脂多糖对人单个核细胞白细胞介素-6基因表达的影响  

Effects of lipopolysaccharides on the expression of IL-6 gene in human peripheral blood mononuclear cells

作　　者：邹立林[1] 吕建新[1] 

机构地区：[1]温州医学院检验医学与公共卫生学院细胞与分子医学研究所,浙江温州325027

出　　处：《温州医学院学报》2004年第1期31-33,共3页Journal of Wenzhou Medical College

摘　　要：目的 :探讨脂多糖 (LPS)对人单个核细胞表达白细胞介素 6 (IL 6 )mRNA的影响。方法 :抽取人外周静脉血 ,经EDTA抗凝 ,LPS刺激培养 ,分离单个核细胞并提取总RNA ,将总RNA逆转录为cDNA ,然后用IL 6引物进行热启动PCR扩增。扩增产物热变性后与IL 6检测探针、捕获探针进行夹心杂交 ,最后加入亲和素 辣根过氧化物酶及其底物 ,显色检测杂交信号。IL 6引物及探针用PrimerPremier 5 .0软件设计。结果 :LPS可在转录水平上诱导IL 6mRNA表达 ,其作用强度与LPS剂量呈正相关 ;IL 6mRNA表达丰度有时间效应 ,刺激 4h后 ,IL 6mRNA的表达最高。结论 :LPS能显著增强人单个核细胞表达IL 6mRNA。Objective:To investigate the effects of lipopolysaccharides(LPS) on the expression of IL-6 mRNA in human peripheral blood mononuclear cells. Methods:Human peripheral blood mononuclear cells (PBMC) were isolated from whole blood treated with ethylenediaminetetraacetic acid (EDTA) and stimulated by LPS. Total RNA extracted from the PBMC was reversely transcripted to cDNA,which used as templates for hot-start PCR amplification of IL-6. Sandwich hybridization occurs between IL-6 amplicon,capture probe and detection probe. The hybridization signals were detected utilizing avidin-horseradish peroxidase and its substracts. Specific IL-6 primers and probes were designed using Primer Premier 5.0 software.Results:LPS is an effective inducer which can increase IL-6 mRNA. Expression of IL-6 mRNA is positively correlated with the concentration of LPS. Abundance of IL-6 mRNA varied with the time of LPS*0*8s stimulation and peaked at 4 h.Conclusion:LPS can enhance the expression of IL-6 mRNA in human PBMC.

关 键 词：脂多糖 逆转录-聚合酶链反应 酶联夹心杂交 白细胞介素-6 核糖核酸 信使 

分 类 号：Q786[生物学—分子生物学]