促红细胞生成素对高氧脑损伤新生大鼠CCAAT增强子结合蛋白同源蛋白及葡萄糖调节蛋白78表达的影响
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摘要
[目的]探讨促红细胞生成素(EPO)对高氧脑损伤新生大鼠CCAAT增强子结合蛋白同源蛋白(CHOP)及葡萄糖调节蛋白78(GRP78)表达的影响.[方法]取足月新生Wistar大鼠随机分为正常对照组(FiO_2为21%)、高氧组(FiO_2高于85%)和高氧+EPO组(FiO_2高于85%).将正常对照组新生大鼠置于室内空气中饲养,高氧组和高氧+EPO组大鼠置于FiO_2高于85%的室内玻璃箱中饲养.实验第1 d起,高氧+EPO组大鼠每日腹腔注射给予EPO(800 U/kg),实验第1,3,7,14 d时测定各组大鼠脑组织湿/干质量(W/D)比值,采用蛋白免疫印记法检测脑组织中CHOP,GRP78蛋白表达水平.[结果]高氧组新生大鼠第1,3,7,14 d时的脑组织W/D值较正常对照组显著升高(P<0.05),高氧下暴露时间越长,W/D值升高越显著;高氧+EPO组新生大鼠脑组织含水量较高氧组显著降低(P<0.05).高氧组第1,3,7,14 d时的CHOP和GRP78蛋白表达水平较正常对照组显著升高(P<0.05),高氧+EPO组CHOP,GRP78蛋白表达水平较高氧组显著降低(P<0.05).[结论] EPO可缓解高氧引起的新生大鼠脑水肿,对高氧脑损伤具有神经保护作用,其机制认为可能与下调CHOP和GRP78蛋白表达有关系.
OBJECTIVE To investigate the effects of erythropoietin(EPO) on the expressions CCAAT enhancer binding protein homologous protein(CHOP) and glucose regulated protein 78(GRP78) in neonatal rats with hyperoxic brain injury. METHODS Full-term newborn Wistar rats were randomly divided into control group(FiO_2=21%), hyperoxia group(FiO_2>85%) and hyperoxia + EPO group(FiO_2>85%). The newborn rats in the control group were placed in indoor air, and the rats in the hyperoxia and the hyperoxia+EPO group were placed in an indoor glass box with FiO_2 more than 85%. From the first day of the experiment, the rats in the hyperoxia + EPO group were intraperitoneally injected with EPO(800 U/kg), and the wet/dry mass(W/D) ratio of brain tissue was measured at day 1, 3, 7 and 14. Protein immunoblotting was used to detect the expression levels of CHOP and GRP78 protein in brain tissue. RESULTS The W/D values of brain tissue on the 1st, 3rd, 7th and 14th day of neonatal rats in the hyperoxia group were higher than those in the control group(P<0.05). The longer the exposure time under high oxygen, the more obvious the increase of W/D values. The water content of neonatal rat brain tissue in the hyperoxia + EPO group was obviously lower than that in the hyperoxia group(P<0.05). The expressions of CHOP and GRP78 protein in the hyperoxia group were significantly higher than those in the control group at day 1, 3, 7 and 14(P<0.05). The expression of CHOP and GRP78 protein in the hyperoxia + EPO group was significantly lower than those in the hyperoxia group(P<0.05). CONCLUSION EPO could alleviate cerebral edema induced by hyperoxia in neonatal rats and had neuroprotective effects on hyperoxic brain injury, which might be related to the down-regulation of CHOP and GRP78 protein expression.
OBJECTIVE To investigate the effects of erythropoietin(EPO) on the expressions CCAAT enhancer binding protein homologous protein(CHOP) and glucose regulated protein 78(GRP78) in neonatal rats with hyperoxic brain injury. METHODS Full-term newborn Wistar rats were randomly divided into control group(FiO_2=21%), hyperoxia group(FiO_2>85%) and hyperoxia + EPO group(FiO_2>85%). The newborn rats in the control group were placed in indoor air, and the rats in the hyperoxia and the hyperoxia+EPO group were placed in an indoor glass box with FiO_2 more than 85%. From the first day of the experiment, the rats in the hyperoxia + EPO group were intraperitoneally injected with EPO(800 U/kg), and the wet/dry mass(W/D) ratio of brain tissue was measured at day 1, 3, 7 and 14. Protein immunoblotting was used to detect the expression levels of CHOP and GRP78 protein in brain tissue. RESULTS The W/D values of brain tissue on the 1st, 3rd, 7th and 14th day of neonatal rats in the hyperoxia group were higher than those in the control group(P<0.05). The longer the exposure time under high oxygen, the more obvious the increase of W/D values. The water content of neonatal rat brain tissue in the hyperoxia + EPO group was obviously lower than that in the hyperoxia group(P<0.05). The expressions of CHOP and GRP78 protein in the hyperoxia group were significantly higher than those in the control group at day 1, 3, 7 and 14(P<0.05). The expression of CHOP and GRP78 protein in the hyperoxia + EPO group was significantly lower than those in the hyperoxia group(P<0.05). CONCLUSION EPO could alleviate cerebral edema induced by hyperoxia in neonatal rats and had neuroprotective effects on hyperoxic brain injury, which might be related to the down-regulation of CHOP and GRP78 protein expression.
引文
[1] Allin MP,Kontis D,Walshe M,et al..White matter and cognition in adults who were born preterm[J].Plos One,2011,6(10):e24525.
[2] Schrder M,Sutcliffe L.Consequences of stress in the secretory pathway:The ER stress response and its role in the metabolic syndrome[J].Methods Mol Biol,2010,648:43-62.
[3] Lozon TI,Eastman AJ,Matute-Bello G,et al..PKR-dependent CHOP induction limits hyperoxia-induced lung injury[J].Am J Physiol Lung Cell Mo Physiol,2011,300(3):422-429.
[4] 文建国,冯锦锦.促红细胞生成素的研究进展和临床应用[J].郑州大学学报:医学版,2015,50(6):733-736.
[5] Dericioglu N,Soylemezoglu F,Gursoy-zdemir Y,et al..Cell death and survival mechanisms are concomitantly active in the hippocampus of patients with mesial temporal sclerosis[J].Neuroscience,2013,237(6):56-65.
[6] 王永双,杨磊,许春花.重组人促红细胞生成素对高氧脑损伤新生大鼠IL-18,Caspase-1蛋白表达的影响[J].延边大学医学学报,2017,40(4):250-253.
[7] Du M,Tan Y,Liu G,et al..Effects of the Notch signalling pathway on hyperoxia-induced immature brain damage in newborn mice[J].Neuroscience Letters,2017,653:220-227.
[8] Liu Y,Jiang P,Du M,et al..Hyperoxia-induced immature brain injury through the TLR4 signaling pathway in newborn mice[J].Brain Res,2015,1610:51-60.
[9] Cao SS,Kaufman RJ.Endoplasmic reticulum stress and oxidative stress in cell fate decision and human disease[J].Antioxid Redox Signal,2014,21(3):396-413.
[10] Chen Y,Chang L,Li W,et al..Thioredoxin protects fetal type Ⅱ epithelial cells from hyperoxia-induced injury[J].Pediatr Pulmonol,2011,45(12):1192-1200.
[11] Feng J,Chen X,Sun X,et al..Expression of endoplasmic reticulum stress markers GRP78 and CHOP induced by oxidative stress in blue light-mediated damage of A2E-containing retinal pigment epithelium cells[J].Ophthalmic Res,2014,52(4):224-233.
[12] Li C,Harada A,Oh Y.IGFBP-3 sensitizes antiestrogen-resistant breast cancer cells through interaction with GRP78[J].Cancer Letters,2012,325(2):200-206.
[13] Sovolyova N,Healy S,Samali A,et al..Stressed to death-mechanisms of ER stress-induced cell death[J].Biol Chem,2014,395(1):1-13.
[14] Scull CM,Tabas I.Mechanisms of ER stress-induced apoptosis in atherosclerosis[J].Arterioscler Thromb Vasc Biol,2011,31(12):2792-2797.
[15] 高楚楚,卢红艳,唐炜.支气管肺发育不良大鼠肺GRP78及CHOP表达与肺细胞凋亡的关系[J].临床儿科杂志,2013,31(10):959-963.
[16] 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].中国新生儿科杂志,2016,31(3):206-211.
[17] Mastromarino V,Musumeci MB,Conti E,et al..Erythropoietin in cardiac disease:effective or harmful[J].J Cardiovascu Med,2013,14(12):870-878.
[18] Endesfelder S,Makki H,Haefen CV,et al..Neuroprotective effects of dexmedetomidine against hyperoxia-induced injury in the developing rat brain[J].Plos One,2017,12(2):e0171498.
[19] Zaghloul N,Nasim M,Patel H,et al..Overexpression of extracellular superoxide dismutase has a protective role against hyperoxia-induced brain injury in neonatal mice[J].Febs Journal,2012,279(5):871-881.
[2] Schrder M,Sutcliffe L.Consequences of stress in the secretory pathway:The ER stress response and its role in the metabolic syndrome[J].Methods Mol Biol,2010,648:43-62.
[3] Lozon TI,Eastman AJ,Matute-Bello G,et al..PKR-dependent CHOP induction limits hyperoxia-induced lung injury[J].Am J Physiol Lung Cell Mo Physiol,2011,300(3):422-429.
[4] 文建国,冯锦锦.促红细胞生成素的研究进展和临床应用[J].郑州大学学报:医学版,2015,50(6):733-736.
[5] Dericioglu N,Soylemezoglu F,Gursoy-zdemir Y,et al..Cell death and survival mechanisms are concomitantly active in the hippocampus of patients with mesial temporal sclerosis[J].Neuroscience,2013,237(6):56-65.
[6] 王永双,杨磊,许春花.重组人促红细胞生成素对高氧脑损伤新生大鼠IL-18,Caspase-1蛋白表达的影响[J].延边大学医学学报,2017,40(4):250-253.
[7] Du M,Tan Y,Liu G,et al..Effects of the Notch signalling pathway on hyperoxia-induced immature brain damage in newborn mice[J].Neuroscience Letters,2017,653:220-227.
[8] Liu Y,Jiang P,Du M,et al..Hyperoxia-induced immature brain injury through the TLR4 signaling pathway in newborn mice[J].Brain Res,2015,1610:51-60.
[9] Cao SS,Kaufman RJ.Endoplasmic reticulum stress and oxidative stress in cell fate decision and human disease[J].Antioxid Redox Signal,2014,21(3):396-413.
[10] Chen Y,Chang L,Li W,et al..Thioredoxin protects fetal type Ⅱ epithelial cells from hyperoxia-induced injury[J].Pediatr Pulmonol,2011,45(12):1192-1200.
[11] Feng J,Chen X,Sun X,et al..Expression of endoplasmic reticulum stress markers GRP78 and CHOP induced by oxidative stress in blue light-mediated damage of A2E-containing retinal pigment epithelium cells[J].Ophthalmic Res,2014,52(4):224-233.
[12] Li C,Harada A,Oh Y.IGFBP-3 sensitizes antiestrogen-resistant breast cancer cells through interaction with GRP78[J].Cancer Letters,2012,325(2):200-206.
[13] Sovolyova N,Healy S,Samali A,et al..Stressed to death-mechanisms of ER stress-induced cell death[J].Biol Chem,2014,395(1):1-13.
[14] Scull CM,Tabas I.Mechanisms of ER stress-induced apoptosis in atherosclerosis[J].Arterioscler Thromb Vasc Biol,2011,31(12):2792-2797.
[15] 高楚楚,卢红艳,唐炜.支气管肺发育不良大鼠肺GRP78及CHOP表达与肺细胞凋亡的关系[J].临床儿科杂志,2013,31(10):959-963.
[16] 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].中国新生儿科杂志,2016,31(3):206-211.
[17] Mastromarino V,Musumeci MB,Conti E,et al..Erythropoietin in cardiac disease:effective or harmful[J].J Cardiovascu Med,2013,14(12):870-878.
[18] Endesfelder S,Makki H,Haefen CV,et al..Neuroprotective effects of dexmedetomidine against hyperoxia-induced injury in the developing rat brain[J].Plos One,2017,12(2):e0171498.
[19] Zaghloul N,Nasim M,Patel H,et al..Overexpression of extracellular superoxide dismutase has a protective role against hyperoxia-induced brain injury in neonatal mice[J].Febs Journal,2012,279(5):871-881.