Background. Delayed-onset muscle soreness (DOMS), also known as tenderness of touch, refers to the pain caused by muscle mechanical stimulation, such as contraction and stretching. Chinese massage has been widely used in the treatment of sports fatigue and sports injury, but there is controversy in the efficacy. In this experiment, we established DOMS model in rats to observe the prevention and treatment effect of massage, to find the best time for intervention, and to provide scientific basis for the prevention and treatment of exercise fatigue.Methods. 130 male SD(sprague-dawley, SD) healthy rats were randomly divided into blank group, control group and massage group. Except for blank group, the other rats received DOMS model. Professionals applied kneading and twisting methods on both lower limbs of rats. The expression of IL-2, IL-6 and IL-8 in skeletal muscle of rats were determined by western blot, PCR and ELISA, and the content of serum creatine kinase was determined by ELISA. In addition, we measured the concentration of Ca2+, Ca2+-ATPase in mitochondria of skeletal muscle. The changes of skeletal muscle structure were observed by scanning electron microscopy.Result. After massage, the expression of IL-2, IL-6, IL-8 and CK decreased compared with control group (P < 0.01), the expression of IL-2, IL-6 and IL-8 in post massage group was lower than that in front massage group (P < 0.01), and the content of CK in front massage group was lower than that in post massage group(P < 0.01). The content of Ca2+ in front massage 24, 48 and 72h group was lower than that in post massage (P < 0.01), the concentration of Ca2+-ATPase in front massage 24h and 72h group was lower than that in post massage group (P < 0.05).Conclusion. Massage can prevent the injury of muscle and reduce the inflammatory reaction of muscle after exercise. It can also improve the activity of Ca2+-ATPase, enhance the transport of Ca2+ by mitochondria and protect the skeletal muscle.