRNA levels of transcription factor T-box expressed in T cells （T-bet） and GATA binding protein 3 （GATA-3） in the spleen were detected by qPCR. Serum levels of interleukin （IL）-2， interferon-γ （IFN-γ）， IL-4， and IL-6 were detected by enzyme-linked immunosorbent assay （ELISA）. Results：Body weight in the blank control group increased over time. In all other groups，body weight initially increased，then decreased after chemotherapy，and subsequently increased after intervention. Compared with the model group， 38 ℃ moxibustion group， and 44 ℃ moxibustion + inhibitor group，tumor weight was reduced in the 44 ℃ moxibustion group （P＜0.05）. Compared with the blank control group， the model group showed prolonged tail suspension immobility time and decreased maximum grip force of forelimbs after chemotherapy completion and after two weeks of intervention （P＜0.05）. Compared with the model group， 38 ℃ moxibustion group，and 44 ℃ moxibustion + inhibitor group，the 44 ℃ moxibustion group showed shortened tail suspension immobility time and increased maximum grip force of forelimbs after two weeks of intervention （P＜0.05）. Compared with the blank control group，the model group exhibited increased spleen index，GATA-3 mRNA，IL-4， and IL-6 （P＜0.05），and decreased thymus index，T-bet mRNA，T-bet/GATA-3 ratio，IL-2，and IFN-γ （P＜ 0.05）. Compared with the model group，38 ℃ moxibustion group，and 44 ℃ moxibustion + inhibitor group，the 44 ℃ moxibustion group showed increased thymus index，TRPV1 mRNA，T-bet mRNA，T-bet/GATA-3 ratio，IL-2，and IFN-γ （P＜0.05），and decreased GATA-3 mRNA，IL-4，and IL-6 （P＜0.05）. Conclusion：Moxibustion had the effect of relieving CRF in mice with colon cancer，especially moxibustion at 44 ℃. Its mechanism may involve activating the TRPV1 target，regulating immunity via the Ca2+/CaN/NFAT signaling pathway，restoring the T helper （Th）1/Th2 balance，thereby alleviating CRF.