Screening and Activity Comparison of Anti-inflammatory Components of Walnut Proteins
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Abstract:
The fractions of walnut proteins with the highest anti-inflammatory activity and the effect of gastrointestinal digestion on their properties and anti-inflammatory activity were studied. Six walnut protein protease hydrolysates were investigated for their anti-inflammatory activity using the lipopolysaccharide (LPS)-induced RAW 264.7 cell model. The protease hydrolysate demonstrating the strongest anti-inflammatory activity underwent fractionation using ultrafiltration technology, and the anti-inflammatory potential of fractions with different molecular weights was investigated. An in vitro simulated gastrointestinal digestion model was established to evaluate the effects on the degree of hydrolysis, peptide content, and anti-inflammatory activity of the most active fraction. Results indicated that all six protease hydrolysates inhibited nitric oxide (NO) release from LPS-induced RAW 264.7 cells. Notably, the alkaline protease hydrolysate exhibited the highest inhibitory effect, achieving an inhibition rate of 18.33% at a dose of 800 μg/mL. Within the alkaline protease hydrolysate, the fraction with molecular weight<1 ku demonstrated the most effective inhibition of NO release, with an inhibition rate of 25.25% at the same dose. Comparative analysis revealed that the peptide content of the<1 ku-simulated gastrointestinal digestion group increased from 5.56 g/100 g to 10.93 g/100 g. Likewise, the degree of hydrolysis increased from 1.82% to 7.54%, with an increase in the NO release inhibition rate from 24.75% to 46.50%. These results underscore the highest anti-inflammatory activity observed in the alkaline protease hydrolysate of walnuts with molecular weight<1 ku, which was further enhanced post-gastrointestinal digestion. This study provides foundational insights for the isolation and purification of anti-inflammatory peptides from walnuts, and offers a theoretical basis for the valorization of walnut by-products.
