Bombyx mori hemocyte extract has anti-inflammatory effects on human phorbol myristate acetate-differentiated THP-1 cells via TLR4-mediated suppression of the NF-κB signaling pathway

Abstract

Hemolymph is the circulating fluid of insects and is a key component of their immune system. However, little is known concerning hemocyte identification, development, differentiation and related cellular immune responses. The present study aimed to determine whether a hemocyte extract prepared from Bombyx mori larvae had anti-inflammatory effects; THP-1 (a human monocytic leukemia cell line) cells that had been differentiated into macrophage-like cells by treatment with phorbol myristate acetate (PMA) were used. THP-1 cells were cultured with different concentrations of a B. mori hemocyte extract prior to exposure to lipopolysaccharide (LPS) to induce an inflammatory response. The effects of the B. mori hemocyte extract on anti-inflammatory pathways were determined using reverse transcription-quantitative polymerase chain reaction and western blotting to assess the expression of pro-inflammatory molecules. The B. mori hemocyte extract inhibited the LPS-induced mRNA expression of Toll-like receptor 4 in addition to LPS-induced interleukin (IL)-1β, IL-6, IL-8 and tumor necrosis factor-α. Treatment of PMA-differentiated THP-1 cells with B. mori hemocyte extract also inhibited inducible nitric oxide synthase and cyclooxygenase-2 transcription and translation. Nuclear factor-κB activation and phosphorylation also decreased. Further in-depth functional studies are required to understand the mechanism underlying the anti-inflammatory effects of silkworm hemocyte extract.