Department of Genetic, Islamic Azad University - Tabriz Branch, Tabriz, Iran.
Stem Cells Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Immunopharmacol Immunotoxicol. 2021 Apr;43(2):153-159. doi: 10.1080/08923973.2021.1872616. Epub 2021 Jan 12.
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.
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.
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, -155, and -146a expression.
Altogether, these findings provide LF as a prominent anti-inflammatory agent that could modulate HMGB1, -155, -146a, and TLR4/MyD88/NF-кB pathway.
本研究旨在评估 LF 对 LPS 激活的 RAW264.7 细胞中炎症 microRNAs（miRNAs）、HMGB1 表达和 TLR4-MyD88-NF-кB 通路的潜在机制。
采用 MTT 法评估细胞代谢，采用酶联免疫吸附试验（ELISA）检测细胞培养上清液中细胞因子（TNF-α、IL-6）的水平。采用 qPCR 定量检测 miRNAs 的表达，采用 Western blot 和 qPCR 分别检测 HMGB1、TLR4、MyD88 和磷酸化 NF-кB（P-p65）的表达。
结果表明 LF 下调了 IL-6 和 TNF-α 的表达。LF 可降解 P-p65，并减少 LPS 诱导的炎症反应中 HMGB1、TLR4 和 MyD88 的产生。重要的是，与抑制细胞因子和 HMGB1-TLR4-MyD88-NF-кB 通路的同时，LF 还可诱导炎症相关 miRNA（-155 和 -146a）表达下调。
综上所述，这些发现表明 LF 可作为一种有潜力的抗炎药物，通过调控 HMGB1、-155、-146a 和 TLR4/MyD88/NF-кB 通路发挥作用。
