低氧对巨噬细胞分泌TNF-α和IL-6的影响及其机制  

EFFECTS OF HYPOXIA ON THE SECRETIONS OF PROINFLAMMATORY CYTOKINES TNF-αAND IL-6 IN MACROPHAGES AND ITS MECHANISM

作　　者：张翠萍[1] 谢印芝[1] 陈鹏[2] 洪欣[1] 聂鸿靖[1] 董宏彬[1] 尹昭云[1] 

机构地区：[1]军事医学科学院卫生学环境医学研究所,天津300050 [2]军事医学科学院放射医学研究所,北京100850

出　　处：《中国应用生理学杂志》2005年第3期281-284,共4页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金(30393130)

摘　　要：目的:观察低氧对巨噬细胞(M)前炎症因子TNF-α和IL-6分泌的影响及其机制。方法:收集分离小鼠腹腔M,建立M的低氧(1%O2,5%CO2)培养模型,并用非特异性酯酶染色法进行鉴定;ELISA法检测上清液中TNF-α和IL-6的含量;RT-PCR法检测TNF-α和IL-6的转录物水平;用Westernblot法检测M核内NF-κB的激活量;通过在培养液中加入氢化可的松(5mg/L),观察低氧时TNF-α和IL-6分泌量的变化。结果:TNF-α和IL-6分泌量在低氧12h时明显增加(P<0.01);低氧6h时,TNF-αmRNA和IL-6mRNA表达量明显高于对照组(P<0.01);M核内NF-κB的激活量在低氧2h时明显增高(P<0.05),低氧5h内持续存在;而当培养液中加入氢化可的松抑制NF-κB活性后,TNF-α和IL-6的分泌水平无明显变化。结论:低氧可通过核转录因子NF-κB途径促进细胞因子TNF-α和IL-6基因的表达和分泌。Aim： To investigate the effects of hypoxia on the secretions of proinflammatory cytokines TNF-α and IL-6 and to inquire into the mechanism. Methods： Separated mice abdominal macrophages which were identified with non-specificesterase dye method,and created the hypoxic cultured model. The levels of TNF-α and IL-6 in the medium were determined by ELISA method. The mRNA expressions of TNF-α and IL-6 were measured by RT-PCR method. NF-κB activation was assayed by Western blot method. Finaly, we added cortone（5 μg/ml） to the medium, then observed the secretion levels of TNF-α and IL-6 during hypoxia. Results： The secretions of TNF-α and IL-6 from MФ exposed to hypoxia for 12 h were increased significantly compared with control（ P 〈 0.01 ）. The expressions of TNF-α mRNA and IL-6 mRNA were enchanced obviously contrasted with control（ P 〈 0.01 ）. NF-κB activation in MФ nuclei was raised at 2 h during hypoxia and persisted to 5 h. We added cortone to the medium and found no significant change in secretion of TNF-α and IL-6 during hypoxia. Conclusion： Hypoxia could activate NF-κB and make it shift to nucleus which promoted the transcriptions and expressions of TNF-α and IL-6.

关 键 词：低氧 巨噬细胞 核因子-KAPPA B 肿瘤坏死因子-Α 白介素-6 

分 类 号：Q291[生物学—细胞生物学]