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目的:观察TNF-α对脂多糖应答基因(lrg)在人HEK293和U937细胞中表达的影响.方法:正常培养人胚肾细胞(HEK293)和人单核细胞(U937),用TNF-α(终浓度1×106U/L)刺激2h.提取刺激前后HEK293和U937细胞的总蛋白,用纯化后的兔抗人Lrg抗血清作一抗(1∶1000),对TNF-α刺激前后的HEK293和U937细胞进行Western Blot分析.提取刺激前后HEK293和U937细胞的总RNA,用RT-PCR分析TNF-α对lrg在细胞中表达的影响.以β-actin为内参.结果:Western Blot分析显示,用TNF-α刺激2h后,lrg在人HEK293和U937细胞内的蛋白含量明显上升;RT-PCR结果显示,用TNF-α刺激2h后,人HEK293和U937细胞内的lrg mRNA水平明显上升.结论:TNF-α的刺激增强了人HEK293和U937细胞内lrg的表达,提示lrg可能参与了TNF-α诱导的炎症反应.
Objective: To observe the effect of TNF-α on the expression of LRP in human HEK293 and U937 cells.Methods: Normal human embryonic kidney cells (HEK293) and human monocytes (U937) were cultured with TNF-α The final concentration of 1 × 106U / L) stimulated for 2h.The total protein of HEK293 and U937 cells before and after stimulation were extracted and purified with purified anti-human Lrg antiserum (1: 1000), HEK293 U937 cells were analyzed by Western Blot.The total RNA was extracted from HEK293 and U937 cells before and after stimulation and RT-PCR was used to analyze the effect of TNF-α on the expression of lrg.Results: Western Blot analysis showed that using RT-PCR results showed that the level of lrg mRNA in human HEK293 and U937 cells increased significantly after TNF-α stimulation for 2h.Conclusion: The expression of lrg in human HEK293 and U937 cells induced by TNF- The stimulation of TNF-α enhanced the expression of lrg in human HEK293 and U937 cells, suggesting that lrg may be involved in the TNF-α-induced inflammatory response.