IL-17对小鼠脊髓星形胶质细胞相关炎性因子及A20、NF-κB蛋白表达的影响  

A20 and NF- κB: a possible regulator in IL- 17 stimulated astrocyte inflammatory cytokines

作　　者：李柯柯[1] 宗少晖[1] 曾高峰[2] 方晔[1] 

机构地区：[1]广西医科大学第一附属医院脊柱骨病外科,广西南宁530021 [2]广西医科大学公共卫生学院,广西南宁530021

出　　处：《中国矫形外科杂志》2016年第16期1486-1491,共6页Orthopedic Journal of China

基　　金：国家自然科学基金资助项目(编号:81560351)

摘　　要：[目的]探讨白介素17(Interleukin-17,IL-17)对小鼠脊髓星形胶质细胞相关炎性因子产生及A20(tumor necrosis factorαinducible protein 3,TNFAIP3)、NF-κB蛋白表达的影响。[方法]使用不同浓度的IL-17刺激体外培养小鼠脊髓星形胶质细胞,并于不同时间点(3、6、12、24和48 h)行酶联免疫吸附检测(ELISA)法和qRT-PCR检测IL-6、TNF-α、MIP-2、MCP-1/5的表达,采用蛋白质免疫印迹法(WB)和qRT-PCR检测A20、NF-k B的表达。[结果]与对照组相比,实验组小鼠脊髓星形胶质细胞在IL-17刺激6、12 h时,IL-6、TNF-α、MCP-1/5和MIP-2 mRNA和蛋白的表达水平分别显著升高(P<0.05);NF-κB mRNA和蛋白的表达分别在6 h和12 h时显著升高,A20蛋白在12 h时表达量显著减少与NF-κB呈相反趋势,但其mRNA的表达呈逐渐减少趋势。[结论]A20和NF-κB可能在小鼠脊髓星形胶质细胞相关炎性因子产生过程中起调节作用。[Objective]To investigate the effect of IL- 17 on the expressions of astrocyte- related inflammatory cytokines,A20 and NF- κB regulatory proteins in a mice model. [Method]Three- day new born mice was sacrificed to isolate the spinal cord. Different concentrations of IL- 17 were used to stimulate the cultured astrocytes. Then,the expression of IL- 6,TNF- α,MCP- 1 /5 and MIP- 2 were determined using qRT- PCR and ELISA kits at different time points（ 3 h,6 h,12 h,24 h and 48 h after surgery）. Western blotting（ WB） and RT- PCR were carried out to measure the expression of A20（ tumor necrosis factor α inducible protein 3,TNFAIP3） and NF- κB. [Result]Compared with the result from the control group,the mRNA and protein expression levels of IL- 6,TNF- α,MCP- 1 /5 and MIP- 2 increased significantly at 6h and 12 h respectively（ p〈0.05）. The mRNA expression level of NF- κB increased and peaked at 6 h before gradually declining,while the expression ofA20 decreased. The protein expression level of NF- κB increased and peaked at 12 h,while the expression of A20 had an opposite response. [Conclusion]A20 and NF- κB may have participating roles on regulating the expression and secretion of inflammatory mediators.

关 键 词：白介素17 星形胶质细胞 炎性因子 A20 NF-ΚB 

分 类 号：R651.2[医药卫生—外科学]