Objective To establish a skeletal muscle injury rat model using different amounts of eccentric exercise, and to observe the effects of the model on the expression of inflammatory genes in soleus and extensor digitorum longus muscles. Methods Male Wister rats were randomly divided into three groups: quiet control group (group A), one- time centrifugal exercise injury group (1Tgroup), and one-week centrifugal exercise injury group (1 W group). The rats used the animal treadmill to run downhill (centrifugal movement), and the gradient of the treadmill was - 16°. The 1 W group first received one week of adaptive training followed by one week of formal training. One week of adaptive training was given to the 1T group followed by one session of intensive centrifugal exercise. Twenty-four hours after the last training, all rats were fasted overnight for 12 hours, then weighed, anesthetized and sampled. Results Compared with group A, the serum creatine kinase and lactate dehydrogenase levels of rats in the 1T and 1 W groups were significantly higher (P< 0. 05 or P< 0. 01) and the serum CK and LDH levels of rats in the 1 W group were significantly lower than those in the 1T group (P< 0. 05 or P< 0. 01). There was no difference in mRNA levels of TLR4, MyD88, NF-κB, NLRP3, TNF-α, or IL-1β between soleus and extensor digitorum longus muscles in group A (P> 0. 05). The number of capillaries in the soleus was greater than that in the extensor digitorum longus in group A. The mRNA levels of TLR4, MyD88, NF-κB, NLRP3, TNF-α and IL-1β in the soleus of the 1T group were significantly higher than those in the soleus of group A (P< 0. 01). There was a small amount of inflammatory cell infiltration between the soleus muscle cells, and the intercellular space was increased. The mRNA levels of TLR4 and IL-1β in the extensor digitorum longus in the 1T group were significantly higher than those in group A ( P< 0. 01), and the mRNA levels of MyD88, NF-κB and NLRP3 in the extensor digitorum longus were significantly lower than those in the soleus in the 1T group (P < 0. 01). The mRNA levels of TLR4, MyD88 and NLRP3 in the soleus of the 1T group were significantly higher than those of group A (P< 0. 01), and the level of TLR4 mRNA was significantly lower than that of the 1T group (P< 0. 05). The level of TLR4 mRNA in the extensor digitorum longus in the 1 W group was significantly higher than that in group A (P< 0. 01), and the mRNA levels of MyD88 and NLRP3 were significantly lower than those in the soleus in the 1 W group (P< 0. 01). The microstructure of soleus and extensor digitorum longus muscles in the 1 W group was little changed. Conclusions One-time eccentric exercise and one week eccentric exercise can cause a certain degree of injury and inflammation in skeletal muscle of rats, which may be related to the activation of TLR4/ MyD88-related signaling stimulated by eccentric exercise. Compared with one week of centrifugation, one time high-intensity centrifugation caused more serious damage to skeletal muscle and higher expression of inflammatory factors. The degree of injury in the soleus of rats was greater than that in the extensor digitorum longus.