Antioxidant Activities of <i>Tenebrio molitor</i> Hydrolysates and Their Effects on the Expressions of Pro-inflammatory Cytokines in the Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

Tenebrio molitor (TM), an edible insect rich in bioactive proteins, has been reported to possess antioxidant and anti-inflammatory properties.This study hypothesized that enzymatic hydrolysis of TM using complementary proteases would enhance these biological activities.Accordingly, TM proteins were hydrolyzed using Alcalase , Protamex , or a combination of both enzymes.The resulting hydrolysates were designated as TM-A, TM-P, and TM-AP, respectively and assessed for their degree of hydrolysis (DH), antioxidant capacity, and effects on cell viability of RAW 264.7 macrophages.The combined treatment with both enzymes significantly increased DH, Brix values, and overall antioxidant activity compared to singleenzyme treatments or untreated controls.All TM hydrolysates enhanced cell viability of RAW 264.7 macrophages, with TM-AP exhibiting the most pronounced effect on lipopolysaccharidestimulated macrophages.Furthermore, TM-A and TM-AP selectively suppressed interleukin 1 beta expression without significantly affecting that of interleukin 6 or tumor necrosis factor alpha.These findings suggest that dual-enzyme hydrolysis augments the antioxidant potential and promotes the expressions of anti-inflammatory cytokines, supporting their potential as functional food ingredients for inflammation-related conditions.