摘要
目的探讨急性脊髓损伤后热休克蛋白70(heat shock prtein 70,HSP70)表达的变化规律;通过诱导急性脊髓损伤后早期HSP70的大量表达,探讨其诱导剂替普瑞酮(Geranylgeranylacetone,GGA)修复脊髓损伤的疗效,综合、客观评价脊髓功能恢复程度,从而寻求一种简单有效、可应用于临床的新方法,为临床治疗脊髓损伤提供实验学依据。
方法1、急性脊髓损伤后HSP70表达的变化。健康成年雌性Wistar大鼠50只,随机分为Ⅰ假手术组:单纯显露T10节段脊髓,不予打击损伤;Ⅱ损伤组:显露T10节段脊髓,使用MASCIS Impactor ModelⅡ脊髓损伤打击器打击脊髓(10g×25mm),建立急性脊髓损伤模型。分别于术后6、12、24、48、72小时,每组随机选取5只动物,取材、固定、切片,行HSP70免疫组织化学染色,观察HSP70表达变化情况。
2、GGA对急性脊髓损伤后HSP70表达、神经细胞凋亡及神经功能恢复的影响。健康成年雌性Wistar大鼠75只,随机分为3组,各25只。Ⅰ对照组:损伤前24小时、损伤后半小时以及损伤后的每24小时至伤后1周分别经胃管给予等量生理盐水;Ⅱ损伤后给药组:损伤前24小时经胃管给予等量生理盐水,损伤后半小时以及损伤后的每24小时至伤后1周均经胃管给予GGA(溶于生理盐水)600mg/kg;Ⅲ损伤前、后联合给药组:各时间点分别经胃管给予GGA(溶于生理盐水)600mg/kg。所有动物均手术显露T10节段脊髓,使用MASCISImpactor ModelⅡ脊髓损伤打击器打击脊髓(10g×25mm),建立急性脊髓损伤模型。免疫组织化学染色测定HSP70的表达情况,TUNEL法检测神经细胞凋亡情况,HE染色光学显微镜观察脊髓组织形态学改变,BBB评分法进行行为学功能评价。
结果1、急性脊髓损伤后HSP70表达水平迅速上升,至伤后24h达到高峰,之后转为下降,至伤后72h脊髓组织内已少见HSP70阳性细胞;而假手术组仅有低水平、短时间的HSP70表达。
2、各组术后12、24、48、72小时HSP70表达水平差异有统计学意义(ANOVA,P<0.05),Ⅲ组、Ⅱ组均大于Ⅰ组。各组术后12、24、48、72小时损伤部位细胞凋亡水平差异有统计学意义(ANOVA,P<0.05),Ⅲ组、Ⅱ组均小于Ⅰ组。所有大鼠在脊髓损伤后出现双下肢截瘫,BBB评分组间比较,损伤后7天至伤后28天,各组间差异有统计学意义(ANOVA,P<0.05),Ⅲ组、Ⅱ组评分大于Ⅰ组。
结论急性脊髓损伤后HSP70表达自发性上调,但其上调的水平有限,持续时间较短。GGA可以增加急性脊髓损伤后HSP70的表达水平,减少急性脊髓损伤后神经细胞的凋亡,促进急性脊髓损伤后神经功能的恢复。
Objective To observe the expression of heat shock prtein 70(HSP70) after acute spinal cord injury(ASCI);and also observe the effect of a HSP70 inducer -Geranylgeranylacetone(GGA) on repairing of ASCI through up-regulation HSP70 expression in the earlier period after ASCI.Evaluation the functional recovery objectively in many ways to find an new effective way,which can be used in clinical practice easily,for providing an experimental evidence to clinical treatment of ASCI.
Methods 1.The expression of HSP70 after acute spinal cord injury.50 healthy female Wistar rats were separated into two groups randomly(25 rats in each group ):Ⅰcontrol group,rats were exposed T10 spinal cord,but not injured by the impactor;Ⅱstudy group,rats were exposed T10 spinal cord,which were then injured by the impactor(10g×25mm).Rats were sacrificed 6,12,24,48,72 hours after the operation separativly,and the injured spinal cord were taken out for fix,slice,and Immunohistochemistry staining was used to determin HSP70 expression.
2.The effect of GGA on expression of HSP70,apoptosis of neural cells and functional recovery after ASCI.75 healthy female Wistar rats were separated into three groups randomly(25 rats in each group):Ⅰcontrol group,Ⅱpost injury GGA administration group andⅢahead and post injury combined GGA administration group.Rats in groupⅠwere administrated equal saline by oral tube 24h ahead of injury,0.5h after injury and every 24h after injury until 1 week later;rats in groupⅡwere administrated equal saline 24h ahead of injury and GGA(600 mg/kg) 0.5h after injury and every 24h after injury until 1 week later;rats in groupⅢwere administrated GGA(600 mg/kg) on all the time points.Immunohistochemistry staining was used to determin HSP70 expression,TUNEL staining was used to detect the apoptosis level,HE staining and optics microscope was used to observe the histomorphology change of the spinal cord and the BBB(Basso,Beattie and Bresnahan) scales were used to evaluate hindlimb neurological function.
Results 1.The expression level of HSP70 were upgraded rapidly after ASCI,the peak value were presented at 24h after injury,then the value decreased,and to be rare 72h after injury.In the control group,the expression of HSP70 was in a lower lever, and for a shorter period.
2.The expression level of HSP70 among groups were statistically significance 12、24、48、72h after injury(ANOVA,P<0.05),groupⅢandⅡexceed groupⅠ; the apoptosis level among groups were statistically significance 12、24、48、72h after injury(ANOVA,P<0.05) too,groupⅢandⅡwere lower than groupⅠ.All the rats were inferior paraplegia after injury.The BBB score among groups were statistically significance since 7 days after injury(ANOVA,P<0.05),groupⅢandⅡexceed groupⅠ.
Conclusions The expression level of HSP70 was upgraded spontaneously.But the degree of it was limited,and the period of upregulation is deficient.GGA can increase the level of HSP70 expression,reduce the apoptosis of neural cells and promote the functional recovery after acute spinal cord injury.
引文
[1]Shi R,Pryor JD.Pathological changes of isolated spinal cord axons in response to mechanical stretch[J].Neuroscience,2002;110(4):765-777.
[2]Xu P,Gong WM,Li Y,et al.Destructive pathological changes in the rat spinal cord due to chronic mechanical compression[J].Neurosurg Spine,2008;8(3):279-285.
[3]Walmsley AR,Mir AK.Targeting the Nogo-A euritis pathway to promote recovery following acute CNS injury[J].Curr Pharm Des,2007;13(24):2470-2484.
[4]Adams M,Carlstedt T,Cavanagh J,et al.International spinal research trust research strategy:a discussion document[J].Spinal Cord,2007;45(1):2-14.
[5]Feng SQ,Kong XH,Guo SF,et al.Treatment of Spinal Cord Injury with Co-Grafts of Genetically Modified Schwann Cells and Fetal Spinal Cord Cell Suspension in the Rat[J].Neurotoxicity Research,2005;7(1,2):169-177.
[6]Lee KH,Suh-Kim H,Choi JS,et al.Human mesenchymal stem cell transplantation promotes functional recovery following acute spinal cord injury in rats[J].Acta Neurobiol Exp,2007;67(1):13-22.
[7]Garbossa D,Fontanella M,Fronda C,et al.New strategies for repairing the injured spinal cord:the role of stem cells[J].Neurol Res,2006;28(5):500-504.
[8]冯世庆,周先虎,孔晓红,等.自体激活雪旺细胞移植治疗急性脊髓损伤的实验研究[J].中华骨科杂志,2006;26(8):553-557.
[9]Oudega M,Moon LD,de Almeida Leme RJ.Schwann cells for spinal cord repair[J].Braz J Med Biol Res.2005;38(6):825-35.
[10]Feron F,Perry C,Cochrane J,et al.Autologous olfactory ensheathing cell transplantation in human spinal cord injury[J].Brain,2005;128(12):2951-2960.
[11]冯世庆.雪旺细胞移植治疗脊髓损伤研究进展[J].中国现代神经疾病杂志2004;24:276-279.
[12]Feng SQ,Kong XH,Guo SF,et al.Use of engineered peripheral nerve autografts for spinal cord repair[J].Neuroreport,2006;12:102-110. 与临床研究初步报告[C].中华医学会第七届全国骨科学术会议汇编,广州,2004.302.
[14]Fujiki M,Kobayashi H,Abe T.Astroglial activation accompanies heat shock protein upregulation in rat brain following single oral dose of Geranylgeranylacetone[J].Brain Res,2003;991:254-257.
[15]Uchida S,Fujiki M,Nagai Y,et al.Geranylgeranylacetone,a noninvasive heat shock protein inducer,induces protein kinase C and lead to neuroprotection against cerebral infarction in rats[J].Neuroscience Letters,2006;396:220-224.
[16]Yasuda H,Shichinohe H,Kuroda S.Neuroprotective effect of a heat shock protein inducer geranylgeranylacetone in permanent focal cerebral ischemic.Brain Research[J].2005;1032:176-182.
[17]Nagai Y,Fujiki M,Inove R.Neuroprotective effect of geranylgeranylacetone,a noninvasive heat shock protein inducer on cerebral infarction in rats[J].Neuroscience Letters,2005;374:183-188.
[18]Yamanaka K,Takahashi N,Ooie T,et al.Role of protein kinase C in geranylgeranylacetone induced expression of heat shock protein 72 and Cardioprotection in the rat heart[J].Journal of molecular and Cellular Cardiology,2003;35:785-794.
[19]Sato K,Saito H,Matsuki N,et al.HSP70 is essential to the neuroprotective effect of heat shock[J].Brain Res,1996;740:117-123.
[20]Hopkins DA,Plumier J,Currie RW.Induction of the 27-kDa heat shock protein (Hsp27) in the rat medulla oblongata after vagus nerve injury[J].Experimental Neurology,1998;153:173-183
[21]Zarah B,Josephine N,Heather D.Nonsteroidal anti-inflammatory drugs differentially affect the heat shock response in cultured spinal cord cells[J].Cell Stress &Chaperones,2005;10(3),185-196.
[22]Valen Guro,Tsutomu Kawakami,Peeter Tohepold,et al.Glucocorticoid pretreatment protects cardiac function and induces cardiac heat shock protein 72[J].Am J Physiol Heart Circ Physiol,2000;279:H836-H843.
[23]Brown IR.Heat shock proteins and protection of the nervous system[J].Ann N Y Acad Sci,2007;1113:147-158.
[24]Morimoto R.Cell in stress:transcriptional activation of heat shock genes[J].Science,1993;259:1409-1410.
[25]Zarah B,Josephine N,Heather D.Nonsteroidal anti-inflammatory drugs differentially affect the heat shock response in cultured spinal cord cells[J].Cell Stress &Chaperones,2005;10(3),185-196.
[26]Basso DM,Beattie MS,Bresnahan JC.A sensitive and reliable locomotor rating scale for open field testing in rats[J].J Neurotrauma,1995;12(1):1-21.
[27]Growe MJ,Bresnahan JC,Shuman SL,et al.Apoptosis and delayed degeneration after spinal cord injury in rats and monkey[J].Nature Medicine,1997;3:73-76.
[28]Liu XZ,Xu XM,Hu R,et al.Neuronal and glial apoptsis after traumatic spinal cord injury[J].J Neurosci,1997;17:5394-5406.
[29]Yong C,Arnold PM,Zoubine MN,et al.Apoptosis cellular compartments of rats spinal cord after severe contusion injury[J].J Neurotrauma,1998;15:459-472.
[30]Whalley K,O Neill P,Ferretti P.Changes in response to spinal cord injury with development:vascularization,hemorrhage and apoptosis[J].Neuroscience,2006;137:821-832.
[31]Casha S,Yu W R,Fehlings MG.FAS deficiency reduces apoptosis,spares axons and improves function after spinal cord injury[J].Exp Neurol,2005;196:390-400.
[32]邵将,贾连顺,曹师锋等.热休克蛋白70在大鼠急性打击损伤脊髓中的表达[J].脊柱外科杂志,2008;6(3):169-172.
[33]Zhang P,Abraham VS,Kraft KR.Hyperthermic preconditioning protects against spinal cord ischemic injury[J].Annals of Thoracic Surgery,2001;70:1490-1495.
[34]王勇,张永福,毛伯镛.热休克预处理在大鼠脊髓损伤后细胞凋亡中的作用[J].医药论坛杂志,2005;26(13):43-45.
[35]Motoyoshi N,Sakurai M,Hayashi T,et al.Establishment of a local cooling model against spinal cord ischemia representing prolonged induction of heat shock protein[J].Thorac&Cardiovasc Surg,2001;122:351-357.
[36]Perdrizet,George A,Lena.Preoperative stress conditioning prevents paralysis after experimental aortic surgery:Increased heat shock protein content is associated with ischemic tolerance of the spinal cord[J].Thoracic Cardiovascular Surgery,2002;124:162-170.
[37]De Bosscher,K Vanden,Berghe W H.Mechanisms of anti-inflammatory action and of immunosuppression by glucocorticoids:negative interference of activated glucocorticoid receptor with transcription factors[J].J Neuroimmunol,2000;109:16-22.
[38]Kalmar B,Burnstock G.The efect of neonatal nerve injury on the expression of heat shock proteins in developing rat motoneurones[J].Neurotrauma,2002;19:667-679.
[39]管华清,杨惠林,徐浩等.脊髓缺血再灌注损伤后神经元凋亡与热休克蛋白:部分还是全部影响[J].中国临床康复杂志,2005;9(25):47-49.
[40]Parcellier A,Gurbuxani S.Heat shock proteins,cellular chaperones that modulate mitochondrial cell death pathways[J].Biophysical Research Communications,2003;304(3):505-512.
[41]Liu WL,Chen SJ,Chen Y,et al.Protective effects of heat shock protein70induced by geranylgeranylacetone in atrophic gastritis in rats[J].Acta Pharmacol Sin,2007;28(7):1001-1006.
[42]Sinn DI,Chu K and Lee ST,et al.Pharmacological induction of heat shock protein exerts neuroprotective effects in experimental intracerebral hemorrhage[J].Brain Res,2007,1135(1):167-176.
[43]Fujiki M,Furukawa Y,Kobayashi H,et al.Geranylgeranylacetone limits secondary injury,neuronal death,and progressive necrosis and cavitation after spinal cord injury.2005;1053:175-184.
[44]Mulligan MS,Varani j,Dame MK,et al.Role of endothelial-leukocyte adhesion molecule-1(ELAM-1) in neutrophil-mediated lung injury in rats[J].Clin Invest.1991;88:1396-1406.
[45]Taoka K,Okajima K.Spinal cord injury in the rat[J].Prog.Neurobiol.1998,56:341-358.
[46]Taoka K,Okajima K,Uchiba M,et al.Activated protein C reduces the severity of compression-induced spinal cord injury in rats by inhibiting activation of leukocytes[J].Neurosci,1998;18:1393-1398.
[47]Wojcik C,Di Naopoli,N.Ubiquitin-proteasome system and proteasome inhibition:new strategies in stroke therapy[J].Stroke,2004;35,1506-1518.
[48]Yenari MA,Liu J,Zheng,Z,et al.Antiapoptotic and anti-inflammatory mechanisms of heat-shock protein protection[.T].Ann.N.Y.Acad.Sci.2005;1053,74-83.
[49]Hirakawa T,Rokutan K,Nikawa T,et al.Geranylgeranylacetone induces heat shock proteins in cultured guinea pig gastric mucosal cells and rat gastric mucosa[J].Gastroenterology,1996;111:345-357.
[50]Masuda Y,Sumita S,Fujimura N,et al.Geranylgeranylacetone attenuates septic diaphragm dysfunction by induction of heat shock protein 70[J].Crit Care Med,2003;31:2585-2591.
[51]Mizushima T,Tsutsumi S,Rokutan K,et al.Suppression of ethanol-induced apoptotic DNA fragmentation by geranylgeranylacetone in cultured guinea pig gastric mucosal cell[J]s.Dig.Dis.Sci.,1999;44:510-514.
[52]Ooie T,Takahashi N,Saikawa T,et al.Single oral dose of geranylgeranylacetone induces heat-shock protein 72 and renders protection against ischemia/ reperfusion injury in rat heart[J].Circulation,2001;104:1837-1843.
[53]Yamanaka K,Takahashi N,Ooie T,et al.Role of protein kinase C in geranylgeranylacetone-induced expression of heat-shock protein 72 and cardioprotection in the rat heart[J].Mol.Cell.Cardiol,2003;35:785-794.
[54]Bar-Or D,Proposed mechanism for induction of heat shock protein 70 by geranylgeranyl acetone by prenylation of Ras proteins[J].Crit.Care Med.2003;31:2700-2701.
[55]Dong IS,Kon C,Soon TL,et al.Pharmacological induction of heat shock protein exerts neuroprotective effects in experimental intracerebral hemorrhage[J].Brain research,2007;1135:167-176.
[56]Kastuno M.,Sang C,Adachi H.,et al.Pharmacological induction of heat-shock proteins alleviates polyglutamine-mediated motor neuron disease[J].Proc.Natl Acad.Sci.,2005;102:16801-16806.
[57]Luo H.R.,Hattori H.,Hossain M.A.,et al.Akt as mediator of cell death[J].Proc.Natl Acad.Sci.,2003;100:11712-11717.
[58]Zou Y.,Zhu W.,Sakamoto M.,et al.Heat shock transcription factor 1 protects cardiomyocytes from ischemia/reperfusion injury[J].Circulation,2003;108:3024-3030.
[59]Kim H.S.,Skurk C,Maatz H.,et al.Akt/FOXO3a signaling modulates the endothelial stress response through regulation of heat shock protein 70 expression^.Faseb J,2005;19:1042-1044.
[60]Kretz A,Happold CJ,Marticke JK,et al.Erythropoietin promotes regeneration of adult CNS neurons via Jak2/Stat3 and PI3K/AKT pathway activation[J].Mol.Cell.Neurosci,2005;29:569-579.
[61]Stephanou A.,Latchman DD.Transcriptional regulation of the heat shock protein genes by STAT family transcription factors[J].Gene Expression,1999;7:311-319.
[62]D Liu,G.Y Xu,E.Pan,et al.Neurotoxicity of glutamate at the concentration released upon spinal cord injury[.T].Neuroscience,1999;93:1383-1389.
[63]T.Mizushima,S.Tsutsumi,K.Rokutan,et al.Suppression of ethanol-induced apoptotic DNA fragmentation by geranylgeranylacetone in cultured guinea pig gastric mucosal cells[J].Dig.Dis.Sci.,1999;44:510-514.
[64]Yasuda H,Shichinohe H,Kuroda S,et al.Neuroprotective effect of a heat shock protein inducer,geranylgeranylacetone in permanent focal cerebral ischemia[J].Brain Research,2005;1032:176-182.
[65]Fujiki M.,Kobayashi H.,Inoue R.,et al.Single oral dose of geranylgeranylacetone for protection against delayed neuronal death induced by transient ischemia[J].Brain Res.2004a;1020:210-213.
[66]Fujiki M.,Kobayashi H.,Inoue R.,et al.A single oral dose of geranylgeranylacetone attenuates kainic acid-induced seizure and neuronal cell death in rat hippocampus[J].Brain Res,2004b;1021:281-285.
[67]Sumigawa S.,Nakao T.,Iwama T,et al.Acute toxicity test of tetraprenylacetone in rats and mice[J].Prog.Med.1983;3:1053-1057.
[68]Rivlin AS,Tator CH.Objective clinical assessment of motor function after experimental spinal cord injury in the rat[J].J Neurosurg,1977;47(4):577-581.
[69]Gale K,Kerasidis H,Wrathall JR.Spinal cord contusion in the rat:behavioral analysis of functional neurologic impairment[J].Exp Neurol,1985;88(1):123-134.
[1]Ackery A,Tator C,Krassioukov A.A global perspective on spinal cord injury epidemiology.J Neurotrauma,2004;21(10):1355-1370.
[2]Sato K,Saito H,Matsuki N,et al.HSP70 is essential to the neuroprotective effect of heat shock[J].Brain Res,1996;740:117-123.
[3]Allen GV,Gerami D,Esser MJ.Conditioning effects of repetitive mild neurotrauma on motor function in an animal model of focal brain injury[J].Neuroscience,2000;99:93-105.
[4]Wald K,Ito M,Miyazawa T,et al.Repeated hyperbaric oxygen induces ischemic tolerance in gerbil hippocampus[J].Brain Res,1996;740:15-20.
[5]Brown IR.Heat shock proteins and protection of the nervous system[J].Ann N Y Acad Sci,2007;1113:147-158.
[6]Franklin T B,Krueger-naug A M,Clarke D B,et al.The role of heat shock proteins HSP70 and Hsp27 in cellular protection of the central nervous system[J].Int J Hyperthermia,2005;21(5):379-392.
[7]Morimoto R.Cell in stress:transcriptional activation of heat shock genes[J].Science,1993;259:1409-1410.
[8]Kalmar B,Burnstock G.The efect of neonatal nerve injury on the expression of heat shock proteins in developing rat motoneurones[J].Neurotrauma,2002;19:667-679.
[9]Zhang P,Abraham VS,Kraft KR.Hyperthermic preconditioning protects against spinal cord ischemic injury[J].Annals of Thoracic Surgery,2001;70:1490-1495.
[10]王勇,张永福,毛伯镛.热休克预处理在大鼠脊髓损伤后细胞凋亡中的作用[J].医药论坛杂志,2005;26(13):43-45.
[11]Motoyoshi N,Sakurai M,Hayashi T,et al.Establishment of a local cooling model against spinal cord ischemia representing prolonged induction of heat shock protein[J].Thorac&Cardiovasc Surg,2001;122:351-357.
[12]Perdrizet,George A,Lena.Preoperative stress conditioning prevents paralysis after experimental aortic surgery:Increased heat shock protein content is associated with ischemic tolerance of the spinal cord[J].Thoracic Cardiovascular Surgery,2002;124:162-170.
[13]De Bosscher,K Vanden,Berghe W H.Mechanisms of anti-inflammatory action and of immunosuppression by glucocorticoids:negative interference of activated glucocorticoid receptor with transcription factors[J].J Neuroimmunol,2000;109:16-22.
[14]Hartl FU.Molecular chaperones in celular protein folding[J].Nature,1996;381:571-579.
[15]Jakob U,Gaestel M,Engel K,et al.Small heat shock proteins are molecular chaperones[J].Journal of Biological Chemistry 1993;268:1517-1520.
[16]Lavoie JN,Hickey E,Weber LA,et al.Modulation of actin microfilament dynamics and fluid phase pinocytosis by phosphorylation of heat shock protein 27.Journal of Biological Chemistry 1993;32:24210-24214.
[17]van Noort JM.Stress proteins in CNS inflammation[J].Pathol,2008,214(2):267-275.
[18]Guay J,Lambert H,Gingras-Breton G,et al.Regulation of actin filament dynamics by p38 map kinase-mediated phosphorylation of heat shock protein 27.Journal of Cell Science 1997;110:357-368
[19]Huot J,Houle F,Spitz DR,et al.HSP27 phosphorylation-mediated resistance against actin fragmentation and cell death induced by oxidative stress[J].Cancer Research 1996;56:273-279,
[20]Geum D,Son GH,Kim K.Phosphorylation-dependent cellular localization and thermoprotective role of heat shock protein 25 in the hippocampal progenitor cells.Journal of Biological Chemistry 2002;277:19913-19921.
[21]Theriault JR,Lambert H,Chavez-Zobel AR,et al.Essential role of the NH2-terminal WD/EPF motif in the phosphorylation- activated protective function of mammalian Hsp27.Journal of Biological Chemistry 2004;279:23463-23471
[22]Tezel G,Wax MB.The mechanisms of hsp27 antibody mediated apoptosis in retinal neuronal cells.Journal of Neuroscience,2000;10:3552-3562
[23]刘文革,王锋,林佳俊,等.细胞凋亡在脊髓损伤中的表达及意义[J].福建医药杂志,2002;24(6):8.
[24]孙正义,赵斌,洪光祥,等.大鼠脊髓慢性压迫及减压后神经细胞凋亡及其相关基因的表达.中国脊柱脊髓杂志,2003;13(3):164.
[25]Mosser DD,Caron AW,Bourget L,et al.The chaperone function of hsp70 is required for protection against stress-induced apoptosis[J].Molecular Cell Biol 2000;20:7146-7159.
[26]Gabai VL,Mabuchi D,Mosser DD,et al.Hsp72 and stress kinase c-jun-N-terminal kinase regulate the bid-dependent pathway in tumor necrosis factor-induced apoptosis[J].Mol Cell Biology 2002;22:3415-3424.
[27]Paul C,Manero F,Gonin S,et al.Hsp27 as a negative regulator of cytochrome C release[J].Molecular Cell Biology 2002;22:816-834
[28]Benn SC,Perrelet D,Kato AC,et al.Hsp27 upregulation and phosphorylation is required for injured sensory and motor neuron survival[J].Neuron 2002;36:45-56.
[29]Bruey J-M,Ducasse C,Bonniaud P,et al.Hsp27 negatively regulates cell death by interacting with cytochrome C[J].Natural Cell Biology,2000;2:645-652.
[30]Saleh A,Srinivasula SM,Blakir L,et al.Negative regulation of the Apaf-1apoptosome by HSP70[J].Natural Cell Biology,2000;2:476-483.
[31]Helen M.Death versus survival:functional interaction between the apoptotic and stress-inducible heat shock protein pathways[J].Clin Invest,2005,115:2633-2639.
[32]Arya R,Mallik M,Lakhotia S C,et al.Heat shock genes -integrating cell survival and death[J].Biosci,2007;32(3):595-610.
[33]Adam R,Stankiewiczl,Guillaume,et al.HSP70 Inhibits Heat-induced Apoptosis Upstream of Mitochondria by Preventing Bax Translocation[J].Biolog Chem,2005;280(46):38729-38739.
[34]Basso A D,Solit D B,Chiosis G,et al.Akt forms an intracellular complex with heat shock protein 90(Hsp90) and Cdc37 and is destabilized by inhibitors of Hsp90 function[J].Biolog Chem,2002;277:39858-39866.
[35]Chan JY,Chang AY,Wang LL,et al.Protein kinase C-dependent mitochondrial translocation of proapoptotic protein Bax on activation of inducible nitric-oxide synthase in rostral ventrolateral medulla mediates cardiovascular depression during experimental endotoxemia[J].Mol Pharmacol,2007;71(4):1129-1139.
[36]毛定安,杨于嘉,虞佩兰,等.热休克蛋白70抑制大鼠颅脑损伤诱生型一氧化氮合酶mRNA的表达.中华实验外科杂志,2001;18:65-66.
[37]BruneB,Sandan K,yonKnethenA,et al.Apoptotic cell death and nitric oxide:activating and antagonistic transducing pathways[J].Biochemistry(Mosc),1998;63:817-825
[38]Chan JY,Cheng HL,Chou JL,et al.Heat shock protein 60 or 70 activates nitric-oxide synthase(NOS) Ⅰ- and inhibits NOS Ⅱ-associated signaling and depresses the mitochondrial apoptotic cascade during brain stem death[J].Biolog Chem,2007;282(7):4585-4600.
[39]傅勤,杜世新,傅永慧,等.神经生长因子和超氧化物歧化酶对急性脊髓损伤早期作用的实验研究.中国医科大学学报,2001;30(3)192.
[40]Turner CP,Panter SS,Sharp FR.Anti-oxidants prevent focal rat brain injury as assessed by induction of heat shock proteins following subarachnoid injections of lysed blood[J].Brain Res Mol Brain Res,1999;65:87-102.
[41]Yamashita H,Kawamata J,Okawa K,et al.Heat-shock protein 105 interacts with and suppresses aggregation of mutant Cu/Zn superoxide dismutase:clues to a possible strategy for treating ALS[J].Neurochem,2007;102(5):1497-1505.
[42]Pan PJ,Chan RC,Yang AH,et al.Protective effects of preconditioned local somatothermal stimulation on neuromuscular plasticity against ischemiareperfusion injury in rats[J].Orthop Res,2008;26(12):1670-1674.
[43]McCormick,Toomey D.Induction of heat shock proteins in humans using a clinically applicable model[J].British J Surgery Society,2003;90:620.
[44]Perdrizet,George A,Lena.Preoperative stress conditioning prevents paralysis after experimental aortic surgery:Increased heat shock protein content is associated with ischemic tolerance of the spinal cord[J].Thoracic Cardiovascular Surgery,2002;124:162-170.
[45]Sasara T,Cizkova D,Mestril R,et al.Spinal heat shock protein(70) expression:effect of spinal ischemia,hyperthermia(42℃)/hypothermia(27℃),NMDA receptor activation and potassium evoked depolarization on the induction[J].Neurochemistry International,2004;44:53-64.
[46]Cizkova D,Carmel JB,Yamamoto K,et al.Characerization of spinal HSP72induction and development of ischemic tolerance after spinal ischemia in rats[J].Experimental Neurology,2004;185:97-108.
[47]Selimoglu O,Ugurlucan M,Basaran M,et al.Efficacy of remote ischaemic preconditioning for spinal cord protection against ischaemic injury:association with heat shock protein expression[J].Folia Neuropathol,2008;46(3):204-212.
[48]Klass MG,Gavrikov V,Krishnamoorthy M,et al.Heat shock proteins,endothelin and peripheral neuronal injury[J].Neurosci Lett,2008,433(3):188-193.
[49]Bernadett K,Geoffrey B,Gerta V,et al.The Effect of Neonatal Nerve Injury on the Expression of Heat Shock Proteins in Developing Rat Motoneurones[J].Journal of Neurotrauma,2002;19(5):667-679.
[50]Hopkins DA,Plumier J,Currie RW.Induction of the 27-kDa heat shock protein (Hsp27) in the rat medulla oblongata after vagus nerve injury[J].Experimental Neurology,1998;153:173-183
[51]刘志彬,孔抗美,齐伟力.电针对大鼠慢性脊髓损伤热休克蛋白70及诱导型一氧化氮合酶表达的影响.中国临床康复,2004,8:5608-5609.
[52]Fleshner M,Campisi J,Amiri L,et al.Cat exposure induces both intra- and extracellular Hsp72:the role of adrenal hormjones[J].Psychoneuroendocrinology,2004;29:1142-1152.
[53]Zarah B,Josephine N,Heather D.Nonsteroidal anti-inflammatory drugs differentially affect the heat shock response in cultured spinal cord cells[J].Cell Stress &Chaperones,2005;10(3),185-196.
[54]Valen Guro,Tsutomu Kawakami,Peeter Tohepold,et al.Glucocorticoid pretreatment protects cardiac function and induces cardiac heat shock protein 72[J].Am J Physiol Heart Circ Physiol,2000;279:H836-H843.
[55]Lopez-Vales R,Garcia-Alias G,Fores J,et al.FK506 reduces tissue damage and prevents functional deficit after spinal cord injury in the rat[J].J Neurosci Res,2005;81:827-836.
[56]Gold BG,Voda J,Yu X,et al.The immunosuppressant FK506 elicits a neuronal heat shock response and protects against acrylamide neuropathy[J]. Experimental Neurology,2004;187:160-170.
[57]Guzman-Lenis MS,Vallejo C,Navarro X,et al.Analysis of FK506-mediated protection in an organotypic model of spinal cord damage:heat shock protein 70 levels are modulated in microglial cells[J].Neuroscience,2008;155(1):104-113.
[58]Ren M,Leng Y,Jeong M,et al.Valproic acid reduces brain damage induced by transient focal cerebral ischemia in rats:potential roles of histone deacetylase inhibition and heat shock protein induction.Journal of Neurochemistry 2004;89:1358-1367.
[59]Park SW,Yi TH,Miranpuri G,et al.Thiazolidinedione class of peroxisome proliferator-activated receptor gamma agonists prevents neuronal damage,motor dysfunction,myelin loss,neuropathic pain,and inflammation after spinal cord injury in adult rats[J].Pharmacol Exp Ther,2007;320(3):1002-1012.
[60]Sinn DI,Chu K,Lee ST,et al.Pharmacological induction of heat shock protein exerts neuroprotective effects in experimental intracerebral hemorrhage[J].Brain Res,2007;1135(1):167-176.
[61]Tavlor DM,Tradewell ML,Minotti S,et al.Characterizing the role of Hsp90 in production of heat shock proteins in motor neurons reveals a suppressive effect of wild-type Hsfl[J].Cell Stress Chaperones,2007;12(2):151-162.
[62]Suzuki K,Sawa Y,Kaneda Y,et al.In vivo gene transfection with heat shock protein 70 enhances myocardial tolerance to ischemia-reperfusion injury in rat[J].J Clin Invest,1997;99:1645-1650.
[63]Plumier J.-C,Krueger AM,Currie RW,et al.Transgenic mice expressing the human inducible HSP70 have hippocampal neurons resistant to ischemic injury[J].Cell Stress Chaperones,1997;2:162-167.
[64]Akbar MT,Lundber AMC,Vidyadaran S,et al.The neuroprotective effects of heat shock protein 27 overexpression in transgenic animals against kainate-induced seizures and hippocampal cell death[J].Journal of Biological Chemistry,2003;278:19956-19965.