Background: To explore the immediate effects and mechanism of acupuncture on craving after tobacco cessation based on the fractional amplitude of low-frequency fluctuation (fALFF). Methods: This was a functional magnetic resonance imaging (fMRI) study. Forty participants were recruited and divided into a smoking group and a non-smoking group, 20 cases in each one. The smoking participants were requested to quit smoking 24 hours before the EMRI scan. The scan process was scan - acupuncture - scan. Between the two scans, acupuncture was applied at Lieque (LU 7), Hegu (LI 4), Baihui (GV 20), Zusanli (ST 36), Sanyinjiao (SP 6) and Taichong (LR 3) in two groups. Results: After acupuncture, self-made craving for smoking, Minnesota Nicotine Withdrawal Scale (MNWS) and Questionnaire of Smoking Urges (QSU) were all reduced (P<0.05). The fMRI results indicated the immediate effects of acupuncture on smoking craving were significant, and identified salience network (SN) consisted of anterior cingulate cortex and insula, prefrontal cortex, visual cortex and cerebellum as key brains area. Correlation analysis indicated that NWNS scores were positively correlated with the mean fALFF in the ACC (P<0.05) and negatively correlated with the mean fALFF in the insula (P<0.05) in the smoking group. Conclusions: This was the first study in China to explore the neural mechanisms of acupuncture for smoking craving. The results indicated that the effects of acupuncture on smoking craving were significant, and the SN played a critical role in the process.