摘要
目的观察内毒素脂多糖(LPS)致伤小鼠肺组织中Src抑制的蛋白激酶C的底物(SSeCKS)的表达变化和细胞定位。方法腹腔注射LPS制备小鼠内毒素肺损伤模型,依据注射时相和剂量不同随机分组。用RealtimePCR法和Westernblot法检测各组肺组织SSeCKS的mRNA和蛋白质水平表达情况,间接免疫荧光双标法检测SSeCKS在肺组织中的细胞定位。结果SSeCKS表达水平与LPS用量呈现剂量和时间依赖关系:1、5、10和15mgkg组SSeCKSmRNA水平皆显著高于对照组(P<0.05),且随注射剂量增高SSeCKSmRNA表达量亦逐渐增高;不同时相LPS(5mgkg)注射后肺组织中SSeCKSmRNA水平表达呈动态变化过程,1h开始增高,3h达到表达高峰,12h降至正常水平;SSeCKS蛋白水平表达变化与mRNA水平变化相一致。SSeCKS在肺组织中定位于肺泡间隔毛细血管内皮细胞。结论SSeCKS是炎症反应蛋白,其表达量与炎症的严重程度相关。内毒素可诱导肺泡间隔毛细血管内皮细胞SSeCKS表达上调。
Obective To investigate the changes of the expression of SSeCKS in the injured pulmonary tissue of mice induced by LPS. Methods The LPS-induced pulmonary tissue injury model of mouse was set up intraperitoneally. The injured mice were grouped randomly according to different dosages of LPS and time phases. The expression of protein as well as the mRNA of SSeCKS in the pulmonary tissue of different groups was determined by Real-time PCR and Western blot, and the location of SSeCKS in the cells of lung tissue was tested by confocal scanning microscopy. Results The levels of SSeCKS expression and the dosages of LPS used were time and dese-dependent. The SSeCKS mRNA levels of groups 1, 5, 10 and 15 mg/kg were significantly higher than those of controls （ P 〈 0.05） and the expression of SSeCKS mRNA was increasing along with the increasing dosages of LPS injected. At different time point after injection of LPS （5 mg/kg）, the expression of SSeCKS mRNA in the pulmonary tissue was of a dynamic process： the expression level started to increase after 1h, reached the peak at 3 h and decreased down to normal one at 12h. And the SSeCKS protein expression was paralleled with the levels of mRNA. SSeCKS was located in the endothelial cells of interalveolar blood capillaries. Conclusion SSeCKS is a kind of reactive protein of inflammation, the expression of which is related with the severity of inflammation. LPS is able to induce the up-regulation of expression of SSeCKS in the endothelial cells of interalvelar blood capillaries.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第3期241-245,共5页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金(30300099)
江苏省自然科学基金(BK2003035)
江苏省高校自然科学基金(03KJB180109)
关键词
LPS
SSECKS
血管内皮细胞
炎症
Lipopolysaccharide （LPS）
Src-suppressed C kinase substrate （SSeCKS）
Vascular endothelial cell
Inflammation
