GMCSF对高氧暴露新生大鼠肺组织RAGE—NF—κB通路的影响  

Protection of hyperoxia-induced lung injury by granulocyte-macrophage colony-stimulating factor via RAGE-NF-κB signaling pathway in newborn rats

作　　者：田兆方[1] 张志敏[1] 李玉红[1] 赵赛[1] 王祥[1] 

机构地区：[1]南京医科大学附属淮安第一医院新生儿科,江苏淮安223300

出　　处：《中华医学杂志》2011年第30期2143-2147,共5页National Medical Journal of China

摘　　要：目的探讨高氧暴露对新生大鼠肺组织高级糖基化终产物受体（RAGE）一核因子κB（NF-κB）信号通路的影响及粒细胞巨噬细胞集落刺激因子（GMCSF）肺损伤保护作用的相关机制。方法24只3日龄新生大鼠随机平均分为3组：正常对照组（空气环境下7d）、高氧对照组（95％氧暴露7d）、高氧干预组（95％氧暴露7d＋GMCSF皮下注射9μg／kg／次，共3次）；光镜观察并盲法进行肺组织病理学损伤评分；RT-PCR法检测肺组织匀浆RAGEmRNA、NF—κB mRNA表达；Western印迹检测肺组织匀浆RAGE、NF-κB蛋白表达；ELISA检测支气管肺泡灌洗液（BALF）及血清中肿瘤坏死因子-α（TNF-α）水平；免疫组化法检测肺组织石蜡切片RAGE表达情况。结果正常对照组、高氧对照组、高氧干预组肺组织损伤评分分别为0．46±0．20、3．06±0．33、2．31±0．56，差异有统计学意义（P=0．000）；3组RAGE mRNA和蛋白表达分别为0．14±0．02、0．34±0．06、0．28±0．04和0．30±0．04、0．76±0．11、0．55±0．08，差异均有统计学意义（均P=0．000）；3组NF-κBmRNA和蛋白表达分别为0．41±0．21、0．90±0．36、0．69±0．30和0．41±0．26、0．96±0．43、0．77±0．33，差异均有统计学意义（P=0．000和P=0．017）；3组BALF中TNF—α水平分别为76±10、224±42、143±24，差异有统计学意义（P=0．000）。以上各指标结果在高氧对照组、高氧干预组均明显高于正常对照组（均P〈0．05），高氧干预组低于高氧对照组（P〈0．05）。3组血清TNF-α水平差异无统计学意义（P〉0．05）。结论GMCSF可能通过下调RAGE—NF-κB信号通路对高氧肺损伤发挥保护作用。Objective To explore the effects of granulocyte-macrophage colony-stimulating factor （GMCSF） on hyperoxia exposure lung injury in newborn rats and elucidate its protective mechanism of operating via the signaling pathway of advanced glycation end products （RAGE） -NF-κB. Methods Twenty- four 3-day-old SD rats from 3 litters were randomly divided into 3 groups. They were hyperoxia exposure plus GMCSF group （group A）, hyperoxia exposure group （group B） and air exposure group （group C）. The rats from groups A and B were placed in a sealed Plexiglas chamber with a minimal in-and-outflow, providing 6- 7 exchanges per hour of chamber volume and maintaining 02 levels above 95%. While the rats in group C only were exposed to air simultaneously. The rats in group A received subcutaneous injections of recombinant murine GMCSF （9 μg/kg） during hyperoxia exposure at 24 h, 72 h and 120 h respectively. And the rats in groups B and C received subcutaneous injections of saline vehicle alone at the same time point. Seven days later, all were sacrificed and immunohistochemistry was employed to assess the expression of RAGE in lung tissue. The levels of tumor necrosis factor-or in bronchoalveolar lavage fluid （BALF） and serum samples were detected by ELISA （enzyme-linked immunosorbent assay）. The RAGE mRNA and NF-κB mRNA in tissue homogenates were detected by RT-PCR while RAGE and NF-κB by Western blot. Also the values of lung damage score were calculated with microscopic histology. Results The value of lung damage score in group C, B and A was 0. 46 ± 0. 20, 3.06 ± 0. 33 and 2. 31 ± 0. 56 respectively, there was significantly difference among three groups （ P = 0. 000）. The expression of RAGE mRNA and protein in three groups were 0. 14 ± 0. 02, 0. 34± 0. 06, 0. 28 ± 0. 04 and 0. 30 ± 0. 04, 0. 76 ± 0. 11, 0. 55± 0. 08 respectively. There were both significantly differences among three groups （P = 0. 000, P = 0. 000 ）. The expression of NF-κB mRNA and protein in three groups were

关 键 词：高氧症 肺 大鼠 粒细胞巨噬细胞集落刺激因子 糖基化终产物 高级 

分 类 号：R722[医药卫生—儿科]