高氧、维甲酸对胎鼠肺角化细胞生长因子及其受体表达的影响

The effect of retinoic acid on the expression of keratinocyte growth factor and keratinocyte growth factor receptor in hyperoxia in rat embryonic lung fibroblasts and type II alveolar epithelial cell

目的 探讨高氧、维甲酸 (RA)对胎鼠肺成纤维细胞 (LFs)和肺泡II型上皮细胞 (AECII)角化细胞生长因子 (KGF)及其受体 (KGFR)表达的影响。方法 原代培养胎鼠肺LFs及AECII ,待生长至亚汇合状态时 ,随机分为 :空气组 ,空气 +RA组 ,高氧组 ,高氧 +RA组。于培养 2、6、12、2 4、4 8(AECII)和 72h(LFs)时 ,采用半定量RT -PCR方法检测KGF和KGFR的mRNA表达。结果 与空气组比较 ,高氧 2h时 ,胎鼠LFsKGFmRNA表达即明显下降 ,6h时达极点 ,以后下降趋势逐渐减弱 ,至 4 8h后已无显著性差异 ;高氧 2、6、12和 2 4h时 ,其下降幅度分别为 3 0、5 2、2 3和 2 1倍 (P <0 0 5、0 0 1、0 0 1和 0 0 5 )。RA可上调高氧状态下胎鼠LFsKGFmRNA表达 ,与高氧组比较 ,高氧 +RA组在 2、6、12和 2 4h时KGFmRNA表达分别是高氧组的 2 4、3 4、1 7和 1 8倍 (P <0 0 5、0 0 1、0 0 1和 0 0 5 )。高氧对胎鼠AECIIKGFRmRNA表达影响较小 ,与空气组比较 ,仅在高氧 12h和 2 4h时其表达量分别是空气组的 2 3和 1 3倍 (P <0 0 5 ) ;RA对AECIIKGFRmRNA表达没有影响。

Objective To study the effect of retinoic acid on the expression of keratinocyte growth factor (KGF) and keratinocyte growth factor receptor (KGFR) in hyperoxia in rat embryonic lung fibroblasts (LFs) and type II alveolar epithelial cells (AEC II).Methods The primary rat embryonic LFs and AEC II were cultured in vitro.Cells grew to subconfluence and then were randomly divided into 4 groups: I.Air group; II.Hyperoxia group; III.Air plus RA group; IV.Hyperoxia plus RA group.After cells were cultured for 2,6,12,24,48 (AEC II) and 72 h (LFs),the levels of KGF and KGFR mRNA were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).Results Compared with group I,after hyperoxia,the level of KGF mRNA in rat embryonic LFs decreased at 2 h significantly,with a maximum at 6h,and no significant differences after 48 h.There were 3.0,5.2,2.3 and 2.1 fold decrease at 2,6,12 and 24 h respectively(P<0.05、0.01、0.01 and 0.05).RA could up-regulate the expression of KGF mRNA remarkably in hyperoxia.Compared with group II,there were 2.4,3.4,1.8 and 1.7 fold increase in group Ⅳ at 2,6,12 and 24h respectively(P<0.05、0.01、0.01 and 0.05).Hyperoxia had a modest effect on the expression of AEC II KGFR mRNA.Compared with group I after hyperoxia,the level of KGFR mRNA increased 2.3 and 1.3 fold only at 12 and 24h respectively (P< 0.05).RA had no effect on the expression of AEC II KGFR mRNA.Conclusion RA had a significant protective/preventive effect against hyperoxia lung injury through increasing the expression of KGF.