早期生长反应基因1对巨噬细胞分泌肿瘤坏死因子α的影响  

Influence of early growth response gene-1 on tumor necrosis factor alpha in macrophages

作　　者：牛海艳[1,2] 王金胜[2] 马志健[3] 黎岳南[1] 

机构地区：[1]海南医学院病理学教研室,海南省海口市571101 [2]中南大学湘雅医学院病理学系,湖南省长沙市410078 [3]海南医学院解剖学教研室,海南省海口市571101

出　　处：《中国组织工程研究与临床康复》2008年第50期9875-9878,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：海南省自然科学基金指导性项目(80438)~~

摘　　要：背景:体内外研究已证实肺巨噬细胞合成和分泌肿瘤坏死因子α等参与肺组织局部损伤和炎症反应,但具体发生机制仍不明确。目的:观察核转录因子早期生长反应基因1对巨噬细胞分泌肿瘤坏死因子α的影响。设计、时间及地点:随机分组设计,于2004-06/2006-12在湘雅医学院病理学系完成。材料:标准石英粉尘(SiO2)为Sigma公司产品;早期生长反应基因1抗体为Santa Cruz公司产品;小鼠巨噬细胞系RAW264.7购自中科院上海生物细胞研究所细胞库。方法:实验将巨噬细胞分为正常对照组、二氧化硅刺激组、Egr-1+二氧化硅组和IgG+二氧化硅组,前两组加无血清培养基,后两组分别加入5,10,20mg/L不同浓度的Egr-1和IgG抗体干预。主要观察指标:酶联免疫吸附实验检测各组细胞上清中肿瘤坏死因子α蛋白的水平;反转录-聚合酶链反应检测细胞内肿瘤坏死因子αmRNA的表达。结果:①与二氧化硅刺激组相比,不同浓度Egr-1+二氧化硅组细胞上清中肿瘤坏死因子α蛋白水平均下降(P<0.01),且10mg/L组与5,20mg/L组相比,差异有显著性意义(P<0.05)。不同浓度IgG+二氧化硅组相比,上清液中肿瘤坏死因子α蛋白水平差异无显著性意义(F=1.008,P=0.438)。②二氧化硅刺激组肿瘤坏死因子αmRNA表达高于正常对照组(P<0.01),而Egr-1+二氧化硅组mRNA表达低于二氧化硅刺激组和IgG+二氧化硅组(P<0.01)。结论:二氧化硅致巨噬细胞分泌肿瘤坏死因子α增加可能通过激活核转录因子早期生长反应基因1介导的信号通路而实现。BACKGROUND： The activation of early growth response gene-1 （Egr-1） and the production of tumor necrosis factor-α （TNF- α ） in macrophages participate local lung tissue injury and inflammation. However, the mechanism of action is still uncertain. OBJECTIVE： To explore the effect of signai pathway mediated by Egr-1 on the TNF-α secretion in macrophages. DESIGN, TIME AND SETTING： Randomized design. The experiment was performed at Department of Pathology, Xiangya Medical School from June 2004 to December 2006. MATERIALS： SiO2 and the antibody against Egr-1 were products of Sigma and Santa Cruz biotechnology, Inc. Cell line RAW264.7 came from the Cell Bank in Shanghai. METHODS： Macrophages were divided into normal control, SiO2, Egr-1＋ SiO2 （5,. 10 and 20 mg/L antibody against Egr-1） and IgG＋ SiO2 groups （5, 10 and 20 mg/L IgG）. MAIN OUTCOME MEASURES： The expression of TNF- α protein in cell supernatant was measured by enzyme-linked immunosorbent assay （ELISA）. The expression of TNF-α mRNA was monitored by reverse transcriptase-polymerase chain reaction （RT-PCR）. RESULTS： Compared with SiO2 group, the expression of TNF-α protein in cell supernatant stimulated by Egr-1 and silica was reduced （P 〈 0.01）, and there were significant differences between 10 mg/L group and 5 and 20 mg/L groups （P 〈 0.05）. No differences were found in the expression of TNF-α protein among IgG＋ SiO2 groups （F=1.008, P=0.438）. TNF-α mRNA expression in SiO2 group was significantly higher than in normal control group （P 〈 0.01）. In addition, TNF-ct mRNA expression in Egr-1＋ SiO2 groups was significantly lower than SiO2 group and IgG＋ SiO2 groups （P 〈 0.01）. CONCLUSION： The expressions of proteins and mRNAs of TNF-α are up-regulated by SiO2 through activating the signal pathway mediated by Egr- 1.

关 键 词：二氧化硅 巨噬细胞 早期生长反应基因1 肿瘤坏死因子Α 

分 类 号：R563[医药卫生—呼吸系统]