苦杏仁甙对高氧暴露早产鼠肺泡Ⅱ型细胞表面活性物质蛋白A、B、C mRNA表达的影响

Effect of amygdalin and hyperoxia on surfactant associated protein A,B,C messenger RNA of type Ⅱ alveolar epithelial cells in premature rat

目的 探讨高氧暴露对体外培养的早产鼠肺泡Ⅱ型细胞 (AECⅡ )表面活性物质蛋白(surfactantassociatedprotein ,SP)A、B、CmRNA表达的影响及苦杏仁甙的作用。 方法 分离、培养19d早产鼠AECⅡ ,经贴壁纯化后随机分为四组 :空气组 (Ⅰ组 )、高氧组 (Ⅱ组 )、空气 +苦杏仁甙组(Ⅲ组 )、高氧 +苦杏仁甙组 (Ⅳ组 )。Ⅱ、Ⅳ组在更换培养液后按 3L/min通入 95 %氧、5 %二氧化碳高纯混合气 ,10min后密封。Ⅲ、Ⅳ组在更换培养液时加入 2 0 0 μmol/L苦杏仁甙 ,培养 2 4h后 ,收获各组细胞 ,Trizol一步法提取细胞总RNA ,RT PCR半定量分析各组培养细胞中SP A、SP B、SP CmRNA表达强度。 结果 Ⅱ组SP A、SP B、SP CmRNA表达强度均较Ⅰ组弱 (P <0 .0 1) ,Ⅲ组与Ⅰ组比较差异无显著性 (P >0 .0 5 ) ,Ⅳ组较Ⅱ组表达强度明显增加 (P <0 .0 5 )。 结论 高氧暴露导致早产鼠AECⅡSP A、SP B、SP CmRNA表达降低 ,这种改变参与了高氧肺损伤的发病过程 ,苦杏仁甙可抑制这种变化 ,提示苦杏仁甙对高氧肺损伤起一定的保护作用。

Objective To explore effect of hyperoxia and amygdalin on surfactant associated protein messenger RNA levels of alveolar epithelial cells of premature rat lung. Methods Type Ⅱ alveolar epithelial cells (AECⅡ) were gained by primary culture from 19-days fetal rat lung. After purified,AECⅡ were randomly assigned to four groups and exposed to air or hyperoxia: air group (group Ⅰ),hyperoxia group (group Ⅱ),air plus amygdalin group (group Ⅲ),hyperoxia plus amygdalin group (group Ⅳ),Groups Ⅱ、Ⅳ were flushed the flake with 95% oxygen-5% CO 2 at 3 L/min for 10 min,then sealed and cultured for 24 hours. Groups Ⅲ,Ⅳ were added 200 μmol/L of amygdalin at the same time. All groups were in CO 2 culture chamber (37 ℃,5% CO 2) for 24 hours,cells were harvested and extracted for total RNA by Trizol reagent. mRNA levels of SP were measured by reverse transcription polymerase chain reaction (RT-PCR). Results SP mRNA levels were significantly decreased in groupⅡ compared to groupⅠ( P <0.01);After amygdalin was added,SP mRNA levels were elevated in group Ⅲ and Ⅳ compared to groupⅡ( P <0.05),but SP mRNA levels were not singnifcantly elevated( P <0.05) compared to groupⅠ. Conclusion Hyperoxia decreased SP mRNAs levels in AECII from premature rat lung might be contributing to hyperoxia-induced lung injury in premature rat;amygdalin could inhibit the changes of SP mRNAs levels of AECII resulted from hyperoxia and may play partial protect role in hyperoxia-induced lung injury.