摘要
[目的]利用siRNA表达框架研究RNA干扰技术对小鼠腹腔活化巨噬细胞分泌TNF-α和IL-1的抑制作用及其干扰序列的特异性。[方法]原代培养小鼠腹腔巨噬细胞,构建针对小鼠TNF-α和IL-1的siRNA表达框架,转染细胞,用ELISA方法检测TNF-α和IL-1分泌情况。[结果]TNF-α干扰组TNF-α分泌量(21.87±1.20)pg/mL较IL-1干扰组(28.02±1.03)pg/mL及对照组(27.64±1.92)pg/mL均明显降低,差异有显著性意义(P<0.05)。IL-1干扰组IL-1分泌量(40.37±4.02)pg/mL较TNF-α干扰组(57.86±3.91)pg/mL及对照组(59.55±3.57)pg/mL均明显降低,差异有显著性意义(P<0.05)。[结论]构建的针对TNF-α和IL-1的siRNA表达框架能够明显抑制巨噬细胞分泌TNF-α和IL-1,且干扰作用具有序列特异性。
[ Objective] To study the specifical effect of RNA interference to inhibit the excretion of TNF- α and IL- 1 in mice activated peritoneal macrophages with siRNA expression cassettes. [ Methods ] Stimulate the primary culture of peritoneal macrophages with LPS. Prepare siRNA expression cassettes that target TNF - α and IL - 1, then transfect them to macrophages. Collect the cell culture upper liquid to assay the concentration of TNF - α and IL - 1 α with ELISA. [ Resuits ] The concentration of TNF - α of cell culture upper liquid in TNF - α interference group（ 21.87 ± 1.20 ） pg/mL is ob- viously lower than that in IL - 1 interference group（ 28.02 ± 1.03 ） pg/mL （ P 〈 0. 05 ） and that in negative - control group （27.64 ± 1.92） pg/mL （ P 〈 0. 05 ）. The concentration of IL - 1 of cell culture upper liquid in IL - 1 interference group （40.37 ± 4.02 ） pg/mL is obviously lower than that in TNF - α interference group（ 57.86 ± 3.91 ） pg/mL （ P 〈 0. 05 ） and that in negative - control group（ 59.55 ± 3.57 ） pg/mL （ P 〈 0.05 ）. [ Conclusion ] siRNA expression cassettes that aim TNF - α and IL - 1 target genes produce specific interference effect in inhibitory excretion of TNF - α and IL - 1 in mice macrophages.
出处
《大连医科大学学报》
CAS
2008年第6期527-529,共3页
Journal of Dalian Medical University
关键词
RNA干扰
siRNA表达框架
肿瘤坏死因子Α
白细胞介素-1
RNA interference
siRNA expression cassettes
post -transcriptional gene silencing
tumor necrosis factor al- pha
interleukin - 1
