Department of Nutrition and Food Science, University of Maryland, 0112 Skinner Building, College Park, MD 20742, USA.
Inflamm Res. 2012 Dec;61(12):1329-38. doi: 10.1007/s00011-012-0533-8. Epub 2012 Jul 28.
In vitro cell culture models are widely used in inflammation research; however, information regarding the time- and dose-dependency of inflammatory responses toward LPS in these cell lines is scattered in the literature.
J774A.1 mouse macrophage and THP-1 human monocyte cell lines.
J774A.1 and THP-1 cells were treated with 0-500 ng/mL lipopolysaccharide for 0-24 h.
SRB and BCA tests were used to measure total protein. Real-time PCR was used to determine gene expression levels, and ELISA was used to assess the protein levels. One-way ANOVA and Tukey's Honestly Significant Difference test were used to test the significance levels.
In J774A.1 and THP-1 cells, cytokines responded in distinct patterns upon LPS stimulation in a time- and dose-dependent manner, and the differential regulation of the response to LPS between J774A.1 and THP-1 cells appeared to correlate with the differential regulation of TLR4 at the mRNA level.
In summary, this study indicated that temporal and dose-dependent responses to LPS need to be controlled for and that extrapolation of data on mechanisms may differ between cell lines of different origin.
体外细胞培养模型广泛应用于炎症研究；然而，关于这些细胞系中 LPS 诱导炎症反应的时间和剂量依赖性的信息在文献中较为分散。
J774A.1 小鼠巨噬细胞和 THP-1 人单核细胞系。
用 0-500ng/ml 脂多糖处理 J774A.1 和 THP-1 细胞 0-24 小时。
SRB 和 BCA 试验用于测量总蛋白。实时 PCR 用于测定基因表达水平，ELISA 用于评估蛋白水平。采用单因素方差分析和 Tukey 的Honestly Significant Difference 检验来测试显著性水平。
在 J774A.1 和 THP-1 细胞中，细胞因子对 LPS 刺激的反应呈时间和剂量依赖性的独特模式，J774A.1 和 THP-1 细胞对 LPS 反应的差异调节似乎与 TLR4 在 mRNA 水平的差异调节相关。
综上所述，本研究表明，需要控制 LPS 的时间和剂量依赖性反应，并且不同来源的细胞系之间的机制数据推断可能存在差异。
