摘要
目的探讨水通道蛋白1、5(AQP1、AQP5)在脂多糖(LPS)诱导大鼠急性肺损伤(ALI)组织中的表达变化及意义。方法将Wistar大鼠48只随机分为对照组(NC)和急性肺损伤(ALI)组,ALI组通过尾静脉注射脂多糖(LPS),分别于2、6、12和24 h采集标本。HE染色观察肺组织病理;测定肺湿/干重比(W/D)、肺通透指数;ELISA检测血清肿瘤坏死因子α(TNF-α)、巨噬细胞炎性蛋白1α(MIP-1α)水平;Western blot、免疫组化和real-time PCR法检测肺AQP1、AQP5蛋白和mRNA变化。结果与对照组比较,ALI组TNF-α、MIP-1α均有显著性升高(P<0.05),至24 h点逐渐恢复。注射LPS后2 h出现肺泡和间质水肿,炎性细胞浸润,以12 h最明显;肺W/D比、肺泡灌洗(BALF)蛋白含量、肺通透指数均较对照组明显升高(P<0.05),12 h点达峰值。ALI组各时间点肺AQP1及AQP5mRNA及蛋白表达均较对照组明显下调(P<0.01),以12 h点下降最明显。肺AQP1、AQP5 mRNA表达与肺W/D比、血清TNF-α,MIP-1α存在明显负相关。结论脂多糖诱导大鼠急性肺损伤,其肺水肿的形成可能与AQP1、AQP5的表达下调有关。
Objective To investigate the expression and significance of the Aquaporin（ AQP） 1,5 in the LPSinduced lung injury in rats. Methods Totally 48 Wistar rats were divided into control-group,ALI-group. The acute lung injury（ ALI） model was developed by intravenous injection of LPS in rats,control-group was injected with saline. Rats were sacrificed in 2,6,12 and 24 h,and the samples were collected. Observing lung HE staining morphological,determining lung wet / dry weight ratio（ W / D） and lung permeability index,ELISA detecting serum TNF alpha,MIP-1 alpha level. At the same time,analysis AQP1,AQP5 protein and mRNA content changes with Western blot,Immunohistochemistry and real-time PCR method. Results Compared with controlgroup,the TNF alpha,MIP-1 alpha in ALI-group were significantly increased at 2,6 and 12 h（ P〈0. 05）,and to 24 h point gradually down to normal level. HE staining showed that the alveolar and interstitial edema and inflammatory cell infiltration occured 2 h after LPS injection,most obviously in 12 h. The lung W / D,BALF protein and pulmonary permeability index in ALI-group were significantly higher than those of the control-group（ P〈0. 05）,and reached the peak in 12 h. At the same time,the expression level of AQP1,AQP5 in ALI-group was significantly lower than that in control-group（ P〈0. 01）. The correlation analysis showed that the expression of AQP1 and AQP5 mRNA was negatively correlated with the lung W / D ratio,serum TNF alpha and MIP-1 alpha level.Conclusions LPS can induce rats acute lung injury. The formation of pulmonary edema may be related to the down-regulation of AQP1 and AQP5.
出处
《基础医学与临床》
CSCD
2016年第1期53-57,共5页
Basic and Clinical Medicine
基金
福建省医学创新课题基金(2012-CXB-28)
关键词
水通道蛋白
肺损伤
脂多糖
大鼠
aquaporins
lung injury
lipopolysaccharide
rats
