摘要
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。
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-αand 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-αand PPARγ,which are related to macrophage polarization.The protein expression of iNOS and TNF-αwas detected by Western-blot.The protein expression of nitric oxide synthase(iNOS)and TNF-αwas 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-αincreased in the LPS group,indicating that the cells presented M1-type polarized phenotype.Emoesin can inhibit the secretion of TNF-αand 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.
作者
包丽丽
张迪
宋利华
纳仁高娃
王志钢
Bao Lili;Zhang Di;Song Lihua;Naren Gaowa;Wang Zhigang(School of Life Sciences,Inner Mongolia University,Hohhot,010010;Department of Basis Medical,Inner Mongolia Medical University,Hohhot,010059)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2020年第11期5304-5309,共6页
Genomics and Applied Biology
基金
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。
作者简介
通信作者:纳仁高娃,narensan128@hotmail.com;通信作者:王志钢,lswzg@imu.edu.cn。
