应用LiquiChip液相蛋白芯片技术检测HMGB1诱导肺泡巨噬细胞的细胞因子表达谱  

Expression spectrum of cytokines in HMGB1-stimulated alveolar macrophage detected by LiquiChip system

作　　者：丁宁[1] 肖慧[2] 许立新[1] 佘守章[1] 

机构地区：[1]广州市第一人民医院麻醉科,510180 [2]广州市第一人民医院门诊部,510180

出　　处：《临床麻醉学杂志》2010年第9期787-789,共3页Journal of Clinical Anesthesiology

基　　金：广东省医学科研基金项目(A2009497);广州市医药卫生科技一般引导项目(2009-YB-021);广州市医药卫生科技重点项目(2008-ZDi-14)

摘　　要：目的检测高迁移率族蛋白B1(HMGB1)诱导肺泡巨噬细胞(AM)的细胞因子表达谱。方法 RT-PCR扩增HMGB1 cDNA,插入pGEX4T-1原核表达载体并转大肠杆菌,IPTG诱导HMGB1表达,Ni2+-NTA和多粘菌素B亲和层析柱分离纯化。应用LiquiChip液相蛋白芯片检测HMGB1(20ng/ml)诱导AM细胞因子表达谱的变化。结果成功表达纯化的重组HMGB1可诱导AM肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、巨噬细胞炎性蛋白-1α(MIP-1α)、MIP-2、单核细胞趋化蛋白-1(MCP-1)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)及γ-干扰素(IFN)-γ的表达显著增加(P<0.01)。结论 HMGB1可诱导AM释放多种炎性细胞因子。Objective To detect the expression spectrum of cytokines in alveolar macrophages （AMs） induced by high mobility group box 1 （HMGB1） protein. Methods HMGB1 code gene was amplified by RT-PCR and cloned into vector pGEX4T-1 and transfected into E. coll. After induction by IPTG, the HMGB1 protein was purified with Ni2＋ -NTA chromatography and polymyxin B affinity column. The HMGB1 （20 ng/ml）-induced cytokine expression in AMs was detected using a LiquiChip system. Results The successfully purified recombinant HMGB1 induced high expression of TNF-α, IL-1β, IL-6, MIP-1α, MIP-2, MCP-1, GM-CSF and IFN-γ in AMs （P〈0.01）. Conclusion Inflammatory cytokines in AMs can be induced by HMGB1.

关 键 词：高迁移率族蛋白B1 肺泡巨噬细胞 细胞因子 

分 类 号：R563[医药卫生—呼吸系统]