【摘要】 目的:探讨小干扰RN(A siRNA)对小鼠脾淋巴细胞分泌的肿瘤坏死因子α(TNF-α)的抑制效果。方法:体外PCR扩增合成针对TNF-α靶位点的DNA表达盒,转染经脂多糖(LPS)激活的小鼠脾淋巴细胞,在细胞内源性RNA聚合酶III作用下转录形成siRNA,诱发目标mRNA的降解。于转染后48 h收集细胞培养上清。TNF-α浓度用ELISA方法测定。结果:与空白对照组和无关干扰组相比,干扰组TNF-α分泌量明显降低(P<0.05),抑制率约为16%。结论:RNA干扰技术可以在原代培养的小鼠脾淋巴细胞中发挥特异性干扰作用,产生抑制TNF-α分泌的效果,为TNF-α的基因治疗开拓新途径。更多还原

【Abstract】 Objective: To investigate the inhibitory effect of small interfering RNA(siRNA) targeting tumor necrosis factor-α(TNF-α) gene in mice spleen lymphocytes.Methods: DNA expression cassette targeting TNF-α was synthesized by polymerase chain reaction in vitro,transfected into lymphocytes activated by lipopolysaccharide(LPS),and transcribed by endogenous RNA polymerase III to form siRNA and then evoke targeted mRNA degradation.The cell culture supernatant was collected 48 hours after the transfection,and the TNF-α concentration in the supernatant was detected by enzyme-linked immunosorbent assay.Results: Compared with blank and positive control groups,the expression of TNF-α in interference group significantly decreased(P < 0.05),and the inhibitory rate was about 16%.Conclusion: RNA interference technology can produce specific interference effect in mice spleen lymphocytes of primary culture and inhibit the secretion of TNF-α.更多还原