In Vitro Anti-Inflammatory Potential Of Tribolium Castaneum (Red Flour Beetle) Extract Using Human Red Blood Cell Membrane Stabilization And Heat-Induced Hemolysis Assays

Abstract

Inflammation is a fundamental biological response that protects the body against infection and tissue injury; however, persistent or excessive inflammation contributes to the development of numerous chronic disorders. The search for safer, naturally derived anti-inflammatory agents (plants and animals extract) has therefore become an important area of biomedical research. The present study investigated the in vitro anti-inflammatory activity of Tribolium castaneum (red flour beetle) extract using human red blood cell (HRBC) membrane stabilization and heat-induced hemolysis assays. A hydroethanolic extract of T. castaneum was prepared and evaluated at concentrations ranging from 10 to 50 µg/mL, with diclofenac sodium used as the reference anti-inflammatory drug. The extract exhibited concentration-dependent membrane-protective activity in both assays. In the HRBC membrane stabilization assay, the percentage inhibition of hemolysis increased from 7.85% at 10 µg/mL to 43.98% at 50 µg/mL, whereas diclofenac sodium achieved 65.45% inhibition at the highest concentration. Similarly, in the heat-induced hemolysis assay, the extract demonstrated inhibition ranging from 9.79% to 45.36%, compared with 65.46% for the standard drug. These findings indicate that T. castaneum extract possesses significant in vitro anti-inflammatory activity, likely through stabilization of erythrocyte membranes and protection against inflammatory damage. This study provides preliminary evidence supporting the therapeutic potential of insect-derived bioactive compounds as natural anti-inflammatory agents. Further studies are required to isolate the active constituents, elucidate their molecular mechanisms, and validate their efficacy and safety through in vivo and clinical investigations.