摘要
目的调查锌指蛋白A20表达对人单核细胞LPS应答的影响,探讨A20可能的炎症调控作用与机制及其潜在的临床应用价值。方法采用细胞转染技术制备锌指蛋白A20人单核细胞转基因细胞模型和基因沉默细胞模型,以LPS攻击细胞,用适时荧光定量PCR技术测定锌指蛋白A20表达水平,用ELISA技术分析细胞核转录因子NF-κB活性及其依赖性炎性细胞因子TNF-α水平。结果锌指蛋白A20过表达组细胞遭受LPS攻击后其NF-κB活性和TNF-α水平明显低于LPS对照组(P<0.01),锌指蛋白A20基因表达抑制组细胞其NF-κB活性和TNF-α水平明显高于LPS对照组(P<0.01)。结论锌指蛋白A20对人单核细胞LPS应答具有明显的调节作用,可以明显降低炎症重要核转录因子活性和降低促炎细胞因子的表达。对脓毒症过度炎症反应实施干预具有潜在的临床应用价值。
Objective To investigate the effect of zinc （LPS） response in human monocytes so as to discuss its finger protein A20 expression on lipopolysaccharide regulation roles and mechanism to inflammation in vivo and its potential value of clinical application. Methods Cell transfection technique was respectively used to make zinc finger protein A20 transgenic cell model and gene silence cell model, both of which were then treated with LPS. Real-time reverse PCR was employed to analyze the expression of zinc finger protein A20 and ELISA to determine the activity of nuclear transcription factor NF-KB and the levels of dependent inflammatory cytokines including tumor necrosis factor （TNF） alpha. Results After LPS treatment, the NF-κB activity and the TNF-alpha level were significantly lower in the cells with overexpressed zinc finger protein A20 than LPS control cells （P 〈0.01 ）, but significantly higher in zinc finger protein A20 gene silenced cells （P 〈0. 01 ）. Conclusion Zinc finger protein A20 can effectively regulate LPS responses in human monocytes and markedly abate expressions of important nuclear factors and proinflammatory cytokines, proving that zinc finger protein A20 has potential clinical value for over inflammatory reaction of sepsis.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2007年第14期1388-1390,共3页
Journal of Third Military Medical University
基金
全军医学科研"十一五"计划面上项目(06MA189)~~
关键词
锌指蛋白A20
单核细胞
内毒素
RNA干扰技术
NF-ΚB
TNF-α
zinc finger protein A20
monocytes
lipopolysaccharide
RNA interference technique
nuclear factor-κB
tumor necrosis factor-α
