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目的研究体外常氧环境下脂多糖(LPS)诱导小鼠腹腔巨噬细胞缺氧诱导因子-1α(HIF-1α)表达的相关信号通路。方法以LPS单独或联合NF-κB抑制剂柳氮磺胺吡啶(SSZ)、一氧化氮合酶(NOS)抑制剂L-单甲基精氨酸(L-NMMA)作用于BALB/c小鼠腹腔巨噬细胞。用逆转录聚合酶链反应(RT-PCR)法检测HIF-1αmRNA的变化,用免疫细胞化学法检测HIF-1α蛋白的变化,用Griess反应检测培养上清中亚硝酸盐浓度的变化。结果LPS作用于小鼠巨噬细胞10h后,HIF-1αmRNA水平无明显变化(P>0.05),HIF-1α蛋白水平则明显增加(P<0.05)。SSZ和L-NMMA组HIF-1α蛋白的表达则明显低于LPS组(P均<0.05)。与对照组相比,LPS可明显增加巨噬细胞NO的生成(P<0.05),SSZ则可明显抑制LPS诱导的NO的产生(P<0.05)。结论LPS可通过NF-κB及iNOS等转录后信号途径上调HIF-1α的表达。
Objective To investigate the related signal pathway of lipopolysaccharide (LPS) -induced hypoxia inducible factor-1α (HIF-1α) expression in mouse peritoneal macrophages under normoxia in vitro. Methods BALB / c mice were treated with LPS alone or combined with sulfasalazine (SSZ) and L-NMMA, a nitric oxide synthase (NOS) inhibitor. Macrophages. The changes of HIF-1αmRNA were detected by reverse transcriptase-polymerase chain reaction (RT-PCR), the changes of HIF-1αprotein were detected by immunocytochemical method, and the changes of nitrite concentration in culture supernatant were detected by Griess reaction. Results After treated with LPS for 10h, there was no significant change in HIF-1α mRNA level (P> 0.05) and HIF-1α protein level (P <0.05). The expression of HIF-1α in SSZ and L-NMMA group was significantly lower than that in LPS group (all P <0.05). Compared with the control group, LPS significantly increased NO production in macrophages (P <0.05), and SSZ significantly inhibited LPS-induced NO production (P <0.05). Conclusion LPS can up-regulate the expression of HIF-1α through post-transcriptional signaling such as NF-κB and iNOS.