4-羟基-2,2,6,6-四甲基哌啶对间歇缺氧早产大鼠肺损伤的保护作用  

Protective effect of 4-hydroxy-2,2,6,6-tetramethylpiperidine on lung injury with intermittent hypoxia in premature rats

作　　者：王娟梅[1] 何少茹[2] 张爱民[1] 李云[1] Wang Juanmei;He Shaoru;Zhang Aimin;Li Yun(Department of Pediatrics,the First Affiliated Hospital of Hunan Normal University,Hunan Provincial People′s Hospital,Hunan Provincial Key Laboratory of Pediatric Respirology,Changsha 410000,China;Department of Pediatrics,Guangdong Provincial People′s Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China)

机构地区：[1]湖南省人民医院(湖南师范大学附属第一医院)儿科,儿童呼吸病学湖南省重点实验室,长沙410000 [2]广东省人民医院/广东省医学科学院儿科,广州510080

出　　处：《中华实用儿科临床杂志》2022年第13期1017-1022,共6页Chinese Journal of Applied Clinical Pediatrics

基　　金：湖南省教育厅科学研究项目(16C0972);湖南省人民医院博士基金(BSJJ202013)。

摘　　要：目的了解4-羟基-2,2,6,6-四甲基哌啶(Tempol)对低体积分数给氧干预下间歇缺氧早产大鼠肺组织缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)的表达及肺发育的影响。方法建立给氧间歇缺氧模型。于孕21 d估计胎鼠临产剖宫取鼠,存活早产大鼠192只按照随机数字表法分为6组:空气对照+盐水组、空气对照+Tempol组、恒定给氧+盐水组、恒定给氧+Tempol组、给氧间歇缺氧+盐水组、给氧间歇缺氧+Tempol组,每组32只。于出生7、14、21 d,每组各取8只采用化学分析法检测早产大鼠肺组织丙二醛(MDA)、总抗氧化能力(TAOC),采用实时荧光定量PCR(qPCR)和免疫组织化学法分别检测HIF-1α和VEGF的mRNA和蛋白水平;每组另取8只21 d大鼠行肺功能检测。多组比较采用单因素方差分析,两两比较采用SNK-q检验。结果给氧间歇缺氧+盐水组较恒定给氧+盐水组早产鼠出现轻度肺间隔增厚,MDA增加,TAOC下降,VEGF、HIF-1αmRNA和蛋白表达上升,肺功能指数下降,差异均有统计学意义(均P<0.05);给氧间歇缺氧+Tempol组较相应盐水组MDA下降、TAOC上升[14 d MDA(3.09±0.45)nmol/(mg·pr)比(4.02±0.30)nmol/(mg·pr)、TAOC(3.13±0.31)U/(mg·pr)比(2.44±0.22)U/(mg·pr),21 d MDA(2.87±0.43)nmol/(mg·pr)比(4.47±0.56)nmol/(mg·pr)、TAOC(3.47±0.35)U/(mg·pr)比(2.31±0.32)U/(mg·pr)],差异均有统计学意义(均P<0.05);给氧间歇缺氧+Tempol组较相应盐水组,HIF-1α、VEGF mRNA和蛋白表达下降,于14 d HIF-1αmRNA(2.11±0.60比2.88±0.59)下降的差异有统计学意义(P<0.05),肺功能指数潮气量[(0.41±0.01)mL比(0.36±0.02)mL]、每分钟呼吸通气量[(35.48±2.95)mL比(30.62±2.27)mL]、最大呼气流量[(2.19±0.19)mL/s比(1.51±0.19)mL/s]、动态肺顺应性[(2.65±0.40)mL/cmH2O比(1.83±0.34)mL/cmH2O,1 cmH2O=0.098 kPa]均增加(均P<0.05)。结论Tempol可减轻低体积分数给氧干预下间歇缺氧所致的早产大鼠氧化应激肺损伤,并改善其肺功能。Objective To investigate the effects of 4-hydroxy-2,2,6,6-tetramethylpiperidine(Tempol)on the expressions of hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)and lung development in premature neonatal rats under intermittent hypoxia(achieved by supplying a low concentration of oxygen).Methods The intermittent hypoxia model was established.Caesarean section of rats was performed at 21 days of gestation when the fetal rats were estimated to be in labor.A total of 192 premature neonatal rats survived and were randomly divided into 6 groups according to random number table method:air control+saline group,air control+Tempol group,constant oxygen+saline group,constant oxygen+Tempol group,intermittent hypoxia+saline group,and intermittent hypoxia+Tempol group,32 rats in each group.On the 7th,14th and 21st day of birth,the lung tissues of 8 neonatal preterm rats in each group were taken.Malondialdehyde(MDA)and total antioxidant capacity(TAOC)were detected by chemical analysis.The mRNA and protein levels of HIF-1αand VEGF were detected by real-time fluorescence quantitative PCR(qPCR)and immunohistochemistry,respectively.Another 8 neonatal rats in each group were taken for pulmonary function test on the 21st day after birth.One-way ANOVA and SNK-q test were used for comparison among and between groups,respectively.Results Compared with the constant oxygen+saline group,the intermittent hypoxia+saline group showed mild pulmonary septal thickening,increased MDA,decreased TAOC,elevated mRNA and protein expression levels of VEGF and HIF-1α,and decreased lung function indexes.The differences were statistically significant(all P<0.05).Compared with the corresponding saline group,the intermittent hypoxia+Tempol group had decreased MDA and increased TAOC,and the differences were statistically significant at 14 d[MDA(3.09±0.45)nmol/(mg·pr)vs.4.02±0.30)nmol/(mg·pr),TAOC(3.13±0.31)U/(mg·pr)vs.(2.44±0.22)U/(mg·pr)]and 21 d[MDA(2.87±0.43)nmol/(mg·pr)vs.(4.47±0.56)nmol/(mg·pr),TAOC(3.47±0.35)U/

关 键 词：早产 大鼠 肺发育 间歇缺氧 缺氧诱导因子-1Α 血管内皮生长因子 

分 类 号：R965[医药卫生—药理学]