TLR9信号通路参与急性肺损伤的作用机制研究  

Mechanism research on the role of TLR9 signaling pathways involved in acute lung injury

作　　者：冯健华[1] 

机构地区：[1]南方医科大学第五附属医院内四科,广州510900

出　　处：《国际医药卫生导报》2014年第11期1493-1495,共3页International Medicine and Health Guidance News

基　　金：广州市科技计划项目（201300000196）

摘　　要：目的研究Toll样受体-9在急性肺损伤中的作用机制，为临床诊疗提供理论依据。方法分离40例急性肺损伤患者的外周血单个核细胞，采用RT—PCR检测Toll样受体-9mRNA表达，外周血单个核细胞（PBMC）经过非甲基化胞嘧啶-鸟嘌呤二核苷酸序列的寡脱氧核苷酸（CpGODN）和不含非甲基化胞嘧啶-鸟嘌呤二核苷酸序列的寡脱氧核苷酸（non—CpGODN）、空白培养三种形式培养72h。采用3H—TdR掺入法测定PBMC增值变化，采用流式细胞仪检测T细胞亚群比例、T细胞表面CD69分子变化及CD8＋T细胞内IFN-γ和IL-4的表达。采用ELISA法对PBMC上清液IFN-α的浓度，并评价其对氯喹和抑制性ODN的反应程度。结果急性肺损伤患者PBMC表达TLR9mRNA的表达强度为（0．75±0．08），与健康组（0．76±0．09）比较差异无统计学意义（P〉0．05）；CpGODN诱导急性肺损伤患者PBMC增殖（P〈0．05），增加CD3＋T细胞表面CD69分子的表达（P〈0．05）；增强CD8＋T细胞产生IFN-γ的能力（P〈0．05）；增强CD4＋T／CD8＋T比值（P〈0．05）；CpGODN可促进IFN-α的分泌（P〈0．05）；氯喹和抑制性ODN会抑制CpGODN产生IFN—α。结论TLR9参与急性肺损伤患者的免疫调节，其激活效应包括促进PMBC活化、增加PBMC中CD4＋T的比例，诱导并增殖IFN-α产生，促进CD8＋T分泌IFN-γ。Objective To study the function mechanism of Toll -like receptor 9 in acute lung injury （ALI）, providing a theoretical basis for clinical diagnosis and treatment. Methods 40 cases of ALI with isolated peripheral blood mononuclear ceils, using RT-PCR detection of Toll-like receptor-9 mRNA expression in peripheral blood mononuclear cells （PBMC） after unmethylated cytosine-phosphate-guanine （CpG） sequence oligodeoxynucleotide （CpG ODN） and didn＇ t contain unmethylated CpG dinucleotide sequence oligodeoxynucleotide （non-CpG ODN）, three forms of blank culturing for72 h . Using 3H-TdR incorporation assay PBMC to get value changes, flow cytometry was used to detect the proportion of population using T ceils, T cells and CD8 surface expression of CD69 molecules change IFN- γ and IL-4＋T cells within. ELISA method was used to detect the IFN- α concentration in the supernatant of PBMC, and evaluate their degree of response to chloroquine and the inhibitory ODN. Results Patients with ALI PBMC expressed TLR9 mRNA expression level was （0.75 ± 0.08）, and the healthy group （0.76 ± 0.09） with no significant difference （P 〉 0.05）; CpG ODN induced PBMC proliferation in patients with ALI （P 〈 0.05 ） , increased expression of CD3＋T cell surface molecule CD69 （P 〈 0.05）; enhancing CD8＋T cells to produce IFN- γ in （P 〈 0.05）; enhanced D4＋T/CD8＋T ratio （P 〈 0.05 ）; CpG ODN promotes IFN- α secretion （P 〈 0.05）; suppressive ODN and chloroquine inhibit CpG ODN produced IFN-α. Conclusion TLR9 ALI patients involved in immune regulation, includes the promotion of its activation effect of PMBC, activating PBMC increase in the proportion of CD4＋T, and the proliferation induced by IFN- α production, promoting CD8＋T secreted IFN- γ.

关 键 词：TOLL样受体-9 CPG序列 急性肺损伤 

分 类 号：R563.8[医药卫生—呼吸系统]