Lactoferrin suppresses LPS-induced expression of HMGB1, microRNA 155, 146, and TLR4/MyD88/NF-кB pathway in RAW264.7 cells

Abstract

Objective

This current study evaluated the underlying mechanisms of LF against the inflammatory microRNAs (miRNAs), HMGB1 expression, and TLR4-MyD88-NF-кB pathway in LPS-activated murine RAW264.7 cells.

Methods

MTT assay was used to assess cell metabolism and the cell culture levels of the cytokines (TNF-α, IL-6) were evaluated by Enzyme-linked immunosorbent assay (ELISA). The expression of miRNAs was quantified by using qPCR and the expression of HMGB1, TLR4, MyD88, and phosphorylated NF-κB (P-p65) were determined with Western blot and qPCR, respectively.

Results

The results indicated that LF downregulates IL-6 and TNF-α expression. LF exhibited the degradation of P-p65 and reduced the production of HMGB1, TLR4, and MyD88 in LPS-induced inflammatory response. Importantly, in parallel with the suppression of cytokines and HMGB1-TLR4-MyD88-NF-кB pathway, LF could induce a decrease in inflammatory selected miRNAs, mmu-mir-155, and mmu-mir-146a expression.

Conclusions

Altogether, these findings provide LF as a prominent anti-inflammatory agent that could modulate HMGB1, mmu-mir-155, mmu-mir-146a, and TLR4/MyD88/NF-кB pathway.

Keywords:

• Lactoferrin
• TLR4-MyD88-NF-кB
• HMGB1
• MicroRNA
• RAW264.7

Author contributions

Yousef Faridvand and Mohammad Nouri conceived and designed the research; Maryam Nemati, Saeideh Akseh, Maryam Amiri, Hamid Reza Nejabati, Ahmadreza Jodati, Nazila Fathi Maroufi, and Yousef Faridvand performed the experiments; Yousef Faridvand analyzed the data; Hamid Reza Nejabati drafted the text and wrote the paper.

Disclosure statement

No potential conflict of interest was reported by the author(s).