高氧诱导下新生大鼠脑损伤的发生机制和前列腺素E1的干预作用

doi:10.13481/j.1671-587x.20190603

摘要/Abstract

摘要： 目的：探讨高氧诱导下新生大鼠脑损伤的发生机制，阐明前列腺素E1（PGE-1）通过内质网应激（ERS）途径发挥保护作用的机制，为高氧所致新生儿脑损伤的治疗提供相关理论依据。方法：90只新生Wistar大鼠随机分为对照组、高氧组和高氧+PGE-1组，每组30只。高氧组和高氧+PGE-1组大鼠放于高氧箱内，对照组大鼠置于同一室内常压条件下。自造模第1天开始，高氧+PGE-1组大鼠腹腔注射PGE-1，另外2组大鼠以同等剂量生理盐水替代。在高氧暴露第1、3和7天，每组随机选取10只大鼠检测体质量和脑组织含水量；HE染色观察3组大鼠脑组织形态表现；TUNEL染色观察3组大鼠脑细胞凋亡情况，计算细胞凋亡指数；Western blotting法检测各组大鼠脑组织中氨基末端蛋白激酶（JNK）和磷酸化JNK （p-JNK）表达量。结果：实验第1、3和7天，与同时间对照组比较，高氧组大鼠体质量明显降低（P<0.05），脑组织含水量增加（P<0.05）；与高氧组比较，高氧+PGE-1组新生大鼠体质量明显升高（P<0.05），脑组织含水量降低（P<0.05）。HE染色，对照组大鼠大脑皮层细胞形态规则，排列有序，极少有炎性细胞的浸润、细胞的水肿；高氧组大鼠脑组织皮层细胞排列紊乱，大脑实质可见炎性细胞浸润，存在脑血管水肿；高氧+PGE-1组大鼠大脑皮层细胞排列整齐，细胞形态相对规则，脑实质炎性细胞数减少，水肿较高氧组减轻。TUNEL染色，对照组大鼠大脑组织以大脑皮层细胞为主，凋亡细胞少见，胞核呈均质蓝色；高氧组大鼠脑组织中胞核呈亮白色，为阳性凋亡细胞。与高氧组比较，同一时间高氧+PGE-1组凋亡细胞数明显减少；高氧组大鼠脑细胞凋亡指数高于高氧+PGE-1组和对照组（P<0.05）。高氧组大鼠脑组织中JNK和p-JNK表达量较对照组明显升高（P<0.05），高氧+PGE-1组大鼠脑组织中JNK和p-JNK表达量较高氧组明显降低（P<0.05）。结论：高氧导致新生大鼠脑损伤的发病机制可能与下调ESR相关p-JNK和JNK蛋白表达有关，且PGE-1对其有保护作用。

关键词: 高氧脑损伤, 内质网应激, 前列腺素E1, 新生大鼠

Abstract: Objective: To investigate the pathogenesis of brain injury induced by hyperoxia in the newborn rats,to elucidate the mechanism of the protect effect of prostaglandin E1 (PGE-1) through endoplasmic reticulum stress(ERS) pathway,and to provide the relevant theoretical basis for the treatment of brain injury of the newborns induced by hyperoxia. Methods: Ninety newborn Wistar rats were randomly divided into control group,hyperoxia group and hyperoxia+PGE-1 group,and there were 30 rats in each group.The rats in hyperoxia group and hyperoxia+PGE-1 group were placed in the hyperoxia box; the rats in control group were placed under the same room pressure conditions.From the 1st day of modeling,the PGE-1 was intraperitoneally injected into the rats in hyperoxia+PGE-1 group.The rats in the other two groups were injected with the same dose of normal saline.On the 1st,3rd and 7th days of hyperoxia exposure,10 rats in each group were randomly selected to detect the body weights and water contents of brain tissue;the morphology of brain tissue of the rats in three groups was observed by HE staining;the apoptosis of brain cells of the rats in three groups was observed by TUNEL staining,and the apopotic indexes were caculated;the expression amount of c-Jun N-terminal protein kinase(JNK) and phosphorylated JNK(p-JNK) proteins in brain tissue of the rats in three groups were detected by Western blotting method. Results: On the 1st,3rd and 7th days,the body weights of the rats in hyperoxia group were significantly lower than those in control group (P<0.05);the water contents in brain tissue of the rats in hyperoxia group were significantly higher than those in control group (P<0.05);compared with hyperoxia group,the body weights of the rats in hyperoxia+PGE-1 group were increased (P<0.05), and the water contents in brain tissue were decreased (P<0.05). The HE staining results showed that the cerebral cortex cells of the rats in control group were regular in shape and arranged in order,with little infiltration of inflammatory cells and edema of cells;the cortical cells of the rats in hyperoxia group were disordered,and the inflammatory cells were found in the brain parenchyma;the cerebral cortex cells of the rats in hyperoxia+PGE-1 group were arranged neatly,the cell morphology was relatively regular,the number of inflammatory cells in brain parenchyma were reduced,and the degree of brain parenchyma edema was less than that in hyperoxic group.The TUNEL staining results showed that the cells in brain tissue of the rats in control group were mainly cerebral cortical cells,and the apoptotic cells were rare and the nucleus showed homogeneous blue;in hyperoxia group,the nucleus was bright white,which were positive apoptotic cells.Compared with hyperoxia group,the number of apoptotic cells in hyperoxia+PGE-1 group was decreased significantly at the same time point.The apoptotic index in brain cells of the rats in hyperoxia group was higher than those in hyperoxia+PGE-1 group and control group (P<0.05).The expression amounts of JNK and p-JNK proteins in brain tissue of the rats in hyperoxia group were significantly higher than those in control group (P<0.05),and the expression amounts of JNK and p-JNK proteins in brain tissue of the rats in hyperoxia+PGE-1 group were significantly lower than those in hyperoxia group (P<0.05). Conclusion: The pathogenesis of brain injury induced by hyperoxia in the newborn rats may be related to the down-regulation of the expressions of ESR-related p-JNK and JNK proteins,and PGE-1 has the protective effect on it.

Key words: hyperoxic brain injury, endoplasmic reticulum stress, prostaglandin E1, newborn rats

中图分类号:

R742.8

王叶, 王红, 朴丽贞, 杨山, 张书剑, 金正勇. 高氧诱导下新生大鼠脑损伤的发生机制和前列腺素E1的干预作用[J]. 吉林大学学报(医学版), 2019, 45(06): 1206-1211.

WANG Ye, WANG Hong, PIAO Lizhen, YANG Shan, ZHANG Shujian, JIN Zhengyong. Pathogenesis of brain injury induced by hyperoxia in newborn rats and intervention of prostaglandin E1[J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1206-1211.

参考文献

[1] 吴瑕,柳国胜,罗瑶,等.细胞粘附因子-1在新生大鼠高氧肺损伤模型肺组织的表达及意义[J].中国新生儿科杂志,2012,27(2): 124-127.
[2] 张喆,曹大伟,张新日.高氧肺损伤机制研究进展[J].国际呼吸杂志,2018,38(6):461-464.
[3] 郑潇玥,陈晓.氧化应激在重症新生儿高氧肺损伤中的作用研究进展[J].南昌大学学报:医学版,2016,56(1):92-95.
[4] NICU早产儿用氧及ROP防治现状调查组.16家三甲医院新生儿重症监护病房早产儿用氧及早产儿视网膜病变防治现状调查[J].中华儿科杂志,2012,50(3): 167-171.
[5] GOREN B,CAKIR A,SEVINC C,et al.Uridine treatment protects against neonatal damage and long-term cognitive deficits caused by hyperoxia[J].Brain Res,2017,1676(12):57-68.
[6] 宋朝敏,王程毅,陈涵强,等.新生大鼠高氧脑损伤的评价研究[J].中国新生儿杂志,2014,29(1):54-57.
[7] ALLI M P,KONTIS D.White matter and congnition in adults who were born preterm[J].PLoS One,2011,6(10):e24525.
[8] VOLPE J J.Brain injury in premature infants:a complex amalgam of destructive and developmental disturbances[J].Lancet Neurol,2009,8(1):110-124.
[9] SEMPLE B D,BLOMGREN K,GIMLIN K,et al.Brain development in rodents and humans:Identifying benchmarks of maturation and vulnerability to injury across species[J].Prog Neurobiol,2013,106-107(7/8):1-16.
[10] 杨磊.胰岛素样生长因子-1在新生大鼠高氧脑损伤的研究进展[J].吉林医学,2016,37(3):700-702.
[11] VOTTIER G,PHAM H,PANSIOT J,et al.Deleterious effect of hyperoxia at birth on white matter damage in the newborn rat[J].Dev Neurosci,2011,33(3-4):261-269.
[12] KIM I,XU W,REED J C,et al.Cell death and endoplasmic reticulum Stress:disease relevance and therapeutic opportunities[J].Nat Rev Drug Discov,2008,7(12):1013-1030.
[13] 杨山,张有辰,李慧文,等.前列腺素E1对高氧诱导新生大鼠脑损伤额保护作用[J].中国当代儿科杂志,2018,20(3):230-235.
[14] 黄久浪,韩颖,崔晨,等.内质网应激在高氧导致新生小鼠脑白质损伤中的调控作用[J].中国新生儿在职,2016,31(3):206-211.
[15] 刘春云,刘永平,龚享文.前列地尔药理学研究进展及在呼吸系统疾病中的应用[J].中国新药与临床杂志,2012,31(11):648-651.
[16] MATSUMOTO A,OKIURA T,MORIMATSU F,et al.Effects ofhyperbaric exposure with high oxygen concentration on thephysical activity of developing rats[J].Dev Neurosci,2007,29(6): 452-459.
[17] GERSTNER B,DESILVA TM,GENZ K,et al.Hyperoxia CausesMaturation-dependent cell death in the developing white matter[J].J Neurosci,2008,28(5): 1236-1245.
[18] BHARDWAJ M,PAUL S,JAKHAR R,et al.Potential role of vitexin in alleviating heat stress-induced cytotoxicity:Regulatory effect of Hsp90 on ER stress-mediated autophagy[J].Life Sci,2015,142(12):36-48.
[19] URANO F,WANG X.BERTOLOTTI A,et al.Coupling of stress in the ER to activation of JNK protein kinases by transmembrane protein kinase IRE 1[J].Science,2000,287(5453):664-666.
[20] SHIGEMI Z,MANABE K,HARA N,et al.Methyseleninic acid and sodium selenite induce severe ER stress and subsequent apoptosis through UPR activation in PEL cells[J].Chem Biol Interact,2017,266(3):28-37.
[21] 王一超,刘俊杰,刘海宁,等. Toll样受体4在蛛网膜下腔出血大鼠早期脑损伤中的作用及对海马区神经元自噬的影响[J/OL].西安交通大学学报:医学版,2019[2019-11-18]. http://kns.cnki.net/kcms/detail/61.1399.r.20190930.1115.006.html.