OBJECTIVE: To observe and analyze the changes in brain amplitude of low-frequency fluctuation (ALFF) in resting-state induced by ST36 (Zusanli) point acupuncturing (Ac), and to discuss its central modulating mechanism. METHODS: Fifteen healthy young volunteers received ST36 Ac and non-acupoints Ac respectively, the resting-state fMRI data were acquired before and 20 min after the withdrawal of AC. ALFF of encephalic regions were calculated using REST software, and one-sample t-test analysis was performed on the intragroup data acquired after Ac by statistical parametric mapping 5 (SPM5), to get the maps of ALFF changes induced by ST36 Ac and non-acupoints Ac respectively for analyzing the difference between them finally. RESULTS: ST36 Ac increased ALFF in widespread brain areas, including cerebral cortex (frontal, temporal, parietal and occipital lobe), brainstem, cerebellum, etc., with the amplitude and extent higher than those increased by non-acupoint Ac, showing statistical significance in ALEF of right cerebellar posterior lobe, fusiform gyrus, central posterior gyrus, central anterior gyrus, supramarginal gyrus, inferior parietal lobule, middle frontal gyrus (P < 0.05). Meanwhile, ALFF in brain areas of left temporal pole, superior temporal gyrus, mid-temporal gyrus, uncus, bilateral forepart of callosal gyrus, right amygdala, right inferior gyri orbitales were decreased by Ac, also showing significant difference between ST36 Ac and non-acupoint Ac (P < 0.05). CONCLUSION: ST36 Ac could induce ALFF changes in multiple functional regions of brain,and its mechanism is probably produced by modulating the respective functional center and complex network in brain to influence the target organs.