Objective: This study aimed to realize the high-value utilization of biological active ingredients and anti-inflammatory peptides from the Pinctada martensii meat. Methods: The hydrolysis process of the P. martensii meat was optimized by single factor test and response surface test with degree of hydrolysis (DH) as the index. The amino acid composition and anti-inflammatory activity of P. martensii meat hydrolysate peptides were evaluated and analyzed. Results: The results showed that neutral protease was the optimal enzyme, and the optimal hydrolysis conditions were solid-liquid ratio 1∶1 (g/mL), temperature 46.3 ℃, hydrolysis time 1.4 h, and substrate ratio 0.3%. The hydrolysis degree was 22.88%, which was not significantly different from the theoretical value, and the regression model was reliable. The hydrolysates including 19.84% of essential amino acids, 21.19% of hydrophobic amino acids and 10.46% of positively charged amino acids. In LPS induced RAW264.7 anti-inflammatory model of mouse macrophages, the hydrolyzed peptide of P. martensii had no cytotoxicity and was beneficial to the proliferation of macrophages in the mass concentration range of 0~4.0 mg/mL. At the mass concentration of 2.0 mg/mL, hydrolyzed peptide could effectively inhibit the production of NO and inflammatory cytokines TNF-α, IL-6 and IL-1β; the inhibition rate of NO, TNF-α, IL-6 and IL-1β was 70.00%, 83.01%, 85.04% and 83.11%, respectively. Conclusion: Hydrolytic peptide of P. martensii has a complete range of amino acids, which can effectively inhibit the production of NO and inflammatory cytokines TNF-α, IL-6 and IL-1β in RAW264.7 macrophages, showing good anti-inflammatory activity.