摘要
目的探讨肺组织水通道蛋白5(AQP5)、血及肺灌洗液Clara细胞蛋白(CC16)在急性肺损伤早期的动态改变及临床意义。方法将140只SD新生大鼠随机分两批(每批70只),再随机分为正常对照组(NS组)及内毒素(LPS)组(LPS组,7个亚组)。LPS组予腹腔注射5 mg/kg LPS制作新生大鼠急性肺损伤(ALI)模型。各亚组分别在注射后0.5、1、2、4、8、16 h和24 h处死后取材,第一批收集肺行组织病理和肺湿/干重比值检测,采集血清和支气管肺泡灌洗液(BALF),用双抗体夹心ELISA法检测CC16含量。第二批取肺组织,用免疫印迹法测肺组织AQP5的表达,观察其动态改变。结果①LPS注射0.5h后在病理上动物出现点状肺出血,随时间推延渐发展为片状肺出血;LPS 2 h、4 h组肺湿/干重比(W/D)明显增高,p<0.01有统计学意义;②LPS 0.5 h组大鼠肺组织AQP5表达开始下调,1～2 h组达到最低,继而AQP5表达渐上升,8h组与正常水平接近,之后再次下调至24 h。③LPS致伤后大鼠BALF中CC16水平迅速下降,LPS组各组均较对照组有显著性差异(p<0.01);LPS组血清CC16水平较对照组上升,从注射LPS 2 h后开始有差异,4 h达高峰,以后呈下降趋势,但8 h、16 h组仍明显高于对照组(p<0.01)。结论①新生大鼠AQP5、CC16的改变反映了Ⅰ型肺泡上皮细胞受损及Clara细胞损伤,三者可作为早期反映ALI的敏感实验室指标。②LPS组AQP5、BALF及血清CC16与对照组有差异的时间点有差别,早期发现干预ALI对前两者监测更好。③AQP5表达变化提示可能存在治疗窗口期。
Objective To discuss Aquaporin 5 （AQP5）and CC16 change and clinical significance in early stage of acute lung injury in neonatal rats. Methods One hundred and forty newborn rats were randomized into two group （ every group n = 70 ）, subgroup rats were randomized into normal saline pretreated group （ NS group） and LPS group. Lung injury models in neonatal rats were established via intraperitoneal injection of 5 mg/kg lipopolysaechar- ides （ LPS）, The pathological changes were observed under microscopy, Wet/dry ratio of lung were examined. The ex- pression of AQP5 was examined by Western Blot. Clara cell secretory protein （CC16） in serum and BALF was detec- ted using enzyme - linked immunoadsordent assay（ELISA）. Results① Half - hour following LPS administration to neonatal rats,lung hemorrage was observed, these changes aggravated with time after LPS administration. Wet/dry ratio in neonatal rats at 2 -4 hours after LPS were significantly increased, p 〈0. 01 ,then decreased.② 0. 5 hour after injecting LPS, AQP5 level of lung began descending, it became the lowest after 1 - 2 hours, Significant differences were observed in AQP5 between the NS and LPS pretreated groups（p 〈0. 01 ）. then the level became escalate and approached normal at 8 hours, aquaporin -5 level went down again to 24 hours（p 〈0. 01 ）. ③The contents of CC16 in BALF were significant lower after the injection of LPS, Significant differences were observed in the contents of CC16 in BALF between the NS and LPS pretreated groups（p 〈 0. 01 ）. The contents of CC16 in serum were higher level compared to NS group（p 〈0. 01 ）, In LPS groups CC16 expression peaked at 4 h,8 andl6 hours decreased. Conclu- sion ① The change of AQP5 reflect damage in type I epithelial cells. The dynamic changes of CC16 showed the Clara cells and the integrity of alveolar epithelium and vascular endothelial cell injury. These three is a sensitive marker reflecting in early stage of ALI. ② The pront of time is difference in AQP5 ,CC16 in BALF and serum. In orth- er to early discover and intervent ALI, it is preferably to monitor the former two. ③Expresstion of AQP5 suggests therapeutic window period.
出处
《现代医院》
2013年第1期17-20,共4页
Modern Hospitals
基金
广东省中医药局科研课题(编号:2010207)
