This experiment was conducted to evaluate the effects of different protease levels on performance, diarrhea rate, nutrient digestibility, blood metabolites, digestive enzyme activities, and intestinal health of weaned piglets. A total of 96 weaned piglets (Duroc × Landrace × Yorkshire, 28 d of age, initial average BW = 6.55 ± 0.17 kg) were divided into four groups (4 pigs/pen and 6 replicates/group) according to a completely random block design. Piglets were fed different levels of protease (0, 150, 300 or 600 mg/kg of diet) for 28 d. The results showed that the addition of protease had no significant effect on the growth performance of weaned piglets (p > 0.05), and the addition of 300 mg/kg protease significantly increased the apparent total intestinal digestibility (ATTD) of nutrients and the apparent ileal digestibility (AID) of amino acids in weaned piglets (p < 0.05), while the addition of 150 mg/kg and 600 mg/kg protease had no significant effect on the digestibility (p > 0.05). The nutrient digestibility of dry matter (DM), organic matter (OM), crude protein (CP) and total energy (GE) showed a trend of increasing and then decreasing with increasing protease concentration (p < 0.05). Adding 300 and 600 mg/kg protease significantly decreased serum ALB/GLO levels (p < 0.05) and duodenal pH (p < 0.05) and increased duodenal villus height (p < 0.05). The addition of protease significantly increased jejunal trypsin and chymotrypsin activities (p < 0.01) and duodenal and jejunal mucosal tight junction proteins in piglets. The mRNA expression levels of ZO-1 and CLAUDIN-1 in the duodenum together with ZO-1 and OCCLUDIN in the jejunum of piglets in the 300 and 600 mg/kg protease supplementation groups were significantly higher than those in the control group (p < 0.05). The results showed that, compared with the control group, protease could promote nutrient absorption, improve small intestine morphology and enhance digestive enzyme activity in weaned piglets. The suitable addition amount was 150–300 mg/kg in the present study.