Background : Acetylcholine is related with various functions, including cognition and behavior, and increased activity of cholinesterase has been reported in the brains of people suffering from Alzheimer’s disease (AD). As such, the inhibition of cholinesterase activity could be a means of ameliorating neuronal degenerative diseases such as AD. Reactive oxygen species (ROS) can cause neuronal cell damage. The AchE inhibitory effects of Sorghum bicolor (SB) have been revealed by research. This study was conducted to compare the cholinesterase inhibitory effects and anti-oxidative effects of SB extracts according to their extraction conditions. Methods and results : Eight extracts were prepared from SB seed, which was extracted using three different methods including room temperature extraction, reflux extraction at 85℃, and accelerate solvent extraction (ASE) at 50℃ by using distilled water and/or ethanol as a solvent. AchE and BuchE inhibition activities of the extracts were measured in vitro, and their inhibitory activities on ROS, nitric oxide (NO) production and cell proliferation were analyzed in lipoppolysacchride–treated BV2 mouse microglia cells. According to the results of the experiments, the 50% ethanol extract obtained by room temperature extraction showed a BuchE inhibitory effect of 40% at the final concentration of 100 ㎍/㎖, while the other 50% ethanol extracts showed a BuchE inhibitory effect of around 20%. The 100% Ethanol extract obtained from reflux extraction at 50 ㎍/㎖ showed the highest inhibitory effect on NO generation as 58.3%, whereas the 50% ethanol extract obtained from ASE extraction at 50 ㎍/ ㎖ showed the highest inhibitory effect on ROS generation as 56.0%. Conclusion : The results of the experiments show that the 50% and 100% ethanol extracts prepared under different temperature, pressure and solvent conditions have more effective on strong cholinesterase inhibition, anti-oxidative and anti-inflammatory effects..