LPS预处理增加CoCl_2诱导小鼠巨噬细胞表达HIF-1α

Lipopolysaccharide pre-treatment increases expression of HIF-1α in cobalt chloride-induced murine macrophages

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(1 mg/L)组、CoCl2(100μmol/L)组和LPS预处理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1 mg/L的培养液中预培养8 h再转入CoCl2浓度为100μmol/L的培养液中培养。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,预处理组蛋白表达最高(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。

Abstract： Objective To observe the effects of hypoxia caused by cobalt chloride to the culture 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 （ 1 mg/L） group,CoCl2 （100 t^mol/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 1 mg/L and then were transferred into the culture containing CoC12 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 im- munocytochemistry. Results mRNA and protein expression of both HIF-1α and VEGF can be up-regulated in CoC12 group and LPS-Pre group（ P 〈 0. 05 ）. The level of HIF--1α protein expression in LPS-Pre group was signifi- cantly higher（P 〈 0.05）than that of the group induced only with CoC12. Conclusions Pre-treatment with LPS can up-regulate environment protein expressions HIF-1α and VEGF at the post-transcriptional level in macrophages under hypoxia