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.

中文翻译：

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乳铁蛋白抑制LPS诱导的RAW264.7细胞中HMGB1，microRNA 155、146和TLR4 / MyD88 /NF-кB通路的表达

摘要

目的

这项当前的研究评估了LF抵抗LPS激活的小鼠RAW264.7细胞中炎症性microRNA（miRNA），HMGB1表达和TLR4-MyD88-NF-кB通路的潜在机制。

方法

使用MTT测定法评估细胞代谢，并通过酶联免疫吸附测定法（ELISA）评估细胞因子（TNF-α，IL-6）的细胞培养水平。使用qPCR对miRNA的表达进行定量，分别通过Western blot和qPCR测定HMGB1，TLR4，MyD88和磷酸化的NF-κB（P-p65）的表达。

结果

结果表明LF下调IL-6和TNF-α表达。LF在LPS诱导的炎症反应中表现出P-p65的降解并减少了HMGB1，TLR4和MyD88的产生。重要的是，与抑制细胞因子和HMGB1-TLR4-MyD88-NF-кB途径同时，LF可能会诱导炎性选择的miRNA，mmu-mir -155和mmu-mir -146a表达减少。

结论

总而言之，这些发现提供了LF作为一种重要的抗炎药，可以调节HMGB1，mmu-mir -155，mmu-mir -146a和TLR4 / MyD88 /NF-кB途径。