GSK-3β对小鼠骨髓树突状细胞成熟和功能的调控作用  

Role of glycogen synthase kinase 3β in maturation and function of murine myeloid dendritic cells in vitro

作　　者：褚帅[1] 李海侠[1] 李欣[1] 康霞[1] 黄清水[1,2] 王红霞[1] 裘宇容[1] 

机构地区：[1]南方医科大学南方医院检验医学科,广东广州510515 [2]南昌大学第一附属医院检验科,江西南昌330006

出　　处：《南方医科大学学报》2015年第12期1809-1814,共6页Journal of Southern Medical University

基　　金：广东省高等学校科技创新项目(2012KJCX0029)

摘　　要：目的探讨GSK-3β对小鼠骨髓树突状细胞(BMDC)成熟和功能的调控作用。方法脂多糖催熟BMDC,Western blotting检测刺激前后糖原合成酶激酶-3β(GSK-3β)的磷酸化水平的改变;利用GSK-3β的选择性抑制剂SB216763处理BMDC,检测表型、细胞因子表达和混合淋巴反应(MLR)的变化情况;构建过表达小鼠GSK-3β的慢病毒载体并转染DC2.4细胞,Western blotting检测过表达GSK-3β对调控树突状细胞(DC)成熟的关键调控因子鸟的网状内皮组织增生病毒癌基因相关B(Rel B)蛋白水平的影响。结果经脂多糖处理后,GSK-3βY216磷酸化水平下调,Ser9磷酸化水平显著上调,表明GSK-3β的活性显著下降。抑制GSK-3β的活性能上调DC表面共刺激分子CD40和CD86的表达,削弱脂多糖诱导的促炎细胞因子IL-6和IL-12表达上调,而对抗炎细胞因子IL-10的表达有促进作用,同时降低脂多糖诱导成熟的DC刺激同种异基因T细胞增殖的能力。在未成熟的DC2.4细胞中,过表达GSK-3β能够下调Rel B的水平。结论 GSK-3β参与调控DC的成熟和功能,高活性的GSK-3β抑制未成熟树突状细胞(i DC)的自发性成熟,GSK-3β失活后促进DC表型的成熟,而在脂多糖诱导DC分化的过程中GSK-3β发挥促炎作用。GSK-3β能够下调Rel B的蛋白水平,有望成为构建新型耐受性DC的新靶点。Objective To investigate the role of glycogen synthase kinase 3β（GSK-3β） in the maturation and function of murine bone marrow- derived dendritic cells（BMDCs）. Methods Mature DCs（m DCs） induced by LPS were examined for GSK- 3βphosphorylation level with Western blotting before and after LPS exposure. To explore the role of GSK-3β in maturation and function of DCs, we added SB216763, a selective inhibitor of GSK-3β, in the cell culture of immature DCs（i DCs）, and examined CD40 and CD86 expressions in the cells by flow cytometry and the expression of IL- 6, IL- 12 and IL- 10 m RNA by real- time PCR; the changes of the immunogenicity of the cells was evaluated by mixed lymphocyte reaction. The expression of GSK-3βand Rel B was examined by Western blotting in DC2.4 cells transfected with a lentiviral vector over-expressing murine GSK-3βgene. Results LPS exposure significantly lowered GSK-3β activity in i DCs as demonstrated by increased Ser9 phosphorylation and reduced Tyr216 phosphorylation. GSK- 3β inhibition induced DC maturation by increasing the expression of surface costimulatory molecules CD40 and CD86, lowered the expressions of IL-6 and IL-12 while enhanced the expression of IL-10 in i DCs, and impaired mixed lymphocyte reaction of the cells. In DC2.4 cells, lentivirus- mediated over- expression of GSK- 3βobviously down-regulated the expression of Rel B. Conclusion GSK-3β is a crucial enzyme involved in the differentiation and maintenance of an immature phenotype of DCs. GSK- 3β is constitutively active in i DCs to inhibit their spontaneous maturation. DCs become phenotypically mature after inhibition of GSK-3β, which also executes a proinflammatory task in DC activation. The reduction of Rel B protein levels as a result of GSK- 3β overexpression supports GSK- 3β as a new target for inducing tolerogenic DCs.

关 键 词：糖原合成酶激酶-3Β 树突状细胞 成熟和功能 鸟的网状内皮组织增生病毒癌基因相关B 

分 类 号：R392[医药卫生—免疫学]