重组人促红细胞生成素对新生大鼠高体积分数氧肺损伤细胞炎症的影响  

Effect of human recombinant erythropoietin on inflammation of hyperoxic lung injury in neonatal rats

作　　者：黄启丽 任静[2] 谢燕媚[2] 纪泽泉[2] 刘海燕[1] 黄翠雯[1] Huang Qili;Ren Jing;Xie Yanmei;Ji Zequan;Liu Haiyan;Huang Cuiwen(Department of Neonatology,the Second Affiliated Hospital,Guangzhou Medical University,Guangzhou 510000,China;Department of Pediatrics,the Second Affiliated Hospital,Guangzhou Medical University,Guangzhou 510000,China)

机构地区：[1]广州医科大学附属第二医院新生儿科,510000 [2]广州医科大学附属第二医院儿科,510000

出　　处：《中华实用儿科临床杂志》2020年第5期383-388,共6页Chinese Journal of Applied Clinical Pediatrics

基　　金：广东省医学科研基金(A2016073)。

摘　　要：目的探讨人重组促红细胞生成素(rhEPO)对新生大鼠高体积分数氧(高氧)肺损伤炎症的影响。方法将72只新生SD大鼠按随机数字表法分为4组:对照组、支气管肺发育不良(BPD)组、高氧+低剂量促红细胞生成素[EPO(L)]组、高氧+高剂量促红细胞生成素[EPO(H)]组。BPD组、高氧+EPO(L)组和高氧+EPO(H)组新生大鼠暴露于850 mL/L氧气中,高氧+EPO(L)组和高氧+EPO(H)组分别于高氧暴露后1、3、7 d给予800 IU/kg、2000 IU/kg rhEPO皮下注射,对照组和BPD组注射等量9 g/L盐水。实验开始后3、7、14 d记录各组体质量。HE染色,光镜下观察其肺组织形态结构的改变,检测肺组织放射状肺泡计数(RAC)。免疫荧光染色法检测核因子-κB(NF-κB)的表达,Western blot技术检测肺组织磷酸化NF-κB(pNF-κB)、抑制蛋白(IκB)及Caspase-3的表达,酶联免疫吸附试验(ELISA)检测支气管肺泡灌洗液中白细胞介素-1β(IL-1β)的表达。结果1.BPD组新生大鼠14 d时体质量明显低于对照组[(18.97±3.21)g比(27.97±2.30)g],差异有统计学意义(P<0.05);14 d时,高氧+EPO(L)组和高氧+EPO(H)组新生大鼠体质量[(24.16±2.15)g、(26.04±1.97)g]均显著高于BPD组,差异均有统计学意义(均P<0.05)。2.HE染色观察肺组织形态学结构显示,与对照组比较,BPD组3 d肺泡间隔有少量炎性细胞渗出,7 d更为明显,肺泡腔有渗出,14 d出现肺泡数量减少,肺大疱形成,间隔增厚,RAC明显减少(5.50±1.29比14.33±2.80),差异均有统计学意义(P<0.01);高氧+EPO(L)组和高氧+EPO(H)组新生大鼠肺组织病理改变减轻,炎性细胞浸润减少,RAC值在14 d均明显高于BPD组,差异均有统计学意义(均P<0.05)。3.免疫荧光结果显示,BPD组NF-κB p65阳性细胞数目明显增多,荧光强度增强,给予EPO处理后可减少NF-κB p65阳性细胞数目,荧光强度减弱。4.Western blot结果显示,与对照组比较,BPD组pNF-κB p65及Cleaved Caspase-3显著增高,差异均有统计学意义(均P<0.05);IκB明Objective To explore the effects of human recombinant erythropoietin(rhEPO)on inflammation of hyperoxic lung injury in neonatal rats.Methods Seventy-two neonatal rats were randomly divided into 4 groups:control group,BPD group,hyperoxia+low-dose recombinant erythropoietin[EPO(L)]group,and hyperoxia+high-dose recombinant erythropoietin[EPO(H)]group.The neonatal rats in BPD group,hyperoxia+EPO(L)group and hyperoxia+EPO(H)group were exposed to 850 mL/L oxygen.Then the neonatal rats in hyperoxia+EPO(L)group and hyperoxia+EPO(H)group were given 800 IU/kg and 2000 IU/kg rhEPO by subcutaneous injection respectively at 1 d,3 d and 7 d,while the control group and BPD group were given the same amount of 9 g/L saline water.Initially,the body weight of each group was recorded at 3 d,7 d and 14 d.The morphological structure changes of lung tissues were observed by HE staining under light microscope,and the radial alveolar count(RAC)in lung tissues were detected.The expression of nuclear factor kappa B(NF-κB)was detected by immunofluorescence staining;Western blot was applied to determine the protein expression of phosphorylated NF-κB(pNF-κB),inhibitor protein(IκB)and Caspase-3 in lung tissues;and the expression of interleukin-1β(IL-1β)in bronchoalveolar lavage fluid was determined using enzyme linked immunosorbent assay(ELISA).Results(1)On the 14th day,the body weight of neonatal rats in the BPD group was lower than that in the control group[(18.97±3.21)g vs.(27.97±2.30)g],and the difference was statistically significant(P<0.01);however,the body weights of neonatal rats in the hyperoxia+EPO(L)group and hyperoxia+EPO(H)group[(24.16±2.15)g,(26.04±1.97)g]was much heavier than that in the BPD group,and the differences was statistically significant(all P<0.05).(2)The morphological structure of lung tissues which was observed by HE staining showed that in the BPD group,there were a few inflammatory cells infiltration in alveolar septum on the 3rd day,the inflammatory response was more evident on the 7th day,when exudation

关 键 词：支气管肺发育不良 重组人促红细胞生成素 高体积分数氧 炎性反应 核转录因子-ΚB 

分 类 号：R722[医药卫生—儿科]