RNA干扰对小鼠腹腔活化巨噬细胞分泌TNF-α的影响

Effect of RNA interference to excretion of TNF-α in activated peritoneal macrophage of mice

[目的]探讨RNA干扰对小鼠腹腔活化巨噬细胞表达TNF-α的抑制作用。[方法]原代培养小鼠腹腔巨噬细胞,构建针对小鼠TNF-α的siRNA表达框架,转染细胞,用RT-PCR方法检测TNF-αmRNA的水平,用ELISA方法检测TNF-α分泌情况。[结果]从凝胶电泳图像TNF-α/β-actin灰度值比可以看出干扰组TNF-αmRNA的水平0.18±0.03较阳性对照组0.91±0.05及阴性对照组0.85±0.10明显降低(P<0.05)。干扰组TNF-α分泌量(21.87±1.20)pg/mL较阳性对照组(28.02±1.03)pg/mL及阴性对照组(27.64±1.92)pg/mL均明显降低(P<0.05)。[结论]RNA干扰可以有效地抑制原代培养小鼠腹腔巨噬细胞TNF-αmRNA水平及其蛋白分泌。

[Objective] To investigate the inhibitory effect of RNA interference to the excretion of TNF-α in activated peritoneal it macrophages of mice.[Methods] Culture peritoneal macrophages with LPS.Prepare siRNA expression cassettes of RNA interference group and then transfect it macrophages.Assay the mRNA of TNF-α,with collect the cell culture upper liquid to assay the concentration of TNF-α with ELISA.[Results] RT-PCR the amount of TNF-α mRNA in interference group 0.18±0.03 is obviously lower than that in positive-control group 0.91±0.05（P〈0.05） and in negative-control group 0.85±0.10（P〈0.05）.The concentration of TNF-α of cell culture upper liquid in interference group（21.87±1.20）pg/mL is obviously lower than that in positive-control group（28.02±1.03） pg/mL（P〈0.05） and in negative-control group（27.64±1.92）pg/mL（P〈0.05）.[Conclusion] RNAi can interfere the mouse peritoneal macrophage to express TNF-α mRNA.