Detection of TNF-α mRNA Expression in Lipopolysaccharide-stimulated NR8383 Cells Using Real-Time Fluorescence Quantitative PCR

Objective To detect the expression of TNF-α mRNA in lipopolysaccharide(LPS)-induced NR8383 alveolar macrophages using real-time fluorescence quantitative PCR.Methods Alveolar macrophages were treated with 1 μg·mL-1LPS(LPS group) or phosphate-buffered saline(control group).Total RNA was extracted with TRIzol reagent and TNF-α mRNA expression was examined by real-time quantitative PCR using SYBR Green Ⅰ.Results Nonspecific amplification was not detected.Compared with control group,the expression of TNF-α mRNA in LPS-stimulated group increased 124.1 times(P0.01).Conclusion Real-time fluorescence quantitative PCR is a highly specific and reliable method for the detection of TNF-α mRNA.