Cervus nippon var. mantchuricus water extract treated with digestive enzymes (CE) modulates M1 macrophage polarization through TLR4/MAPK/NF-κB signaling pathways on murine macrophages

Abstract

The objective of this study was to investigate the effects of Cervus nippon var. mantchuricus water extract treated with digestive enzymes (CE) on the promotion of M1 macrophage polarization in murine macrophages. Macrophages polarize either to one phenotype after stimulation with LPS or IFN-γ or to an alternatively activated phenotype that is induced by IL-4 or IL-13. Cell viability of RAW264.7 cells was determined by WST-1 assay. NO production was measured by Griess assay. IL-6, IL-12, TNF-α, and iNOS mRNA levels were measured by RT-PCR. IL-6, IL-12, and IL-10 cytokine levels were determined by ELISA. TLR4/MAPK/NF-κB signaling in RAW264.7 cells was evaluated by western blotting. The level of NF-κB was determined by immunoblotting. CE induced the differentiation of M1 macrophages. CE promoted M1 macrophages to elevate NO production and cytokine levels. CE-stimulated M1 macrophages had enhanced IL-6, IL-12, and TNF-α. CE promoted M1 macrophages to activate TLR4/MAPK/NF-κB phosphorylation. M2 markers were downregulated, while M1 markers were upregulated in murine macrophages by CE. Consequently, CE has immunomodulatory activity and can be used to promote M1 macrophage polarization through the TLR4/MAPK/NF-κB signaling pathways.

PFS alleviates bone destruction in collagen-induced arthritis mice associated with macrophage polarization

“…This is supported by the inhibitory effect of PFS on the transcription factors NF-κB element p65 and activator protein-1 factor c-Fos, which are activated by the MAPK pathway. This finding may be further [48,49].…”

“…This is supported by the inhibitory effect of PFS on the transcription factors NF-κB element p65 and activator protein-1 factor c-Fos, which are activated by the MAPK pathway. This finding may be further validated in the mechanism of action of PFS on macrophage polarization and function in vitro experiments [48, 49].…”

Microenvironmental Analysis and Control for Local Cells under Confluent Conditions via a Capillary-Based Microfluidic Device

“…This device is also applicable for analysis of other secreted molecules, and Table S2 summarizes measurable molecules that are secreted from RAW264 cells. As the present device with ELISA detection allowed to detect and quantify TNF-α of 1 pg/mL (60 pM) or larger scale, the molecules in Table S2 are also measurable in their reported concentrations in the 0.1–100 nM scale. − Furthermore, this device produced adjacent stimulated and unstimulated cell groups, which simplified the comparison of the two groups by placing both groups in the same environment.…”