The purpose of this study was to investigate the effects of 8 weeks of resistance training (RT) on activity of total and A₁₂ Type of acetyl cholinesterase in Soleus muscles of rats. 16 male wistar rats provided from Razi institute (age: 10 weeks and weight: 172.415±7.090 gr), were randomly divided to 2 groups (Control; n=8 and RT; n=8). Training group carried out 8 weeks (5 session/week) of resistance training on 1-meter height ladder (divided by 26 stairs) with loading 30% of their body weight (suspended from the tail) in the first week which was gradually increased to 200% in the last week. Training included 3 sets of 4 reps with 3 min rest between sets. 48 hrs after last session of training, Sol muscles of animals moved out under sterilized situation by cutting on posterio-lateral side of hind limb. For separating AchE subunits, we used from homogenization and electrophoresis (0/06 non-denaturaing Polyacrilamide). Acetyl cholinesterase activity was measured by Elisa kit. Independent t-test showed that there was no significant differences between training and control groups in both total (p=0.262) and A₁₂ forms (p=0.246) of AchE in soleus muscle. The reason for insignificant differences in acetyl cholinesterase of soleus might be indicative of no complete involvement of this muscle in this type of training and therefore no responsiveness of acetyl cholinesterase activity of this muscle following resistance training. However, this should be studied in future with higher volume and intensity of training. Because the increases in AchE content by training is not excluded yet.