BACKGROUND: Pediatric tuina is an effective alternative therapy for managing spleen deficiency diarrhea. The application of spleen-meridian and large-intestine-meridian points has been shown to alleviate symptoms and has garnered some support from evidence-based medicine. Nonetheless, there remains a dearth of research elucidating the biological effects of these specific acupoints. This study aimed to explore their effects by focusing on the material basis involving mast cells (MCs). METHODS: The experimental design induced spleen-deficiency diarrhea in Kunming mice through a 7-day administration of RESULTS: The treatment combining spleen-meridian and large-intestine-meridian points markedly ameliorated diarrhea symptoms and improved fecal scores in Kunming mice exhibiting spleen deficiency. Pediatric tuina treatment facilitated the restoration of the colonic barrier and reduction in MC counts within the skin acupoint-brain-gut axis, consequently affecting the biologically active substance tryptase. CONCLUSION: This study reveals the biological mechanism underlying the efficacy of specific acupoints in pediatric tuina, employing a holistic perspective encompassing the skin-brain-gut axis and MCs. Our findings substantiate the scientific basis for the effectiveness of tuina therapy in managing diarrhea and offer a new avenue for fundamental research on specific acupoints within pediatric tuina.