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为研究大黄素(Em)对脂多糖(LPS)诱导的小鼠单核巨噬细胞系RAW264.7细胞向M1、M2型极化的影响。本研究设定RAW264.7细胞的LPS处理组、大黄素和LPS共同处理组和对照组,然后利用酶联免疫法检测各组细胞TNF-α、IL-12的分泌量;实时荧光定量PCR检测与巨噬细胞极化相关的精氨酸酶-1(Arg-1)、一氧化氮合酶(iNOS)、TNF-α、PPARγ的mRNA表达水平;Western-blot检测iNOS、TNF-α的蛋白表达。结果显示:LPS组与对照组比较其TNF-α、IL-12分泌增加,iNOS和TNF-α蛋白表达升高,表明细胞呈现M1型极化表型;大黄素能抑制LPS诱导的RAW264.7巨噬细胞分泌TNF-α和IL-12,抑制iNOS和TNF-α的mRNA及蛋白表达水平,具有促进IL-10、Arg-1、PPARγ基因mRNA水平升高的作用,使细胞表现为M2型极化表型。说明大黄素作为天然产物可能抑制LPS诱导的巨噬细胞向M1型极化作用,并促进M1表型巨噬细胞向M2表型极化,对于M1/M2亚型的失衡具有调节作用,有利于维持其动态平衡。
Abstract：To investigate the effect of emoxin(Em) on the polarization of RAW264.7 cell into M1-type and M2-type induced by lipopolysaccharide(LPS). In this study, it was divided into three groups: LPS treatment group, emodin and LPS co-treatment group and control group, and then the secretion of TNF-α and IL-12 in each group was detected by ELISA. Fluorescence quantitative PCR was used to detect the expression of arginase-1(Arg-1), nitric oxide synthase(INOS), TNF-α and PPARγ, which are related to macrophage polarization. The protein expression of iNOS and TNF-α was detected by Western-blot. The protein expression of nitric oxide synthase(iNOS) and TNF-α was detected by Western-blot. The results showed that compared with the control group, the secretion of TNF-α, IL-12 and expression of i NOS and TNF-α increased in the LPS group, indicating that the cells presented M1-type polarized phenotype.Emoesin can inhibit the secretion of TNF-α and IL-12 by LPS-induced RAW264.7 macrophages, and inhibit the mRNA and protein expression levels of i NOS and TNF-α, also promote the mRNA levels of IL-10, ARG-1 and PPARγgenes, and make the cells show the M2-type polarized phenotype. This suggests that emoxin, as a natural product, may inhibit the M1-type polarization of LPS-induced macrophages and promote macrophages to polarize from M1 phenotype to M2 phenotype, which has a regulatory effect on the imbalance of the M1/M2 subtype and is conducive to maintaining its dynamic balance.
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基本信息:
DOI：10.13417/j.gab.039.005304
中图分类号:R285.5
引用信息:
[1]包丽丽,张迪,宋利华,等.大黄素对LPS诱导小鼠巨噬细胞极化的影响[J].基因组学与应用生物学,2020,39(11):5304-5309.DOI:10.13417/j.gab.039.005304.
基金信息:
内蒙古自然科学基金面上项目(2015MS0888;2019MS08113);; 内蒙古自治区高等学校科学研究项目(NJZY17113;NJZZ20128);; 内蒙古自治区蒙医药协同创新培育中心项目基金(MYYXT201908);; 内蒙古医科大学博士启动基金(YKD2015BQ03)共同资助
2020-11-25
2020-11-25