siRNA表达框架对TNF-α和IL-1的特异性抑制

siRNA expression cassettes specifically inhibit excretion of TNF-α and IL-1 in mice macrophages

关键词:  RNA干扰  siRNA表达框架  肿瘤坏死因子α  白细胞介素-1

Key words:  RNA interference  siRNA expression cassettes  post-transcriptional gene silencing  tumor necrosis factor alpha  interleukin-1

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。

Abstract:

［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-1α with ELISA.［Results］The concentration of TNF-α of cell culture upper liquid in TNF-α interference group(21.87±1.20)pg/mL is obviously 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 ).［Conclusions］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.