基础医学与临床 ›› 2013, Vol. 33 ›› Issue (10): 1297-1301.

• 研究论文 • 上一篇    下一篇

LPS预处理增加CoCl2诱导小鼠巨噬细胞表达HIF-1α

江丽丽1,张建斌2,谢平3,谢可鸣4   

  1. 1. 苏州大学基础医学院病理生理学教研室
    2. 连云港市中医院
    3. 无锡人民医院
    4. 苏州大学医学部
  • 收稿日期:2012-12-26 修回日期:2013-03-26 出版日期:2013-10-05 发布日期:2013-09-25
  • 通讯作者: 谢可鸣 E-mail:xkm@suda.edu.cn

Lipopolysaccharide Pre-treatment Increases Expression of HIF-1α in Cobalt chloride-induced murine macrophages

  • Received:2012-12-26 Revised:2013-03-26 Online:2013-10-05 Published:2013-09-25

摘要: 目的 观察LPS预处理对CoCl2诱导的小鼠巨噬细胞表达低氧诱导因子1α(HIF-1α)和血管内皮生长因子(VEGF)的影响。方法 将小鼠巨噬细胞Raw264.7分为对照组、LPS(1mg/L)组、CoCl2 (100μmol/L)组和LPS预处理组(LPS-Pre组)4组并分别置于含相应试剂的培养液中培养,预处理组则先在LPS 浓度为1mg/L 的培养液中预培养8h再转入CoCl2浓度为100μmol/L的培养液培养。RT-PCR检测HIF-1α和VEGF mRNA;免疫细胞化学和Western blot检测HIF-1α和VEGF蛋白。结果 CoCl2组和LPS预处理组Raw264.7表达HIF-1α和VEGF的mRNA和蛋白均高于对照组(p<0.05),其中,后者蛋白表达最高(p<0.05)。结论 LPS预处理能在转录后水平上调CoCl2刺激的巨噬细胞表达HIF-1α和VEGF。

关键词: 关键词:缺氧, 炎症, LPS, CoCl2, HIF-1α

Abstract: Objective To observe the effects of hypoxia simulated by cobalt chloride on the expression of hypoxia-inducible factor-1α(HIF-1α) and vascular endothelial growth factor(VEGF) in murine macrophage Raw264.7 induced beforehand by using LPS . Methods Raw264.7 were divided into control group,LPS(1mg/L)group,CoCl2(100μmol/L)group and LPS pre-treatment(LPS-Pre) group and incubated respectively in the culture containing corresponding reagent. The macrophages of LPS pre-treatment group were at first incubated for 8 hours in the culture with LPS of 1mg/L and then were transferred into the culture containing CoCl2 of 100μmol/L. HIF-1α and VEGF’ mRNA were detected by RT-PCR;protein of HIF-1α and VEGF were analyzed by Western blot and immunocytochemistry. Results Compared with those of control, mRNA and protein expression of both HIF-1α and VEGF can be up-regulated in CoCl2 group and LPS-Pre group(p<0.05).Moreover, cells were pre-treated with LPS and followed activated by CoCl2, HIF-1α’s protein significantly higher(p<0.05)than that of induced only by CoCl2. Conclusion Pre-treatment with LPS can up-regulate HIF-1α and VEGF’s protein at the post-transcriptional level in macrophages under hypoxia environment.

Key words: Key words: hypoxia, inflammation, LPS, CoCl2, HIF-1α

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