| 摘要: |
| ［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。 |
| 关键词: RNA干扰 siRNA表达框架 转录后基因沉默 肿瘤坏死因子α |
| DOI：10.11724/jdmu.2008.05.04 |
| 分类号:Q78 |
| 基金项目: |
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| Effect of RNA interference to excretion of TNF-α in activated peritoneal macrophage of mice |
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SUN Jie1, GAO Ying2, JIANG Miao-na1, ZHENG Ning-ning3, SONG Xiao-yu3, ZHANG Hai-peng3
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1.Department of Pathophysiology,Dalian Medical University,Dalian 116044, China;2.Department of Pharmacy,Shenyang Medical College,
Shengyang 110034,China;3.Department of Pathophysiology,China Medical University,Shengyang 110001,China
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| Abstract: |
| ［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. |
| Key words: RNA interference siRNA expression cassettes post-transcriptional gene silencing tumor necrosis factor alpha |
