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目的探讨X盒结合蛋白1(X-box binding protein 1,XBP1)对脂多糖(lipopolysaccharide,LPS)处理的Kupffer细胞(Kupffer cells,KCs)TNF-α表达的影响。方法将BALB/c小鼠KCs随机区组法分为质粒组、阴性组和空白组。分别给予XBP1-shRNA、空白质粒转染和空白处理,并给予LPS。免疫细胞化学(SABC法)、Real-time PCR和Westernblot法检测各组KCs中XBP1基因及蛋白表达水平;酶联免疫吸附法(ELISA)检测核因子NF-κB活性及培养上清液中TNF-α含量。结果与阴性组、空白组相比,质粒组XBP1及下游因子TNF-α表达明显降低(P<0.05);而阴性组、空白组间比较比较无明显差异(P>0.05)。3组间NF-κB活性比较无明显差异(P>0.05)。结论 XBP1-shRNA质粒能明显抑制KCs XBP1基因的表达,并通过非NF-κB通路减少其下游因子TNF-α的释放。
Objective To investigate the effect of X-box binding protein 1 (XBP1) on the expression of TNF-α in Kupffer cells (KCs) treated with lipopolysaccharide (LPS). Methods The KCs of BALB / c mice were randomly divided into plasmid group, negative group and blank group. They were given XBP1-shRNA, blank plasmid transfection and blank treatment, and given LPS. The expression of XBP1 gene and protein in KCs of each group were detected by Real-time PCR and Western blotting using immunocytochemistry (SABC method). The activity of nuclear factor-kappaB (NF-κB) and the level of TNF-α in supernatant were detected by enzyme-linked immunosorbent assay (ELISA) α content. Results Compared with the negative group and the blank group, the expression of XBP1 and TNF-α in the downstream group was significantly decreased (P <0.05), but there was no significant difference between the negative group and the blank group (P> 0.05). NF-κB activity in the three groups showed no significant difference (P> 0.05). Conclusion XBP1-shRNA plasmid can significantly inhibit the expression of KBP XBP1 gene and decrease the release of its downstream factor TNF-α through a non-NF-κB pathway.