In order to investigate the analgesic effect of aqueous extract of male rhinoceros beetle on mice, male rhinoceros beetle powder was treated with aqueous extract, and ICR mice were randomly divided into blank control group, positive drug aspirin group, aqueous extract of male rhinoceros beetle with low, medium and high concentrations. The effects of acetic acid on the number of acetic acid torsions and the pain quotient of mice were investigated by the acetic acid-induced writhing, tail-flicking and hot plate methods. The results showed that the writhing times of mice in low, medium and high concentration groups were（11.17 ± 5.77）,（15.00 ± 6.00）,（11.83 ± 4.57）respectively, and the percentage of analgesic in the low concentration group reached 51.43%. Comparing with the model control group, the low, medium and high concentration groups of male rhinoceros beetle aqueous extract could significantly reduce the effect of acetic acid on the number of mouse writhing times, and enhance the mice's pain threshold; prolonged the heat resistance time of mice in hot plate and tail-flick experiments.This experiment results proved that the aqueous extract of male rhinoceros beetle did have certain analgesic effects on mice without toxic side effects, which provided some theoretical basis for the development of rhinoceros beetle as medicine and new analgesic drugs in the future.