【摘要】： 目的:研究靶向IL-1β基因的RNA干擾對巨噬細胞分泌炎癥因子IL-1β、TNF-α、IL-6等的影響。 方法:設(shè)計并合成3條靶向小鼠IL-1β基因的siRNA。脂質(zhì)體法轉(zhuǎn)染小鼠巨噬細胞株RAW264.7,并設(shè)立對照組。采用Real-time PCR和ELISA方法檢測其對經(jīng)脂多糖刺激的RAW264.7細胞表達IL-1β、TNF-α、IL-6等炎癥因子的影響,篩選基因沉默效率最高的干擾序列。 結(jié)果:在mRNA水平和蛋白水平,siRNA1、siRNA2可有效抑制經(jīng)脂多糖刺激的RAW264.7細胞表達IL-1β。但其對TNF-α、IL-6 mRNA水平表達無顯著差異。 結(jié)論:脂質(zhì)體介導(dǎo)的靶向小鼠IL-1β基因RNA干擾可有效沉默小鼠巨噬細胞IL-1β基因表達。
[Abstract]:Aim: to study the effects of RNA interference targeting IL-1 尾 gene on the secretion of inflammatory factors IL-1 尾, TNF- 偽 and IL-6 by macrophages. Methods: three siRNA. targeting mouse IL-1 尾 genes were designed and synthesized. The mouse macrophage cell line RAW264.7, was transfected with lipofectamine and the control group was established. The effects of Real-time PCR and ELISA on the expression of IL-1 尾, TNF- 偽, IL-6 and other inflammatory factors in RAW264.7 cells stimulated by lipopolysaccharide (LPS) were detected, and the interference sequences with the highest gene silencing efficiency were screened. Results: at the level of mRNA and protein, siRNA1,siRNA2 could effectively inhibit the expression of IL-1 尾 in RAW264.7 cells stimulated by lipopolysaccharide. However, there was no significant difference in the expression of TNF- 偽 and IL-6 mRNA. Conclusion: targeting mouse IL-1 尾 gene RNA interference mediated by liposome can effectively silence the expression of IL-1 尾 gene in mouse macrophages.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R392

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