角质细胞生长因子-2对ALI大鼠肺组织趋化因子FKN及紧密连接蛋白claudin-5表达的影响  

Effects of keratinocyte growth factor-2 on expressions of chemokine FKN and tight junction protein claudin-5 in lung of rats with acute lung injury

作　　者：邵腾皓 白吉佳 马希刚[2] Shao Tenghao;Bai Jijia;Ma Xigang(Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Department of Intensive Care Unit,Ningxia Medical University General Hospital,Yinchuan 750004,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏医科大学,宁夏回族自治区银川750004 [2]宁夏医科大学总医院重症医学科,宁夏回族自治区银川750004

出　　处：《中华危重病急救医学》2018年第11期1066-1070,共5页Chinese Critical Care Medicine

基　　金：宁夏回族自治区自然科学基金(NZ17151).

摘　　要：目的探讨角质细胞生长因子-2(KGF-2)对急性肺损伤(ALI)大鼠肺组织趋化因子FKN和紧密连接蛋白claudin-5表达的影响。方法以健康雄性SD大鼠为研究对象,按随机数字表法分为生理盐水(NS)对照组、ALI模型组和KGF-2预处理组,每组10只。经尾静脉注射0.01mL/kg油酸建立大鼠ALI模型;NS对照组注射等量NS。KGF-2预处理组于制模前72h经大鼠气道内滴注5mg/kg KGF-2;NS对照组及ALI模型组滴注等量NS。各组于制模后8h取大鼠腹主动脉血,处死大鼠后对左肺支气管肺泡进行灌洗,收集支气管肺泡灌洗液(BALF),测定血浆和BALF中蛋白水平,计算肺通透性指数(LPI)。取右肺组织,用苏木素-伊红(HE)染色,光镜下观察其病理学改变,并计算ALI病理学评分(LIS);测定肺湿/干重(W/D)比值;采用免疫组化法及蛋白质免疫印迹试验(Western Blot)对肺组织中FKN和claudin-5的表达进行定性、定量分析。双变量相关性采用直线或曲线拟合相关分析。结果ALI模型组大鼠肺组织损伤严重,出现明显的病理学改变,表现为肺泡间隔增厚,炎性细胞浸润;LIS评分、肺W/D比值及LPI均较NS对照组明显升高[LIS(分):3.56±0.28比0.62±0.36,肺W/D比值:6.37±0.29比4.39±0.33,LPI:3.46±0.69比0.98±0.17,均P<0.01]。与NS对照组比较,ALI模型组肺组织FKN阳性表达明显增强,表达量明显升高(FKN/GAPDH:0.97±0.18比0.62±0.04,P<0.01);claudin-5阳性表达明显减弱,表达量明显降低(claudin-5/GAPDH:0.56±0.11比1.06±0.13,P<0.01);FKN与claudin-5蛋白表达量呈显著负相关(r=-0.817,P=0.025)。给予KGF-2预处理后,大鼠肺组织损伤程度明显减轻,病理学改变明显改善;LIS评分、肺W/D比值及LPI均较ALI模型组明显降低[LIS(分):2.41±0.17比3.56±0.28,肺W/D比值:5.45±0.55比6.37±0.29,LPI:2.42±0.19比3.46±0.69,均P<0.01]。与ALI模型组比较,KGF-2预处理组大鼠肺组织FKN阳性表达明显减弱,表达量明显下降(FKN/GAPDH:0.79±0.03比0.97±0.18,P<0.01);而claudiObjective To observe the effects of keratinocyte growth factor-2(KGF-2)on the expressions of chemokine FKN and tight junction protein claudin-5 in lung tissue of rats with acute lung injury (ALI). Methods Healthy male Sprague-Dawley (SD)rats were randomly divided into normal saline (NS)control group,ALI model group and KGF-2 pretreatment group,with 10 rats in each group.The rat ALI model was reproduced by injection of 0.01mL/kg oleic acid into the tail vein,and the rats in NS control group were injected with the same amount of NS. The rats in KGF-2 pretreatment group were instilled with 5mg/kg KGF-2 in the airway at 72 hours before the model reproduction,and the rats in the NS control group and the ALI model group were instilled with the same amount of NS. The abdominal aortic blood of rats was collected at 8 hours after model reproduction,and then the rats were sacrificed, bronchoalveolar in left lung was lavage,and the bronchoalveolar lavage fluid (BALF)was collected for determination of protein levels in plasma and BALF,and the lung permeability index (LPI)was calculated.The lung tissues were harvested, after hematoxylin-eosin (HE)staining,the histopathological changes were observed under light microscope,and the ALI pathology score (LIS)was calculated.The lung wet/dry weight (W/D)ratio was determined.Immunohistochemistry and Western Blot were used to qualitatively and quantitatively analyze the expressions of FKN and claudin-5in the lung tissue.The correlation between two variables was analyzed by linear or curve fitting correlation analysis.Results In the ALI model group,the lung tissue was severely damaged,and obvious pathological changes were observed,including thickened alveolar space and inflammatory cell infiltration,and LIS score,lung W/D ratio and LPI were significantly higher than those of the NS control group (LIS:3.56±0.28vs.0.62±0.36,lung W/D ratio:6.37±0.29vs.4.39±0.33, LPI:3.46±0.69vs.0.98±0.17,all P <0.01).Compared with the NS control group,the positive expression of FKN in the lung tissu

关 键 词：急性肺损伤 趋化因子FKN 紧密连接蛋白claudin-5 角质细胞生长因子-2 

分 类 号：R563[医药卫生—呼吸系统]