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目的了解肺泡巨噬细胞在产生内毒素耐受时,糖皮质激素受体(glucocorticoid receptor,GR)在转录水平的表达。方法培养新生大鼠肺泡巨噬细胞,将细胞随机分为4组(A组为正常对照组,B、C组为小浓度LPS耐受组,D组为高浓度LPS直接刺激组),分别用不同浓度脂多糖(Lipopolysaccharide,LPS)刺激肺泡巨噬细胞24h后,再改变LPS浓度刺激上述各组细胞24h,分别提取RNA和蛋白质,以逆转录聚合酶链反应(RT-PCR)检测GR的mRNA表达。结果GR mRNA与β-actin比值A组高于D组,低于内毒素耐受的B、C组(P<0.01)。D组的表达水平低于内毒素耐受的B、C组(P<0.01)。但内毒素耐受的B、C组间无明显差异。结论内毒素耐受的肺泡巨噬细胞GR表达上调,说明巨噬细胞内毒素耐受时GR起到重要作用。
Objective To investigate the expression of glucocorticoid receptor (GR) at the transcriptional level in endotoxin-tolerant alveolar macrophages. Methods Alveolar macrophages of neonatal rats were cultured and randomly divided into 4 groups (A group was normal control group, B and C group were low concentration LPS tolerance group, D group was high concentration LPS direct stimulation group) After stimulating alveolar macrophages with different concentrations of lipopolysaccharide (LPS) for 24 h, the cells were stimulated by LPS for 24 h, and RNA and protein were extracted respectively. MRNA and protein of GR were detected by reverse transcription-polymerase chain reaction (RT-PCR) expression. Results The ratio of GR mRNA to β-actin in group A was higher than that in group D and lower than those in group B and C with endotoxin tolerance (P <0.01). The expression level of D group was lower than that of endotoxin group B and C (P <0.01). However, there was no significant difference between endotoxin-resistant B and C groups. Conclusion The GR expression of endotoxin-resistant alveolar macrophages is up-regulated, indicating that GR play an important role in the endotoxin tolerance of macrophages.