外源性神经生长因子和碱性成纤维细胞生长因子联合应用促进重型脑创伤模型大鼠内源性脑细胞的增殖
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摘要
背景:研究证明神经生长因子、碱性成纤维细胞生长因子能够明显在体外促进神经干细胞的自我更新及分化,二者联合应用对成年大鼠脑创伤后内源性脑细胞的影响研究甚少。目的:探索外源性神经生长因子和碱性成纤维细胞生长因子对重型脑创伤大鼠内源性大脑组织细胞的影响。方法:以改良Feeney氏法对48只SD大鼠进行脑创伤造模,造模后随机将48只大鼠分为神经生长因子组、碱性成纤维细胞生长因子组、联合组、对照组。造模后24 h,各组分别在脑室内注入对应的神经营养因子,对照组注射生理盐水。采用行为学实验观察肢体恢复情况,免疫组化法比较各组大鼠脑内BrdU阳性细胞的表达。结果与结论:①从造模第5天开始神经生长因子组、碱性成纤维细胞生长因子组、联合组行为学实验评分均小于对照组(P <0.05),且联合组评分低于神经生长因子组、碱性成纤维细胞生长因子组(P <0.05);②神经生长因子组、碱性成纤维细胞生长因子组、联合组BrdU阳性细胞数量明显多于对照组,差异有显著性意义(P <0.05),同时联合组BrdU阳性细胞明显多于神经生长因子组、碱性成纤维细胞生长因子组(P <0.05);③结果表明,外源性神经生长因子和碱性成纤维细胞生长因子可加快脑创伤大鼠肢体功能恢复和促进大脑组织细胞的增殖,且神经生长因子和碱性成纤维细胞生长因子的联合使用能取得更加显著的效果。
BACKGROUND: Either nerve growth factor or basic fibroblast growth factor can promote the self-renewal and differentiation of neural stem cells in vitro. Little has been reported on the effect of the combination of these two factors on endogenous brain cells after traumatic brain injury in adult rats.OBJECTIVE: To explore the effects of exogenous nerve growth factor and basic fibroblast growth factor on the proliferation of endogenous brain cells in the rats with severe traumatic brain injury.METHODS: Animal models of traumatic brain injury were made in 48 adult Sprague-Dawley rats using modified Feeney's method, and then randomized into 4 groups: nerve growth factor, basic fibroblast growth factor, combination(injection of nerve growth factor and basic fibroblast growth factor), and control(normal saline injection) groups. Intraventricular injection of different factors was performed correspondingly at 24 hours after modeling. The restoration of limb function was assessed through behavior observation. BrdU-positive cells in the brain were counted and compared among four groups using immunohistochemical method.RESULTS AND CONCLUSION:(1) Initially from the 5 th day after modeling, the behavioral test scores in the three treatment groups were significantly lower than those in the control group(P < 0.05), and the behavioral test scores in the combination group were significantly lower than those in the nerve growth factor and basic fibroblast growth factor groups(P < 0.05).(2) There were more BrdU-labeled cells in the three treatment groups than the control group(P < 0.05). The number of BrdU-positive cells in the combination group was significantly higher than that in the nerve growth factor and basic fibroblast growth factor groups(P < 0.05). In summary, the proliferation of brain cells and restoration of limb function after traumatic brain injury can be promoted by nerve growth factor or basic fibroblast growth factor. Morever, this effect can be considerably improved by the combination of nerve growth factor and basic fibroblast growth factor.
BACKGROUND: Either nerve growth factor or basic fibroblast growth factor can promote the self-renewal and differentiation of neural stem cells in vitro. Little has been reported on the effect of the combination of these two factors on endogenous brain cells after traumatic brain injury in adult rats.OBJECTIVE: To explore the effects of exogenous nerve growth factor and basic fibroblast growth factor on the proliferation of endogenous brain cells in the rats with severe traumatic brain injury.METHODS: Animal models of traumatic brain injury were made in 48 adult Sprague-Dawley rats using modified Feeney's method, and then randomized into 4 groups: nerve growth factor, basic fibroblast growth factor, combination(injection of nerve growth factor and basic fibroblast growth factor), and control(normal saline injection) groups. Intraventricular injection of different factors was performed correspondingly at 24 hours after modeling. The restoration of limb function was assessed through behavior observation. BrdU-positive cells in the brain were counted and compared among four groups using immunohistochemical method.RESULTS AND CONCLUSION:(1) Initially from the 5 th day after modeling, the behavioral test scores in the three treatment groups were significantly lower than those in the control group(P < 0.05), and the behavioral test scores in the combination group were significantly lower than those in the nerve growth factor and basic fibroblast growth factor groups(P < 0.05).(2) There were more BrdU-labeled cells in the three treatment groups than the control group(P < 0.05). The number of BrdU-positive cells in the combination group was significantly higher than that in the nerve growth factor and basic fibroblast growth factor groups(P < 0.05). In summary, the proliferation of brain cells and restoration of limb function after traumatic brain injury can be promoted by nerve growth factor or basic fibroblast growth factor. Morever, this effect can be considerably improved by the combination of nerve growth factor and basic fibroblast growth factor.
引文
[1]Chauhan NB.Chronic neurodegenerative consequences of traumatic brain injury.Restor Neurol Neurosci.2014;32(2):337-365.
[2]Wurzelmann M,Romeika J,Sun D.Therapeutic potential of brain-derived neurotrophic factor(BDNF)and a small molecular mimics of BDNF for traumatic brain injury.Neural Regen Res.2017;12(1):7-12.
[3]Gage FH.Mammalian neural stem cells.Science.2000;287(5457):1433-1438.
[4]Kernie SG,Parent JM.Forebrain neurogenesis after focal Ischemic and traumatic brain injury.Neurobiol Dis.2010;37(2):267-274.
[5]Li C,Che LH,Shi L,et al.Suppression of Basic Fibroblast Growth Factor Expression by Antisense Oligonucleotides Inhibits Neural Stem Cell Proliferation and Differentiation in Rat models With Focal Cerebral Infarction.J Cell Biochem.2017;118(11):3875-3882.
[6]Cui K,Zhou X,Luo J,et al.Dual gene transfer of bFGF and PDGF in a single plasmid for the treatment of myocardial infarction.Exp Ther Med.2014;7(3):691-696.
[7]Chen SQ,Cai Q,Shen YY,et al.Combined use of NGF/BDNF/bFGF promotes proliferation and differentiation of neural stem cells in vitro.Int J Dev Neurosci.2014;38:74-78.
[8]Feeney DM,Boyeson MG,Linn RT,Responses to cortical injury:I.Methodology and local effects of contusions in the rat.Brain Res.1981;211(1):67-77.
[9]刘阳,刘卫平,王孝安,等.神经生长因子与表皮生长因子干预创伤性脑损伤后内源性神经干细胞的增殖[J].中国组织工程研究,2012,16(1):65-69.
[10]Reynolds BA,Weiss S.Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system.Science.1992;255(5052):1707-1710.
[11]Yu TS,Washington PM,Kernie SG.Injury-Induced Neurogenesis:Mechanisms and Relevance.Neuroscientist.2016;22(1):61-71.
[12]Aleksandrova MA,Podgornyi OV,Marei MV,et al.Characteristics of human neural stem cells in vitro and after transplantation into rat brain.Bull Exp Biol Med.2005;139(1):114-120.
[13]Wang X,Gao X,Michalski S,et al.Traumatic Brain Injury Severity Affects Neurogenesis in Adult Mouse Hippocampus.J Neurotrauma.2016;33(8):721-733.
[14]Choi KC,Yoo DS,Cho KS,et al.Effect of single growth factor and growth factor combinations on differentiation of neural stem cells.J Korean Neurosurg Soc.2008;44(6):375-381.
[15]Lin Y,Wan JQ,Gao GY,et al.Direct hippocampal injection of pseudo lentivirus-delivered nerve growth factor gene rescues the damaged cognitive function after traumatic brain injury in the rat.Biomaterials.2015;69:148-157.
[16]Xiong LL,Chen ZW,Wang TH.Nerve growth factor promotes in vitro proliferation of neural stem cells from tree shrews.Neural Regen Res.2016;11(4):591-596.
[17]Chittka A.Differential regulation of SC1/PRDM4 and PRMT5mediated protein arginine methylation by the nerve growth factor and the epidermal growth factor in PC12 cells.Neurosci Lett.2013;550:87-92.
[18]Liu F,Xuan A,Chen Y,et al.Combined effect of nerve growth factor and brain?derived neurotrophic factor on neuronal differentiation of neural stem cells and the potential molecular mechanisms.Mol Med Rep.2014;10(4):1739-1745.
[19]Cirulli F,Alleva E.The NGF saga:from animal models of psychosocial stress to stress-related psychopathology.Front Neuroendocrinol.2009;30(3):379-395.
[20]Song JN,Liu ZW,Sui L,et al.Dynamic expression of nerve growth factor and its receptor TrkA after subarachnoid hemorrhage in rat brain.Neural Regen Res.2016;11(8):1278-1284.
[21]Mahmood A,Lu D,Wang L,et al.Intracerebral transplantation of marrow stromal cells cultured with neurotrophic factors promotes functional recovery in adult rats subjected to traumatic brain injury.J Neurotrauma.2002;19(12):1609-1617.
[22]Chiaretti A,Antonelli A,Genovese O,et al.Nerve growth factor and doublecortin expression correlates with improved outcome in children with severe traumatic brain injury.JTrauma.2008;65(1):80-85.
[23]Yeoh JS,de Haan G.Fibroblast growth factors as regulators of stem cell self-renewal and aging.Mech Ageing Dev.2007;128(1):17-24.
[24]Ramirez JJ,Finklestein SP,Keller J,et al.Basic fibroblast growth factor enhances axonal sprouting after cortical injury in rats.Neuroreport.1999;10(6):1201-1204.
[25]Caday CG,Klagsbrun M,Fanning PJ,et al.Fibroblast growth factor(FGF)levels in the developing rat brain.Brain Res Dev Brain Res.1990;52(1-2):241-246.
[26]Sun D,Bullock MR,McGinn MJ,et al.Basic fibroblast growth factor-enhanced neurogenesis contributes to cognitive recovery in rats following traumatic brain injury.Exp Neurol.2009;216(1):56-65.
[27]Zhang R,Zhang Z,Wang L,et al.Activated neural stem cells contribute to stroke-induced neurogenesis and neuroblast migration toward the infarct boundary in adult rats.J Cereb Blood Flow Metab.2004;24(4):441-448.
[28]Wang ZG,Cheng Y,Yu XC,et al.bFGF Protects Against Blood-Brain Barrier Damage Through Junction Protein Regulation via PI3K-Akt-Rac1 Pathway Following Traumatic Brain Injury.Mol Neurobiol.2016;53(10):7298-7311.
[2]Wurzelmann M,Romeika J,Sun D.Therapeutic potential of brain-derived neurotrophic factor(BDNF)and a small molecular mimics of BDNF for traumatic brain injury.Neural Regen Res.2017;12(1):7-12.
[3]Gage FH.Mammalian neural stem cells.Science.2000;287(5457):1433-1438.
[4]Kernie SG,Parent JM.Forebrain neurogenesis after focal Ischemic and traumatic brain injury.Neurobiol Dis.2010;37(2):267-274.
[5]Li C,Che LH,Shi L,et al.Suppression of Basic Fibroblast Growth Factor Expression by Antisense Oligonucleotides Inhibits Neural Stem Cell Proliferation and Differentiation in Rat models With Focal Cerebral Infarction.J Cell Biochem.2017;118(11):3875-3882.
[6]Cui K,Zhou X,Luo J,et al.Dual gene transfer of bFGF and PDGF in a single plasmid for the treatment of myocardial infarction.Exp Ther Med.2014;7(3):691-696.
[7]Chen SQ,Cai Q,Shen YY,et al.Combined use of NGF/BDNF/bFGF promotes proliferation and differentiation of neural stem cells in vitro.Int J Dev Neurosci.2014;38:74-78.
[8]Feeney DM,Boyeson MG,Linn RT,Responses to cortical injury:I.Methodology and local effects of contusions in the rat.Brain Res.1981;211(1):67-77.
[9]刘阳,刘卫平,王孝安,等.神经生长因子与表皮生长因子干预创伤性脑损伤后内源性神经干细胞的增殖[J].中国组织工程研究,2012,16(1):65-69.
[10]Reynolds BA,Weiss S.Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system.Science.1992;255(5052):1707-1710.
[11]Yu TS,Washington PM,Kernie SG.Injury-Induced Neurogenesis:Mechanisms and Relevance.Neuroscientist.2016;22(1):61-71.
[12]Aleksandrova MA,Podgornyi OV,Marei MV,et al.Characteristics of human neural stem cells in vitro and after transplantation into rat brain.Bull Exp Biol Med.2005;139(1):114-120.
[13]Wang X,Gao X,Michalski S,et al.Traumatic Brain Injury Severity Affects Neurogenesis in Adult Mouse Hippocampus.J Neurotrauma.2016;33(8):721-733.
[14]Choi KC,Yoo DS,Cho KS,et al.Effect of single growth factor and growth factor combinations on differentiation of neural stem cells.J Korean Neurosurg Soc.2008;44(6):375-381.
[15]Lin Y,Wan JQ,Gao GY,et al.Direct hippocampal injection of pseudo lentivirus-delivered nerve growth factor gene rescues the damaged cognitive function after traumatic brain injury in the rat.Biomaterials.2015;69:148-157.
[16]Xiong LL,Chen ZW,Wang TH.Nerve growth factor promotes in vitro proliferation of neural stem cells from tree shrews.Neural Regen Res.2016;11(4):591-596.
[17]Chittka A.Differential regulation of SC1/PRDM4 and PRMT5mediated protein arginine methylation by the nerve growth factor and the epidermal growth factor in PC12 cells.Neurosci Lett.2013;550:87-92.
[18]Liu F,Xuan A,Chen Y,et al.Combined effect of nerve growth factor and brain?derived neurotrophic factor on neuronal differentiation of neural stem cells and the potential molecular mechanisms.Mol Med Rep.2014;10(4):1739-1745.
[19]Cirulli F,Alleva E.The NGF saga:from animal models of psychosocial stress to stress-related psychopathology.Front Neuroendocrinol.2009;30(3):379-395.
[20]Song JN,Liu ZW,Sui L,et al.Dynamic expression of nerve growth factor and its receptor TrkA after subarachnoid hemorrhage in rat brain.Neural Regen Res.2016;11(8):1278-1284.
[21]Mahmood A,Lu D,Wang L,et al.Intracerebral transplantation of marrow stromal cells cultured with neurotrophic factors promotes functional recovery in adult rats subjected to traumatic brain injury.J Neurotrauma.2002;19(12):1609-1617.
[22]Chiaretti A,Antonelli A,Genovese O,et al.Nerve growth factor and doublecortin expression correlates with improved outcome in children with severe traumatic brain injury.JTrauma.2008;65(1):80-85.
[23]Yeoh JS,de Haan G.Fibroblast growth factors as regulators of stem cell self-renewal and aging.Mech Ageing Dev.2007;128(1):17-24.
[24]Ramirez JJ,Finklestein SP,Keller J,et al.Basic fibroblast growth factor enhances axonal sprouting after cortical injury in rats.Neuroreport.1999;10(6):1201-1204.
[25]Caday CG,Klagsbrun M,Fanning PJ,et al.Fibroblast growth factor(FGF)levels in the developing rat brain.Brain Res Dev Brain Res.1990;52(1-2):241-246.
[26]Sun D,Bullock MR,McGinn MJ,et al.Basic fibroblast growth factor-enhanced neurogenesis contributes to cognitive recovery in rats following traumatic brain injury.Exp Neurol.2009;216(1):56-65.
[27]Zhang R,Zhang Z,Wang L,et al.Activated neural stem cells contribute to stroke-induced neurogenesis and neuroblast migration toward the infarct boundary in adult rats.J Cereb Blood Flow Metab.2004;24(4):441-448.
[28]Wang ZG,Cheng Y,Yu XC,et al.bFGF Protects Against Blood-Brain Barrier Damage Through Junction Protein Regulation via PI3K-Akt-Rac1 Pathway Following Traumatic Brain Injury.Mol Neurobiol.2016;53(10):7298-7311.
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