文献类型：期刊文献

中文题名：TLR-MyD88通路在COPD炎症反应中的作用及莱菔硫烷的抗感染效果

英文题名：The effect of Tlr-MyD88 pathway in inflammatory reaction of COPD and the anti-inflammatory effect of sulforaphane

作者：王婧[1,2];王敏[1,3];范玉敏[1,2];王燕[1,2]

第一作者：王婧

机构：[1]甘肃中医药大学继续教育学院,甘肃兰州730000;[2]酒泉职业技术学院医护系;[3]酒泉市人民医院呼吸内科

第一机构：甘肃中医药大学继续教育学院（职业技能培训中心）

年份：2018

卷号：38

期号：9

起止页码：2184

中文期刊名：中国老年学杂志

外文期刊名：Chinese Journal of Gerontology

收录：北大核心:【北大核心2017】；

基金：国家自然科学基金资助项目(No.81460709)

语种：中文

中文关键词：慢性阻塞性肺疾病;巨噬细胞;TLR4;MyD88;莱菔硫烷

外文关键词：Chronic obstructive pulmonary disease ; Macrophages; TLR4 ; MyD88 ; Sulforaphane

摘要：目的探讨TLR-MyD88通路在慢性阻塞性肺病(COPD)炎症反应中的作用及莱菔硫烷的抗感染效果。方法选取COPD稳定期患者64例为COPD组,另选同年龄段肺功能正常的体检者60名为非COPD组,分离两组外周血单核细胞,集落刺激因子诱导形成巨噬细胞,通过流式细胞术和荧光染色进行鉴定。COPD组巨噬细胞分为空白对照组、脂多糖组(终浓度为1 mg/L的脂多糖干预)、莱菔硫烷组(终浓度为30μmol/L的莱菔硫烷干预)、联合干预组。非COPD组巨噬细胞不作任何干预。实时荧光定量PCR和Western印迹法分别检测巨噬细胞中TLR4、MyD88 mRNA和蛋白表达;酶联免疫吸附实验(ELISA)检测细胞培养上清液中肿瘤坏死因子(TNF)-α、白细胞介素(IL)-6含量。结果荧光显微镜下观察细胞形态呈现巨噬细胞形态学特征,流式细胞术检测显示,外周血单核细胞中CD14表达量明显高于巨噬细胞(P<0.01)。基础状态下COPD患者巨噬细胞中TLR4、MyD88 mRNA和蛋白表达水平均明显高于非COPD组(P<0.05);COPD组巨噬细胞经脂多糖刺激后,TLR4、MyD88 mRNA和蛋白表达水平明显高于空白对照组(P<0.01);莱菔硫烷组、联合干预组均明显低于脂多糖组和空白对照组(P<0.01);莱菔硫烷组与非COPD组差异无统计学意义(P>0.05)。COPD患者TNF-α、IL-6浓度均明显高于非COPD组(P<0.05);脂多糖组均明显高于空白对照组(P<0.01);莱菔硫烷组、联合干预组均明显低于脂多糖组和空白对照组(P<0.01);莱菔硫烷组与非COPD组差异无统计学意义(P>0.05)。结论 COPD患者的巨噬细胞TLR-MyD88信号通路活化,下游炎性因子分泌量增加,提示该信号通路参与COPD炎症过程;莱菔硫烷可抑制TLR-MyD88信号通路,对COPD患者有抗感染作用。
Objective To investigate the effect of TLR-MyD88 pathway in inflammatory reaction of chronic obstructive pulmonary disease （COPD） and anti-inflammatory effect of sulforaphane. Methods The monocytes were selected from 64 stable COPD patients（ COPD group） and 60 healthy cases（ non COPD group）, the formation of macrophages were induced by colony-stimulating Which were identified by flow cytometry and fluorescence staining. The macrophages of COPD group was divided into control, LPS, sulforaphane and combination treatment group. The expressions of Tlr4, MyD88, mRNA and protein were detected by real-time quantitative PCR and Western blot, and the content of TNF-ct and IL-6 was detected by ELISA. Results The cell morphology showed morphological features of macrophage. The ex- pression of CD14 of macrophage was significantly higher than that in peripheral .blood mononuclear cells （P〈0.01）. At basic state, the Tlr4, MyDg8 mRNA and protein expression levels of macrophages of COPD were significantly higher than those in non COPD group （P〈 0.05） ; after stimulated by LPS, the Tlr4, MyD88 mRNA and protein expression level were significantly higher than those of control group （P〈0.01） ; the Tlr4, MyD88 mRNA and protein expression levels of sulforaphane and combination treatment groups were significantly lower than those of LPS and blank control groups （P〈0.01）. The concentrations of TNF-otand IL-6 in COPD group were significantly higher than those in non COPD group （P〈0.05） ; and they were significantly higher in LPS and control groups than those of sulforaphane and combination treatment groups （P〈0. 01 ）. Conclusions Tlr-MyD88 signal pathway is involved in the inflammatory process of COPD; sulforaphane could inhibit the Tlr-MyD88 signaling pathway, which has anti-inflammatory effect on patients with COPD.