X盒结合蛋白1对脂多糖处理的Kupffer细胞炎性因子表达的影响  

Influence of X-box binding protein 1 on expression of inflammatory factor in lipopolysacharide-induced Kupffer cells

作　　者：李铁军[1] 刘作金[2] 李生伟[2] 刘长安[2] 屈谦 薛强[2] 龚建平[2] 

机构地区：[1]重庆市第九人民医院普外科,重庆400700 [2]重庆医科大学附属第二医院肝胆外科,重庆400010 [3]重庆市卫生局,重庆401147

出　　处：《第三军医大学学报》2011年第14期1455-1457,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金(30772098)~~

摘　　要：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 explore the influence of X-box binding protein 1（XBP1） on the expression of inflammatory factor of lipopolysaccharide（LPS）-induced Kupffer cells（KCs）.Methods KCs derived from BALB/c mice were randomly divided into an XBP1-shRNA group,a negative control group and a blank control group,which received XBP1-shRNA plasmid transfection,control vector transfection and no treatment,respectively.Then the three groups were all treated with LPS.The mRNA and protein expression of XBP1 in KCs was determined by Real-time PCR,Western blotting and immunohistochemistry.The activity of NF-κB and the density of TNF-α in the supernatant of coculture were assessed by enzyme-linked immunosorbent assay（ELISA）.Results The expression of XBP1 and its downstream factor TNF-α was significantly lower than that in the negative control group and blank control group（P0.05）.However,there was no significant difference between the two control groups（P0.05）.In addition,the activity of NF-κB in the three groups had no significant difference（P0.05）.Conclusion XBP1-shRNA plasmid can significantly inhibit the expression of XBP1 gene and reduce the release of its downstream factor TNF-α without changing the activity of NF-κB in KCs.

关 键 词：X盒结合蛋白1 KUPFFER细胞 肿瘤坏死因子-Α 小鼠 

分 类 号：R322.47[医药卫生—人体解剖和组织胚胎学] R341[医药卫生—基础医学]