Amelioration of Toll-like Receptor-4 Signaling and Promotion of Mitochondrial Function by Mature Silkworm Extracts in Ex Vivo and in Vitro Macrophages

Nutrients. 2024 Nov 18;16(22):3932. doi: 10.3390/nu16223932.

Abstract

Objectives: The unknown immune-enhancing effects of steamed mature silkworms (Bombyx mori L.), known as HongJam (HJ), were investigated.

Methods: Supercritical fluid extracts from the White Jade variety of HJ (WJ-SCEs) were applied to in vitro RAW264.7 macrophages (RAWMs) and ex vivo bone marrow-derived macrophages (BMDMs).

Results: WJ-SCE enhanced the proliferation and viability of both RAWMs and BMDMs. Supplementation with WJ-SCE significantly reduced the lipopolysaccharide (LPS)-induced expression of iNOS mRNA and protein, resulting in decreased nitric oxide (NO) production. Additionally, WJ-SCE lowered the mRNA and protein expression of COX-2 and reduced the levels of pro-inflammatory cytokines. The mitochondrial function, ATP levels, and reactive oxygen species levels in LPS-treated macrophages were restored following WJ-SCE treatment. WJ-SCE modulated LPS-Toll-like receptor-4 (TLR-4) signaling by reducing the levels of phosphorylated (p)-p38, p-ERK1/2, and p-p65. WJ-SCE also restored gene expression related to cytokines, chemokines, glucose transport, mitochondrial metabolism, and TLR-4 signaling, suggesting the inhibition of pro-inflammatory M1 macrophage polarization. Furthermore, WJ-SCE enhanced macrophage phagocytic and pinocytotic activity.

Conclusions: WJ-SCE demonstrated anti-inflammatory effects by inhibiting LPS-induced M1 polarization in both macrophage types, potentially suppressing chronic inflammation while enhancing phagocytosis and pinocytosis.

Keywords: HongJam; chronic inflammation; phagocytosis; pinocytosis; supercritical fluid extracts.

MeSH terms

  • Animals
  • Bombyx*
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cytokines / metabolism
  • Lipopolysaccharides
  • Macrophages* / drug effects
  • Macrophages* / metabolism
  • Mice
  • Mitochondria* / drug effects
  • Mitochondria* / metabolism
  • Nitric Oxide / metabolism
  • Nitric Oxide Synthase Type II / metabolism
  • RAW 264.7 Cells
  • Reactive Oxygen Species / metabolism
  • Signal Transduction* / drug effects
  • Toll-Like Receptor 4* / metabolism

Substances

  • Toll-Like Receptor 4
  • Tlr4 protein, mouse
  • Nitric Oxide
  • Lipopolysaccharides
  • Cytokines
  • Reactive Oxygen Species
  • Nitric Oxide Synthase Type II