摘要
目的:研究低氧诱导肺泡巨噬细胞低氧诱导因子-1α(HIF-1α)的表达及HIF-1α对肺泡巨噬细胞 产生肿瘤坏死因子-α(TNF-α)的影响。方法:应用HIF-1α诱骗法(HIF-1α decoy)抑制低氧(3％O2,5％CO2,92％ N2)培养的肺泡巨噬细胞中HIF-1α的作用,并用免疫组织化学、Western blot、半定量RT-PCR、酶联免疫吸附法 (ELISA)分别检测HIF-α蛋白、mRNA的表达和TNF-α的产生。结果:HIF-1α在常氧对照组肺泡巨噬细胞核中表 达呈阴性,在低氧组和HIF-1α decoy组表达呈阳性;低氧组和HIF-1α decoy组中HIF-1α蛋白的含量显著高于常氧 对照组(P<0．05)。HIF-1α mRNA的含量在低氧组和HIF-1α decoy组明显高于常氧对照组(P<0．05);培养的巨噬 细胞上清液中TNF-α的含量在低氧组(115±17 ng／L)明显高于常氧对照组(69±13 ng／L,P<0．05)和HIF-1α decoy 组(8l±15 ng/L,P<0．05)。结论:低氧可明显诱导肺泡巨噬细胞HIF-1α的表达和活性增强,后者能促进TNF-α 的产生,提示在可导致肺部低氧的炎症性疾病如COPD中HIF-1α可能发挥重要作用。
AIM： To study the effect of hypoxia inducible factor - 1 alpha （HIF - 1α） on tumor necrosis factor alpha （TNF - α） production in rat alveolar macrophages cultured under hypoxic condition. METHODS： Using HW - 1α decoy inhibiting its function, Immunohistechemistry, Western blot, semiquantitative RT- PCR and ELISA were used to determine the expression of HIF-1α protein and mRNA and the production of TNF-α in rat alveolar macrophages cultured under hypoxic condition （3% O2, 5% CO2, 92% N2）, respectively. RESULTS： Expression of HIF- 1α was positive in cultured macrophage nucleoli in hypoxia group and HIF-1α decoy group but it was negative in nomoxic control group. The content of HIF - 1α protein in hypoxia group and HIF - 1α decoy group were signiticandy higher than that in nomoxic control group （ P 〈 0.05）. The content of HIF - 1α mRNA in hypoxia group and HIF- 1α decoy group were markedly higher than that in nomoxic control group （ P 〈 0.05）, respectively. The content of TNF -α in hypoxia group （ ll5 ＋ 17 ng/L） was higher than that in control group [ （69 ＋ 13） ng/L, P 〈 0.05] and HIF - la decoy group [（81 ＋ 15） ng/L, P〈0.05]. CONCLUSION： Hypoxia can increase significantly expression and activity of HIF- 1α, which can promote the production of TNF- α in rat alveolar macrophages. It suggests that HIF- 1α plays an important role in the pathogenesis of chronic inflammation- related diseases that can give rise to lung hypoxia such as COPD.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2005年第10期1990-1993,共4页
Chinese Journal of Pathophysiology
基金
湖北省自然科学基金资助项目(2003ABA149)
关键词
低氧
肿瘤坏死因子
巨噬细胞
肺泡
Anoxia
Tumor necrosis factor
Macrophages, alveolar
