胞红蛋白在缺氧神经元中的表达

doi:10.11852/zgetbjzz2014-22-05-14

中国儿童保健杂志 ›› 2014, Vol. 22 ›› Issue (5): 492-496.DOI: 10.11852/zgetbjzz2014-22-05-14

胞红蛋白在缺氧神经元中的表达

马海然,史雪川

汕头大学医学院第二附属医院儿科,广东汕头 515041

收稿日期:2013-11-13 发布日期:2014-05-10 出版日期:2014-05-10
通讯作者: 史雪川, E-mail:shixuechuan@163.com
作者简介:马海然,男,硕士学位,研究方向为新生儿缺氧缺血性脑病方面。
基金资助:
广东省科技计划项目(2008B080701036);广东省医学科研基金立项(B2009180)

Expression of cytoglobin in hypoxic neurons in vitro

MA Hai-ran,SHI Xue-chuan

Department of Pediatrics,the Second Affiliated Hospital,Shantou Medical College,Shantou,Guangdong 515041,China

Received:2013-11-13 Online:2014-05-10 Published:2014-05-10
Contact: SHI Xue-chuan,E-mail:shixuechuan@163.com

摘要/Abstract

摘要： 目的探讨细胞红蛋白(cytoglobin,CYGB)在体外培养神经元缺氧时的表达,为研究缺氧缺血性脑病(hypoxic ischemic encephalopathy,HIE)的生理病理机制及寻求该病新的治疗方法提供实验基础。方法运用无血清培养技术进行胎鼠皮质神经元原代培养6 d;先用神经元特异性烯醇化酶免疫组织化学方法纯度鉴定,然后随机分配为4组,将其中3组(第1组作为对照)置于0.5%~0.9%的O₂中,用CCK8试剂盒测试细胞生存率,同时用Real-timePCR和Westernblot观察CYGB在神经元正常及缺氧不同时间点(8 h,16 h,24 h)核酸及蛋白的表达情况。结果神经元纯度鉴定纯度大于99%;缺氧后神经元生存率随着时间推移降低,但绝大部分神经元仍存活;CYGB核酸及蛋白的表达随缺氧时间增长而递增,且每组之间差异有统计学意义(P<0.05)。结论在体外培养的神经元细胞中,缺氧可以促进CYGB的表达,提示CYGB在缺氧性神经损伤中可能发挥某种特定的生理功能。

关键词: 细胞红蛋白, 原代神经元, 缺氧缺血性脑损伤

Abstract: Objective To examine the expression of cytoglobin(CYGB) of neurons in vitro when suffered hypoxia,which provides an experimental basis for exploring the physiological and pathological mechanism and seeking new treatments of hypoxic-ischemic encephalopathy. Methods The primary cortical neurons from embryonic rat were cultured in a serum-free culture system of B27-supplemented neurobasal medium.The neurons were identified by polyclonal antibody against neuron specific enolase(NSE) on the 6th day.They were divided into 4 groups randomly for assessing cell viability by CCK8 and observing the expression of CYGB by Real-time PCR and Westernblot at 0、8、16、24 h after hypoxic model(oxygen concentration between 0.5% and 0.9%) was induced. Results The purity of neurons was greater than 99%;and cell viability were slightly reduced after hypoxia.Expression of RNA and protein of CYGB was increased in a time-dependent manner.And there was statistical differenc in each group (P<0.05). Conclusions CYGB can up-regulate in neurons in vitro after hypoxia,indicating CYGB may play certain important roles in neurons under hypoxia.

Key words: cytoglobin, neurons, hypoxic-ischemic encephalopathy

中图分类号:

Q95-33

马海然,史雪川. 胞红蛋白在缺氧神经元中的表达[J]. 中国儿童保健杂志, 2014, 22(5): 492-496.

MA Hai-ran,SHI Xue-chuan. Expression of cytoglobin in hypoxic neurons in vitro[J]. journal1, 2014, 22(5): 492-496.

参考文献

[1] Kawada N,Kristensen DB,Asahina K,et al.Characterization of a stellate cell activation-associated protein (STAP) with peroxidase activity found in rat hepatic stellate cells[J].Journal of Biological Chemistry,2001,276(27):25318-25323.
[2] Sugimoto H,Makino M,Sawai H,et al.Structural basis of human cytoglobin for ligand binding[J].Journal of Molecular Biology,2004,339(4):873-885.
[3] Trent JT,Hargrove MS.A ubiquitously expressed human hexacoordinate hemoglobin[J].The Journal of Biological Chemistry,2002,277(22):19538-19545.
[4] Burmester T,Ebner B,Weich B,et al.Cytoglobin:a novel globin type ubiquitously expressed invertebrate tissues[J].Molecular Biology and Evolution,2002,19(4):416-421.
[5] Geuens E,Brouns I,Flamez D,et al.A globin in the nucleus![J].The Journal of Biological Chemistry,2003,278(33):30417-30420.
[6] Hundahl CA,Allen GC,Hannibal J,et al.Anatomical characterization of cytoglobin and neuroglobin mRNA and protein expression in the mouse brain[J].Brain research,2010,1331:58-73.
[7] Mammen PP,Shelton JM,Ye Q,et al.Cytoglobin is a stress-responsive hemoprotein expressed in the developing and adult brain[J].The Journal of Histochemistry and Cytochemistry:Official Journal of the Histochemistry Society,2006,54(12):1349-1361.
[8] Nakatani K,Okuyama H,Shimahara Y,et al.Cytoglobin/STAP,its unique localization in splanchnic fibroblast-like cells and function in organ fibrogenesis[J].Laboratory investigation,2003,84(1):91-101.
[9] Schmidt M,Gerlach F,Avivi A,et al.Cytoglobin is a respiratory protein in connective tissue and neurons,which is up-regulated by hypoxia[J].The Journal of biological chemistry,2004,279(9):8063-8069.
[10] Shigematsu A,Adachi Y,Matsubara J,et al.Analyses of expression of cytoglobin by immunohistochemical studies in human tissues[J].Hemoglobin,2008,32(3):287-296.
[11] Man KN,Philipsen S,Tan-Un KC.Localization and expression pattern of cytoglobin in carbon tetrachloride-induced liver fibrosis[J].Toxicology letters,2008,183(1-3):36-44.
[12] Li RC,Lee SK,Pouranfar F,et al.Hypoxia differentially regulates the expression of neuroglobin and cytoglobin in rat brain[J].Brain research,2006,1096(1):173-9.
[13] Ostojic J,Sakaguchi DS,de Lathouder Y,et al.Neuroglobin and cytoglobin:oxygen-binding proteins in retinal neurons[J].Investigative ophthalmology & visual science,2006,47(3):1016-1023.
[14] Fordel E,Geuens E,Dewilde S,et al.Cytoglobin expression is upregulated in all tissues upon hypoxia:an in vitro and in vivo study by quantitative real-time PCR[J].Biochemical and Biophysical Research Communications,2004,319(2):342-348.
[15] Tian SF,Yang HH,Xiao DP,et al.Mechanisms of neuroprotection from hypoxia-ischemia (HI) brain injury by up-regulation of cytoglobin (CYGB) in a neonatal rat model[J].The Journal of Biological Chemistry,2013,288(22):15988-16003.
[16] Erecińska M,Silver IA.Tissue oxygen tension and brain sensitivity to hypoxia[J].Respiration Physiology,2001,128(3):263-276.
[17] Guo X,Philipsen S,Tan-Un KC.Study of the hypoxia-dependent regulation of human CYGB gene[J].Biochemical and Biophysical Research Communications,2007,364(1):145-150.
[18] Burmester T,Gerlach F,Hankeln T.Regulation and role of neuroglobin and cytoglobin under hypoxia[J].Advances in experimental medicine and biology,2007,618:169-180.
[19] Hamdane D,Kiger L,Dewilde S,et al.The redox state of the cell regulates the ligand binding affinity of human neuroglobin and cytoglobin[J].Journal of Biological Chemistry,2003,278(51):51713-51721.
[20] Trandafir F,Hoogewijs D,Altieri F,et al.Neuroglobin and cytoglobin as potential enzyme or substrate[J].Gene,2007,398(1-2):103-113.
[21] Petersen MG,Dewilde S,Fago A.Reactions of ferrous neuroglobin and cytoglobin with nitrite under anaerobic conditions[J].Journal of inorganic biochemistry,2008,102(9):1777-1782.
[22] Hodges NJ,Innocent N,Dhanda S,et al.Cellular protection from oxidative DNA damage by over-expression of the novel globin cytoglobin in vitro[J].Mutagenesis,2008,23(4):293-298.
[23] Johnston MV,Trescher WH,Ishida A,et al.Neurobiology of hypoxic-ischemic injury in the developing brain[J].Pediatric research,2001,49(6):735-741.
[24] Aliev G,Smith MA,Obrenovich ME,et al.Role of vascular hypoperfusion-induced oxidative stress and mitochondria failure in the pathogenesis of Alzheimer disease[J].Neurotoxicity research,2003,5(7):491-504.
[25] Li D,Chen XQ,Li WJ,et al.Cytoglobin up-regulated by hydrogen peroxide plays a protective role in oxidative stress[J].Neurochemical Research,2007,32(8):1375-1380.

胞红蛋白在缺氧神经元中的表达

Expression of cytoglobin in hypoxic neurons in vitro

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