Abstract: Objective: To analyze the anti-inflammatory effect on TNF-α induced MH7A cells before and after deer sinew enzymolysis. Methods: Proliferative inhibitory activity of each peptide segment from deer sinew natural protein (DSNP) and deer sinew enzymolysis protein (DSEP) in ultrafiltration fraction was determined by MTT assay, and the components with best activity were screened. The effects of active component on the secretion of inflammatory factors (NO and IL-6) by MH7A cells were determined by ELLSA, and analysised the amino acid composition and content before and after deer sinew enzymolysis. Multivariate statistical analysis was used to find out the relationship between the differential amino acids before and after deer sinew enzymolysis and the inhibition of MH7A cells secreting inflammatory factors. Results: DSNP-5 and DSEP-5 had the best inhibitory effect on the proliferation of MH7A cells (P<0.05), and inhibited the secretion of NO and IL-6. When the concentration was 25~200 μg/mL, the release of inflammatory factors (NO and IL-6) in DSNP-5 was lower than that in DSEP-5, with significant difference (P<0.05), indicating that DSNP-5 had better inhibition ability.The amino acid composition before and after deer sinew enzymolysis was basically the same, but the content was significantly different. Gly was not only a characteristic difference component, but also closely related to the inhibition of MH7A cells secreting inflammatory factors NO and IL-6. The Gly content of DSNP-5 (31.01%) was higher than that of DSEP-5 (28.91%). Conclusion: Gly is the difference amino acid related to anti-inflammatory activity before and after deer sinew enzymolysis. The effect of enzymolysis on anti-inflammatory activity of deer sinew is explored, which provide reliable basis for the development and application.