摘要
目的观察新生大鼠高氧肺损伤模型肺组织中Src抑制的蛋白激酶C的底物(SSeCKS)的表达及细胞定位。方法64只新生大鼠按照随机数字表法分为空气对照组和高氧暴露组,分别建立动物模型,于12、24、48、72h处死动物,进行肺组织的病理学检查及肺湿干质量比(W/D)检测,应用Western blot及免疫组化法检测各组各时相点肺组织中SSeCKS蛋白的表达,用间接免疫荧光双标法明确SSeCKS蛋白在肺组织中的定位。结果病理HE染色:高氧暴露24h肺泡内出现少量红细胞,肺泡壁增厚;48h肺泡内红细胞增多,肺间隔增宽,粒细胞附壁;72h出现肺出血,肺泡内可见大量的红细胞,肺泡间隔粒细胞浸润。Western blot法检测SSeCKS蛋白:空气对照组可检测到极少量的SSeCKS的表达,高氧暴露24h表达增加,72h达到最高,72h内随高氧暴露时间延长而增加;免疫组化定量分析与Western blot结果一致;间接免疫荧光双标法明确SSeCKS定位于毛细血管内皮细胞。结论提示SSeCKS参与了新生大鼠高氧肺损伤的过程。
Objective To investigate the expression of Src-suppressed C kinase substrate (SSeCKS) in the neonatal rat lung after hyperoxia-induced injury. Methods Totally 64 rats at the age of 12 h were randomly and equally divided into 2 groups, controI group (normal air condition) and hyperoxia-induced lung injury group (02 95% and CO2 5% ). Then the rats were executed at the time points of 12, 24, 48 and 72 h after hyperoxic exposure, and the right upper lung was resected for the examination of histopathology and immunohis- tochemistry, the right below lung for wet weight/dry weight ratio, and the left lung for detecting the expression of SSeCKS by Western blot analysis. The location of SSeCKS in the cells of lung tissue was tested by confocal scanning microscopy. Results At 24 h after the hyperoxic exposure, HE staining showed a few red blood cells in the alveoli and thickening alveolar wall. There were more red blood cells, distended alveolar septa and granulocyte infiltration in 48 h. Obvious acute inflammation, including hemorrhage, large amount of red blood cells, inflammatory cell infiltration into alveolar wall appeared at 72 h. In control group, SSeCKS was detected little. But in hyperoxia-induced lung injury group, the expression level of SSeCKS started to increase at 24 h, reached to the peak at 72 h and continued to increase with the elapse of time. SSeCKS was located in the capil- lary endothelial cells of alveolar septa. Conclusion SSeCKS is involved in the process of hyperoxia induced lung injury.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2009年第9期831-834,共4页
Journal of Third Military Medical University
关键词
SSECKS
肺损伤
高氧
新生大鼠
内皮细胞
Src-suppressed C kinase substrate
lung injury
hyperoxia
newborn rats
endothelial cells