兔脑缺血再灌注后NSE和S-100的表达及其血清水平的表化
|
摘要
目的探讨兔大脑中动脉缺血再灌注(MCAO／R)后脑组织神经元特异性烯醇化酶(NSE)和S-100蛋白的表达及其血清含量的变化。
方法将制作成功的兔MCAO／R模型58只，随机分为永久性缺血组30只和缺血再灌注组28只；另取10只动物行假手术分别作为缺血组(5只)及再灌注组(5只)的对照组。免疫组化法检测脑缺血再灌注不同时间脑组织NSE和S-100蛋白的表达，ELISA法检测血清NSE和S-100的含量。
结果假手术组脑组织NSE和S-100表达微弱，血清含量较低。永久性脑缺血3h和缺血再灌注2h后，脑组织NSE和S-100表达同步升高和降低、变化趋势基本一致，但NSE和S-100表达在缺血再灌注47h组较永久性缺血48h组再次明显升高。血清NSE和S-100水平直到缺血5h和再灌注5h开始同步升高，较脑组织NES和S-100表达时间延迟3h。
结论神经细胞和胶质细胞对缺血再灌注损伤非常敏感，再灌注可加重脑组织的损伤，破坏血脑屏障，NSE和S-100从脑脑脊液循环进入了血液循环，血清NSE和S-100的水平可作为评价脑损伤程度和转归的早期指标。
Objective To observe the expressions and serum levels of neuron-specific enolase(NSE) and S-100 after middle cerebral artery occlusion and reperfusion (MCAO/R) inrabbits.
Methods fifty-eight successful MCAO/R rabbit models were randomly divided intopermanent ischemic group 30 cases and ischemic reperfusion group 28 cases. Another10 rabbits were regarded as ischemic congtrol (5 cases) and reperfusion control (5cases). The expressions of NSE and S-100 in the brain tissue were determined withimmunochemical assay, and the serum levels of NSE and S-100 were measured withezyme linked immunosorbent assay (ELISA).
Results There were weak expressions and low serum levels of NSE and S-100 issham group. The expressions of NSE and S-100 increased/decreased and changedbasically as a same principles following permanent ischemic 3h and ischemicreperfusion 2h. But the expressions of NSE and S-100 in reperfusion 47h were alsohigh than that in permanent ischemic 48h. The serum levels of NSE and S-100increased same at ischemic 5h and reperfusion 5h, which were delayed 3h than that inbrain tissue expression.
Conclusion Neurons and glia were much sensitive to ischemic reperfusion. Thereperfusion might enhance brain tissue injury and destroy blood brain barrier to causethe NSE and S-100 into blood from cerebral spinal fluid, so that the serum levels ofNSE and S-100 might serve as early indexes evaluating brain injury and outcome.
方法将制作成功的兔MCAO／R模型58只，随机分为永久性缺血组30只和缺血再灌注组28只；另取10只动物行假手术分别作为缺血组(5只)及再灌注组(5只)的对照组。免疫组化法检测脑缺血再灌注不同时间脑组织NSE和S-100蛋白的表达，ELISA法检测血清NSE和S-100的含量。
结果假手术组脑组织NSE和S-100表达微弱，血清含量较低。永久性脑缺血3h和缺血再灌注2h后，脑组织NSE和S-100表达同步升高和降低、变化趋势基本一致，但NSE和S-100表达在缺血再灌注47h组较永久性缺血48h组再次明显升高。血清NSE和S-100水平直到缺血5h和再灌注5h开始同步升高，较脑组织NES和S-100表达时间延迟3h。
结论神经细胞和胶质细胞对缺血再灌注损伤非常敏感，再灌注可加重脑组织的损伤，破坏血脑屏障，NSE和S-100从脑脑脊液循环进入了血液循环，血清NSE和S-100的水平可作为评价脑损伤程度和转归的早期指标。
Objective To observe the expressions and serum levels of neuron-specific enolase(NSE) and S-100 after middle cerebral artery occlusion and reperfusion (MCAO/R) inrabbits.
Methods fifty-eight successful MCAO/R rabbit models were randomly divided intopermanent ischemic group 30 cases and ischemic reperfusion group 28 cases. Another10 rabbits were regarded as ischemic congtrol (5 cases) and reperfusion control (5cases). The expressions of NSE and S-100 in the brain tissue were determined withimmunochemical assay, and the serum levels of NSE and S-100 were measured withezyme linked immunosorbent assay (ELISA).
Results There were weak expressions and low serum levels of NSE and S-100 issham group. The expressions of NSE and S-100 increased/decreased and changedbasically as a same principles following permanent ischemic 3h and ischemicreperfusion 2h. But the expressions of NSE and S-100 in reperfusion 47h were alsohigh than that in permanent ischemic 48h. The serum levels of NSE and S-100increased same at ischemic 5h and reperfusion 5h, which were delayed 3h than that inbrain tissue expression.
Conclusion Neurons and glia were much sensitive to ischemic reperfusion. Thereperfusion might enhance brain tissue injury and destroy blood brain barrier to causethe NSE and S-100 into blood from cerebral spinal fluid, so that the serum levels ofNSE and S-100 might serve as early indexes evaluating brain injury and outcome.
引文
[1] Butterworth RJ, Wassif WS, Sherwood RA, et al. Serum neuron-specific enolase, carnosinase, and their ratio in acute sstoke[J]. Stroke, 1996, 27(11):2064-2068.
[2] Abrah HD, Butterworth RJ, Bath PM, et al. Serum S-100 protein: relationship to clinical outcome in acute stroke[J]. Ann Clin Biochem, 1997, 34(4):366-370.
[3] 贺勇，尤志瑶，周少华，等．短暂性脑缺血发作患者血清S100和神经元特异性烯醇化酶的测定及临床意义[J]．实用医学杂志，2005，21(1)：37-38．
[4] 王晓明，张国元，许可，等．脑梗死患者血清神经元特异性烯醇化酶、S-100B蛋白和髓鞘碱性蛋白含量的变化[J]．中国危重病急救医学，2005，17(9)：572-573．
[5] 孔令琦，谢敬霞，韩鸿宾．提高线栓法大脑中动脉闭塞性兔脑缺血模型稳定性和可重复性的研究[J]．中国医学影像技术，2004，20(2)：209-212．
[6] Lafon-Cazal M, Bougauh I, Steinberg G, et al. Measurement of gamma enolase, a new method for selective quantification of neurotoxicity independentlyfrom gliallysis[J]. Brain Res, 1992, 593(1):63-68.
[7] Pleines UE, Morganti-Kossmann MC, Rancan M, et al. S-100 beta refleets the extent of injury and outcome, whereas neuronal specific enolase is a better indicator of neuroinflammation in patients with severtraumatic brain injury[1]. J Neumtraurma, 2001, 18(5):491-498.
[8] 杨慧君，王生池，李云萍，等．急性脑梗死神经元特异性烯醇化酶的临床意义[J]．中华实用医学，2004，6(4)：16-18．
[9]. Moroz OV, Antson AA, Murshudov GN, et al. The three dimensional structure of human S100A12. Acta Crystallogr D Biol Crystallogr, 2001, 57: 20-29
[10] Ridinger K, Schafer BW, Durussel I, et al. S100A13. Biochemical characterization and subcellular localization in different cell lines. J Biol Chem, 2000; 275(12): 8686-94
[11] Hardemark HG, Ericsson N, Kotwica Z, et al. S100 protein and neuron specific enolase in CSF after experimental traumatic or focal ischemic brain damage [J]. J Neurosurg, 1999; 71:727
[12] Marangos PJ. Neuron specific enolase, a clinically useful marker for neurons and neuroendocrine cells [J] . Ann Rev Neurosci, 1987,10:269.
[13] Angelov DN, Neiss WF, Gunkel A, et al. Axotomy induces intranuclear immunolocalization of neuron-specific enolase in facial and hypoglossal neurons of the rat [J] . J Neurocyto, 1994,23:218.
[14] Lamers KJ, van Engelen BG, Gabreels FJ, et al. Cerebrospinal neuron-specific enolase, S-100 and myelin basic protein in neurological disorders [J] . Acta Neurol Scand, 1995,92:247.
[15] Kaneta M, Aoki T, Onagi H, et al. Biochemical and immunochemical properties of modified human neuron-specific enolase [J] . Biol Bull, 1997,20:556.
[16] Gross J, Ungethum U, Andreeva N, et al. Glutamate-induced efflux of protein, neuron-specific enolase and lactate dehydrogenase from a mesencephalic cell culture [J] . Eur J Clin Chem Clin Biochem, 1996,34:305.
[17] Wunderlich MT, Ebert AD, Kratz T, et al. Early neurobehavioral outcome after stroke is release of neurobiochemical markers of brain damage[J]. Stroke, 1999,30(6):1190-1995.
[18] 王粤，张红，龚薇薇，等．脑缺血再灌注损伤后NSE和S-100β的表达[J]．齐鲁医学杂志，2004，19(1)：1-3．
[19] Angelov DN, Neiss WF, Gunkel A, et al. Axotomy induces intranuclear immunolocalization of neuron-specific enotase in facial and hypoglossal neurons of the rat [J]. J Neurocyto, 1994, 23:218.
[20] Lamers KJ, van Engelen BG, Cabreels FJ, et al. Cerebrospinal neuron-specific enolase, S-100 and myelin basic protein in neurological disorders [J]. Acta Neurol Scand, 1995, 92:247.
[21] Kaneta M, Aoki T, Onagi H, et al. Biochemical and immunochemical properties of modified human neuron-specific enolase [J]. Biol Bull, 1997, 20:556
[22] Rustandi RR, Baldisseri DM, Weber DJ. Structure of the negative regulatory domain of p53 bound to S-100β(beta)[J]. Nat Struct Biol, 2000, 7(7):570-574.
[23] 张晓伟．S-100蛋白研究进展[J]．沪州医学院学报，2005，28(3)：270-273
[1] Butterworth RJ, Wassif WS, Sherwood RA, et al. Serum neuron-specific enolase, carnosinase, and their ratio in acute sstoke[J]. Stroke, 1996, 27(11):2064-2068.
[2] Abrah HD, Butterworth RJ, Bath PM, et al. Serum S-100 protein: relationship to clinical outcome in acute stroke[J]. Ann Clin Biochem, 1997, 34(4):366-370.
[3] Lafon-Cazal M, Bougauh I, Steinberg G, et al. Measurement of gamma enolase, a new method for selective quantification of neurotoxicity independentlyfrom gliallysis[J]. Brain Res, 1992, 593(1):63-68.
[4] Pleines UE, Morganti-Kossmann MC, Rancan M, et al. S-100 beta refleets the extent of injury and outcome, whereas neuronal specific enolase is a better indicator of neuroinflammation in patients with severtraumatic brain injury[J]. J Neumtraurma, 2001, 18(5):491-498.
[5] 杨慧君，王生池，李云萍，等．急性脑梗死神经元特异性烯醇化酶的临床意义[J]．中华实用医学，2004，6(4)：16-18．
[6] Ozumi I, Chiu FC, Norton WT. Biochemical and immunocytochemical changes in glial fibrillary acidic protein after stab wounds[J]. Brain Res, 1990: 524:64
[7] Pasquale AC, Giordana MT, Migheli A, et al. Glial fibrillary acidic protein after experimental cerebral injury [J]. J Neuropathol Exp Neurol, 2001:51:560
[8]Calvo JL, Carbonell AL, Boya J. Co-expression of glial fivrillary acidic protein and vimentin in teactive astrocytes following brain injury in rats [J]. Brain Res, 1997; 566:333.
[9]Eng LF. Regulation of glial intermediate filaments in astrogliosis, The Biochemical Pathology of astrocytes, Neurology and Neurobiology, 1998; 39:79
[10] Moroz 0V, Antson AA, Murshudov GN, et al. The three dimensional structure of human S100A12. Acta Crystallogr D Biol Crystallogr,. 2001, 57: 20-29
[ll]Ridinger K, Schafer BW, Durussel I, et al. S100A13. Biochemical characterization and subcellular localization in different cell lines. J Biol Chem, 2000; 275(12): 8686-94
[12] Hessian PA, Fisher L The heterodimeric complex of MRP-8 (S100A8) and MRP-14 (S100A9). Antibody recognition, epitope definition and the implications for structure. Eur J Biochem, 2001, 268(2): 353-63
[13] Giorgio F, Silvia B, Stefania F et al. The s100: A protein family in the serch of a function. Progress in Neurobiology, 1995, 46:71-82
[14] Beat WS, Claus WH. The S100 family of EF-hand calcium binding proteins: functions and pathology. Trends in Biochem Sci, 1996, 21:134-140
[15] Ridinger K, Schafer BW, Durussel I, et al. S100A13. Biochemical characterization and subcellular localization in different cell lines. J Biol Chem, 2000, 275(12): 8686-94
[16] Taggart DP, Mazel JW, Meston N, et al. Comparison of serum S100B levels during CABG and intracardiac operations. Ann Thorac Surg, 1997, 63:492-496
[17] Herrmann M, Ebert AD, Galazky I, et al. Neurobehavioral outcome prediction after cardiac surgery, role of neurobiochemical markers of damage to neuronal and glial brain tissue. Stroke; 2000, 31:645-650
[18] Buttner T, Weyers S, Postert T, et al. S100 protein:serum marker of focal brain damage after ischemic territorial MCA infarction. Stroke, 1997, 28:1961-1965
[19] Persson L, Hardemark HG, Gustafsson J, et al. S100 protein and neuron specific enolase in cerebrospinal fluid and serum: markers of cell damage in human central nervous system. Stroke, 1997, 18:911-918
[20] Woertgen C, Rothoerl RD, Holzschuh M, et al. Comparison of serial S100 and NSE serum measurements after severe head injury. Acta Neurochirurgica, 1997, 139:1161-1165
[21] Ingebrigtsen T, Romner B. Serial S100 protein serum measurements related to early magnetic resonance imaging after minor head injury. J Neurosurg, 1996, 85:945-948
[22]Bidmon B, et al. Kidney Int 2000, 58(6) :2400—2407
[23] Heizmann CW, Cox, JA. New perspectives on S100 proteins: a multi functional Ca(2+) , Zn(2+) and Cu(2+) binding protein family. Biometals, 1998, 11(4): 383-97
[24] Hauschild A, Engel G, Brenner W, et al. Predictive value of serum S100B for monitoring patients with metastatic melanoma during chemotherapy and/or immunotherapy. Br J Dermatol, 1999, 140(6): 1065-71
[25] BrownIR, RushSJ, Ivy GO. Induction of a heat shock gene at the site of tissue injury in the rat brain[J].Neuron, 1999; 2:1559
[26] Posmantur R, Hayes RL, Dixon CE, et al. Neurofilament 68 and neurofilament200 protein levels decrease after traumatic brain injury[J]. J Neurotrauma, 1998; 11(5):533-545
[27] Higashi KJ. UOEH 1999, 21(2) : 157～169
[28] Kropp S, Zerr I, Schaeffer SWJ, et al.Increased of neuron-specific enotase in patients with Creutzfeldt-Jakob disease [J]. Neurosci Lett, 1999, 261 : 124.
[29] Elimian A, Figueroa R, Verma U, et al.Amniotic fluid neuron-specific enolase:a role in predicting neonatal neurologic injury [J] ?Obstet Gynecol, 1998, 92:546.
[30] 吴旭，王德文．定量检测NSE和S2100推断脑挫伤时间的动物实验[J]．中国法医学杂志，1999；14：141～144
[31] 陈龙，张其英．纤维连接蛋白与皮肤创伤的研究[J]．中国法医学杂志，1999；14(3)：185～187
[32] 陈龙，陈忆九．大鼠脑挫伤后FN表达与损伤时程关系的研究[J]．法医学杂志，200l；17(1)：1～3
[33] Snyder SH. Nitric oxide. No endothelial [J]. Nature, 1995; 377:196
[34] Murphy S, Simmons ML Agullo L, et al. Synthesis of nitric oxide in CNS glial cells [J]. Trends Neuroscl, 1993; 16(8):323
[35] Zhang ZG, Chopp M, Gautam S, et al. Up regulation of neuronal nitric oxide synthase and mRNA, and selectives paring of nitricoxide synthase containing eurons after focal cerebral ischemia in rat[J]. Brain Res, 1994; 654(1):85
[36] Zimmernan J J, Kochanek P M, Meadow W, etal. Molecular pathophysiology in critical careillness[J].Crit Care Med, 1993; 21:400
[37] Chen S, Aston J G.Cerebellar injury induces NADPH diaphorase In purkinje and inferior olivary neurons in the rat[J]. Exp Neurol, 1994; 126(2):270
[38] Riossa F. Experimentia. 1962, 18:51.
[39]．周力文等．中华麻醉学杂志1999，2：118～119．
[40] William J. Physiological Reviews 1992, 72(4):1063.
[41]. Mallouk Y, et al. Int J Mol Med 1999, 4(5):463～474.
[43] Coronato S, et al. Medicina (B Aires) 1999, 59:477～486.
[44] Lepore DA, et al. Muscle Nerve 2000, 23(12):1847～1855.
[45] Kumem, et al. J Lab Clin Med 1996, 128:251～253.
[46] Jayakamar J, et al. Circulation 2000, 102(19 Suppl 3):Ⅲ302～306.
[47] Villar J, et al. CrAt Care Med 1994, 22:914.
[48] Gass p, Priorp k. Neuroscience 1995, 65(1)27～36.
[49]. Yang YL, Lin MT. Am J Physiol 1999, 276(6 Pt 2).
[50]Wallach D. TIBIS 1997, 22; 107-109.
[51].Kamadn S, et al. Oncogene 1997,15:285—290.
[52].Mosser DD, et al. Mol Cell Biol 2000, 20(19) :7146—7159. .
[53].Okubo A, Kinouchi H, Owada Y, ET AL. Brain Res Mol Brain Res 2000, 8:84(1-2): 127-134.
[54]Tomimoto H, Takemoto O, Akiguchi I, et al. Acta Neuropathol (Berl) 1999,97:22-30.
[55] Quinn GB, Reeves IG, Day IN. Mapping of antigenic sites in human neuron-specific enolase by expression subcloning [J] . Clin Chem, 1994,40:790.
[56] Marangos PJ. Neuron specific enolase, a clinically useful marker for neurons and neuroendocrine cells [J] . Ann Rev Neurosci, 1987,10:269.
[57] Angelov DN, Neiss WF, Gunkel A, et al. Axotomy induces intranuclear immunolocalization of neuron-specific enolase in facial and hypoglossal neurons of the rat [J] . J Neurocyto, 1994,23:218.
[58] Lamers KJ, van Engelen BG, Gabreels FJ, et al. Cerebrospinal neuron-specific enolase, S-100 and myelin basic protein in neurological disorders [J] . Acta Neurol Scand, 1995,92:247.
[59] Kaneta M, Aoki T, Onagi H, et al. Biochemical and immunochemical properties of modified human neuron-specific enolase [J] . Biol Bull, 1997,20:556.
[60] Missler U, Wiesmann M, Friedrich C, et al. S-100 protein and neuron-specific enolase concentration in blood as indicators of infarction volume and progrosis in acute ischemic stroke [J] . Stroke, 1997,28:1956.
[61] Schaarschmidt H, Prange HW, Reiber H. Neuron-specific enolase concentrations in blood as a progrostic parameter in cerebrovascular diseases [ J] . Stroke, 1994,25:558.
[62] Barone FC, Clark RK, Price WJ, et al. Neuron-specific enolase increases in cerebral and systemic circulation following focal ischemia [J] . Brain Res, 1993,623:77.
[63] Correale J,Rabinowics AL,Heck CN,et al.Status epilepticus increases CSF levels of neuron-specific enolase and alters the blood-brain barrier [J] .Neurology,1998,50:1388.
[64] Gross J, Ungethum U, Andreeva N, et al. Glutamate-induced efflux of protein, neuron-specific enolase and lactate dehydrogenase from a mesencephalic cell culture [J] . Eur J Clin Chem Clin Biochem, 1996,34:305.
[65] Wunderlich MT,Ebert AD,Kratz T,et al.Early neurobehavioral outcome after stroke is related to release of neurobiochemical markers of brain damage [J] .Stroke, 1999,30:1190.
[66] Sanker R, Shin DH, Wasterlain CG. Serum neuron-specific enolase is a marker for neuronal damage following status epilepticus in the rat [ J] . Epilepsy Res, 1997,28:129.
[67] DeGiorgio CM,Heck CN,Rabinowicz AL,et al.Serum neuron-specific enolase in the major subtypes of status epilepticus [J] .Neurology, 1999,52:746.
[68] Yamazaki Y, Yada K, Morii S, et al. Diagnostic significance of serum-specific enolase and myelin basic protein assay in patients with acute head injury [J] . SurgNeurol, 1995,43:267.
[69] Baler R, Dahl G, Voellmy R. Activation of human heat shock genes is accompanied by oligomerization, modification, and rapid trpid translocation of heat shock transcription fact HSF1[J].MolCell Biol., 1993, 13: 2 486
[70] Cowling DW, James CA, Bardwell EA, et al. Consensus sequence for Eschcrichia coli heat shock gene promoters. USA Proc Nail Sci., 1985, S2:2 679～2 683
[71] GradyMS, Mclaughlin MR, Christman CW, et al. The use of antibodies targeted against the Neurofilament subunits for the detection of diffuse axonal injury in human[J].Jneuropathol ExpNeurol, 1997: 52:143
[72] Nakamura M, Raghupathi R, Murry DE, et al. Over expression of bcl-2 is neuroprotective after experimental brain injury in transgenic mice [J]. Jcomp Neurol, 1999:412(4):681～692
[73] 陈龙，陈忆九．大鼠脑挫伤后组织学及BAX／bcl22表达的研究[J]．法医学杂志，2000；16(4)：211～213
[74] 贺立文，祝家镇．细胞因子在损伤时间中原位杂交法研究[J]．中国法医学杂志，1997；12(3)：133～136
[2] Abrah HD, Butterworth RJ, Bath PM, et al. Serum S-100 protein: relationship to clinical outcome in acute stroke[J]. Ann Clin Biochem, 1997, 34(4):366-370.
[3] 贺勇，尤志瑶，周少华，等．短暂性脑缺血发作患者血清S100和神经元特异性烯醇化酶的测定及临床意义[J]．实用医学杂志，2005，21(1)：37-38．
[4] 王晓明，张国元，许可，等．脑梗死患者血清神经元特异性烯醇化酶、S-100B蛋白和髓鞘碱性蛋白含量的变化[J]．中国危重病急救医学，2005，17(9)：572-573．
[5] 孔令琦，谢敬霞，韩鸿宾．提高线栓法大脑中动脉闭塞性兔脑缺血模型稳定性和可重复性的研究[J]．中国医学影像技术，2004，20(2)：209-212．
[6] Lafon-Cazal M, Bougauh I, Steinberg G, et al. Measurement of gamma enolase, a new method for selective quantification of neurotoxicity independentlyfrom gliallysis[J]. Brain Res, 1992, 593(1):63-68.
[7] Pleines UE, Morganti-Kossmann MC, Rancan M, et al. S-100 beta refleets the extent of injury and outcome, whereas neuronal specific enolase is a better indicator of neuroinflammation in patients with severtraumatic brain injury[1]. J Neumtraurma, 2001, 18(5):491-498.
[8] 杨慧君，王生池，李云萍，等．急性脑梗死神经元特异性烯醇化酶的临床意义[J]．中华实用医学，2004，6(4)：16-18．
[9]. Moroz OV, Antson AA, Murshudov GN, et al. The three dimensional structure of human S100A12. Acta Crystallogr D Biol Crystallogr, 2001, 57: 20-29
[10] Ridinger K, Schafer BW, Durussel I, et al. S100A13. Biochemical characterization and subcellular localization in different cell lines. J Biol Chem, 2000; 275(12): 8686-94
[11] Hardemark HG, Ericsson N, Kotwica Z, et al. S100 protein and neuron specific enolase in CSF after experimental traumatic or focal ischemic brain damage [J]. J Neurosurg, 1999; 71:727
[12] Marangos PJ. Neuron specific enolase, a clinically useful marker for neurons and neuroendocrine cells [J] . Ann Rev Neurosci, 1987,10:269.
[13] Angelov DN, Neiss WF, Gunkel A, et al. Axotomy induces intranuclear immunolocalization of neuron-specific enolase in facial and hypoglossal neurons of the rat [J] . J Neurocyto, 1994,23:218.
[14] Lamers KJ, van Engelen BG, Gabreels FJ, et al. Cerebrospinal neuron-specific enolase, S-100 and myelin basic protein in neurological disorders [J] . Acta Neurol Scand, 1995,92:247.
[15] Kaneta M, Aoki T, Onagi H, et al. Biochemical and immunochemical properties of modified human neuron-specific enolase [J] . Biol Bull, 1997,20:556.
[16] Gross J, Ungethum U, Andreeva N, et al. Glutamate-induced efflux of protein, neuron-specific enolase and lactate dehydrogenase from a mesencephalic cell culture [J] . Eur J Clin Chem Clin Biochem, 1996,34:305.
[17] Wunderlich MT, Ebert AD, Kratz T, et al. Early neurobehavioral outcome after stroke is release of neurobiochemical markers of brain damage[J]. Stroke, 1999,30(6):1190-1995.
[18] 王粤，张红，龚薇薇，等．脑缺血再灌注损伤后NSE和S-100β的表达[J]．齐鲁医学杂志，2004，19(1)：1-3．
[19] Angelov DN, Neiss WF, Gunkel A, et al. Axotomy induces intranuclear immunolocalization of neuron-specific enotase in facial and hypoglossal neurons of the rat [J]. J Neurocyto, 1994, 23:218.
[20] Lamers KJ, van Engelen BG, Cabreels FJ, et al. Cerebrospinal neuron-specific enolase, S-100 and myelin basic protein in neurological disorders [J]. Acta Neurol Scand, 1995, 92:247.
[21] Kaneta M, Aoki T, Onagi H, et al. Biochemical and immunochemical properties of modified human neuron-specific enolase [J]. Biol Bull, 1997, 20:556
[22] Rustandi RR, Baldisseri DM, Weber DJ. Structure of the negative regulatory domain of p53 bound to S-100β(beta)[J]. Nat Struct Biol, 2000, 7(7):570-574.
[23] 张晓伟．S-100蛋白研究进展[J]．沪州医学院学报，2005，28(3)：270-273
[1] Butterworth RJ, Wassif WS, Sherwood RA, et al. Serum neuron-specific enolase, carnosinase, and their ratio in acute sstoke[J]. Stroke, 1996, 27(11):2064-2068.
[2] Abrah HD, Butterworth RJ, Bath PM, et al. Serum S-100 protein: relationship to clinical outcome in acute stroke[J]. Ann Clin Biochem, 1997, 34(4):366-370.
[3] Lafon-Cazal M, Bougauh I, Steinberg G, et al. Measurement of gamma enolase, a new method for selective quantification of neurotoxicity independentlyfrom gliallysis[J]. Brain Res, 1992, 593(1):63-68.
[4] Pleines UE, Morganti-Kossmann MC, Rancan M, et al. S-100 beta refleets the extent of injury and outcome, whereas neuronal specific enolase is a better indicator of neuroinflammation in patients with severtraumatic brain injury[J]. J Neumtraurma, 2001, 18(5):491-498.
[5] 杨慧君，王生池，李云萍，等．急性脑梗死神经元特异性烯醇化酶的临床意义[J]．中华实用医学，2004，6(4)：16-18．
[6] Ozumi I, Chiu FC, Norton WT. Biochemical and immunocytochemical changes in glial fibrillary acidic protein after stab wounds[J]. Brain Res, 1990: 524:64
[7] Pasquale AC, Giordana MT, Migheli A, et al. Glial fibrillary acidic protein after experimental cerebral injury [J]. J Neuropathol Exp Neurol, 2001:51:560
[8]Calvo JL, Carbonell AL, Boya J. Co-expression of glial fivrillary acidic protein and vimentin in teactive astrocytes following brain injury in rats [J]. Brain Res, 1997; 566:333.
[9]Eng LF. Regulation of glial intermediate filaments in astrogliosis, The Biochemical Pathology of astrocytes, Neurology and Neurobiology, 1998; 39:79
[10] Moroz 0V, Antson AA, Murshudov GN, et al. The three dimensional structure of human S100A12. Acta Crystallogr D Biol Crystallogr,. 2001, 57: 20-29
[ll]Ridinger K, Schafer BW, Durussel I, et al. S100A13. Biochemical characterization and subcellular localization in different cell lines. J Biol Chem, 2000; 275(12): 8686-94
[12] Hessian PA, Fisher L The heterodimeric complex of MRP-8 (S100A8) and MRP-14 (S100A9). Antibody recognition, epitope definition and the implications for structure. Eur J Biochem, 2001, 268(2): 353-63
[13] Giorgio F, Silvia B, Stefania F et al. The s100: A protein family in the serch of a function. Progress in Neurobiology, 1995, 46:71-82
[14] Beat WS, Claus WH. The S100 family of EF-hand calcium binding proteins: functions and pathology. Trends in Biochem Sci, 1996, 21:134-140
[15] Ridinger K, Schafer BW, Durussel I, et al. S100A13. Biochemical characterization and subcellular localization in different cell lines. J Biol Chem, 2000, 275(12): 8686-94
[16] Taggart DP, Mazel JW, Meston N, et al. Comparison of serum S100B levels during CABG and intracardiac operations. Ann Thorac Surg, 1997, 63:492-496
[17] Herrmann M, Ebert AD, Galazky I, et al. Neurobehavioral outcome prediction after cardiac surgery, role of neurobiochemical markers of damage to neuronal and glial brain tissue. Stroke; 2000, 31:645-650
[18] Buttner T, Weyers S, Postert T, et al. S100 protein:serum marker of focal brain damage after ischemic territorial MCA infarction. Stroke, 1997, 28:1961-1965
[19] Persson L, Hardemark HG, Gustafsson J, et al. S100 protein and neuron specific enolase in cerebrospinal fluid and serum: markers of cell damage in human central nervous system. Stroke, 1997, 18:911-918
[20] Woertgen C, Rothoerl RD, Holzschuh M, et al. Comparison of serial S100 and NSE serum measurements after severe head injury. Acta Neurochirurgica, 1997, 139:1161-1165
[21] Ingebrigtsen T, Romner B. Serial S100 protein serum measurements related to early magnetic resonance imaging after minor head injury. J Neurosurg, 1996, 85:945-948
[22]Bidmon B, et al. Kidney Int 2000, 58(6) :2400—2407
[23] Heizmann CW, Cox, JA. New perspectives on S100 proteins: a multi functional Ca(2+) , Zn(2+) and Cu(2+) binding protein family. Biometals, 1998, 11(4): 383-97
[24] Hauschild A, Engel G, Brenner W, et al. Predictive value of serum S100B for monitoring patients with metastatic melanoma during chemotherapy and/or immunotherapy. Br J Dermatol, 1999, 140(6): 1065-71
[25] BrownIR, RushSJ, Ivy GO. Induction of a heat shock gene at the site of tissue injury in the rat brain[J].Neuron, 1999; 2:1559
[26] Posmantur R, Hayes RL, Dixon CE, et al. Neurofilament 68 and neurofilament200 protein levels decrease after traumatic brain injury[J]. J Neurotrauma, 1998; 11(5):533-545
[27] Higashi KJ. UOEH 1999, 21(2) : 157～169
[28] Kropp S, Zerr I, Schaeffer SWJ, et al.Increased of neuron-specific enotase in patients with Creutzfeldt-Jakob disease [J]. Neurosci Lett, 1999, 261 : 124.
[29] Elimian A, Figueroa R, Verma U, et al.Amniotic fluid neuron-specific enolase:a role in predicting neonatal neurologic injury [J] ?Obstet Gynecol, 1998, 92:546.
[30] 吴旭，王德文．定量检测NSE和S2100推断脑挫伤时间的动物实验[J]．中国法医学杂志，1999；14：141～144
[31] 陈龙，张其英．纤维连接蛋白与皮肤创伤的研究[J]．中国法医学杂志，1999；14(3)：185～187
[32] 陈龙，陈忆九．大鼠脑挫伤后FN表达与损伤时程关系的研究[J]．法医学杂志，200l；17(1)：1～3
[33] Snyder SH. Nitric oxide. No endothelial [J]. Nature, 1995; 377:196
[34] Murphy S, Simmons ML Agullo L, et al. Synthesis of nitric oxide in CNS glial cells [J]. Trends Neuroscl, 1993; 16(8):323
[35] Zhang ZG, Chopp M, Gautam S, et al. Up regulation of neuronal nitric oxide synthase and mRNA, and selectives paring of nitricoxide synthase containing eurons after focal cerebral ischemia in rat[J]. Brain Res, 1994; 654(1):85
[36] Zimmernan J J, Kochanek P M, Meadow W, etal. Molecular pathophysiology in critical careillness[J].Crit Care Med, 1993; 21:400
[37] Chen S, Aston J G.Cerebellar injury induces NADPH diaphorase In purkinje and inferior olivary neurons in the rat[J]. Exp Neurol, 1994; 126(2):270
[38] Riossa F. Experimentia. 1962, 18:51.
[39]．周力文等．中华麻醉学杂志1999，2：118～119．
[40] William J. Physiological Reviews 1992, 72(4):1063.
[41]. Mallouk Y, et al. Int J Mol Med 1999, 4(5):463～474.
[43] Coronato S, et al. Medicina (B Aires) 1999, 59:477～486.
[44] Lepore DA, et al. Muscle Nerve 2000, 23(12):1847～1855.
[45] Kumem, et al. J Lab Clin Med 1996, 128:251～253.
[46] Jayakamar J, et al. Circulation 2000, 102(19 Suppl 3):Ⅲ302～306.
[47] Villar J, et al. CrAt Care Med 1994, 22:914.
[48] Gass p, Priorp k. Neuroscience 1995, 65(1)27～36.
[49]. Yang YL, Lin MT. Am J Physiol 1999, 276(6 Pt 2).
[50]Wallach D. TIBIS 1997, 22; 107-109.
[51].Kamadn S, et al. Oncogene 1997,15:285—290.
[52].Mosser DD, et al. Mol Cell Biol 2000, 20(19) :7146—7159. .
[53].Okubo A, Kinouchi H, Owada Y, ET AL. Brain Res Mol Brain Res 2000, 8:84(1-2): 127-134.
[54]Tomimoto H, Takemoto O, Akiguchi I, et al. Acta Neuropathol (Berl) 1999,97:22-30.
[55] Quinn GB, Reeves IG, Day IN. Mapping of antigenic sites in human neuron-specific enolase by expression subcloning [J] . Clin Chem, 1994,40:790.
[56] Marangos PJ. Neuron specific enolase, a clinically useful marker for neurons and neuroendocrine cells [J] . Ann Rev Neurosci, 1987,10:269.
[57] Angelov DN, Neiss WF, Gunkel A, et al. Axotomy induces intranuclear immunolocalization of neuron-specific enolase in facial and hypoglossal neurons of the rat [J] . J Neurocyto, 1994,23:218.
[58] Lamers KJ, van Engelen BG, Gabreels FJ, et al. Cerebrospinal neuron-specific enolase, S-100 and myelin basic protein in neurological disorders [J] . Acta Neurol Scand, 1995,92:247.
[59] Kaneta M, Aoki T, Onagi H, et al. Biochemical and immunochemical properties of modified human neuron-specific enolase [J] . Biol Bull, 1997,20:556.
[60] Missler U, Wiesmann M, Friedrich C, et al. S-100 protein and neuron-specific enolase concentration in blood as indicators of infarction volume and progrosis in acute ischemic stroke [J] . Stroke, 1997,28:1956.
[61] Schaarschmidt H, Prange HW, Reiber H. Neuron-specific enolase concentrations in blood as a progrostic parameter in cerebrovascular diseases [ J] . Stroke, 1994,25:558.
[62] Barone FC, Clark RK, Price WJ, et al. Neuron-specific enolase increases in cerebral and systemic circulation following focal ischemia [J] . Brain Res, 1993,623:77.
[63] Correale J,Rabinowics AL,Heck CN,et al.Status epilepticus increases CSF levels of neuron-specific enolase and alters the blood-brain barrier [J] .Neurology,1998,50:1388.
[64] Gross J, Ungethum U, Andreeva N, et al. Glutamate-induced efflux of protein, neuron-specific enolase and lactate dehydrogenase from a mesencephalic cell culture [J] . Eur J Clin Chem Clin Biochem, 1996,34:305.
[65] Wunderlich MT,Ebert AD,Kratz T,et al.Early neurobehavioral outcome after stroke is related to release of neurobiochemical markers of brain damage [J] .Stroke, 1999,30:1190.
[66] Sanker R, Shin DH, Wasterlain CG. Serum neuron-specific enolase is a marker for neuronal damage following status epilepticus in the rat [ J] . Epilepsy Res, 1997,28:129.
[67] DeGiorgio CM,Heck CN,Rabinowicz AL,et al.Serum neuron-specific enolase in the major subtypes of status epilepticus [J] .Neurology, 1999,52:746.
[68] Yamazaki Y, Yada K, Morii S, et al. Diagnostic significance of serum-specific enolase and myelin basic protein assay in patients with acute head injury [J] . SurgNeurol, 1995,43:267.
[69] Baler R, Dahl G, Voellmy R. Activation of human heat shock genes is accompanied by oligomerization, modification, and rapid trpid translocation of heat shock transcription fact HSF1[J].MolCell Biol., 1993, 13: 2 486
[70] Cowling DW, James CA, Bardwell EA, et al. Consensus sequence for Eschcrichia coli heat shock gene promoters. USA Proc Nail Sci., 1985, S2:2 679～2 683
[71] GradyMS, Mclaughlin MR, Christman CW, et al. The use of antibodies targeted against the Neurofilament subunits for the detection of diffuse axonal injury in human[J].Jneuropathol ExpNeurol, 1997: 52:143
[72] Nakamura M, Raghupathi R, Murry DE, et al. Over expression of bcl-2 is neuroprotective after experimental brain injury in transgenic mice [J]. Jcomp Neurol, 1999:412(4):681～692
[73] 陈龙，陈忆九．大鼠脑挫伤后组织学及BAX／bcl22表达的研究[J]．法医学杂志，2000；16(4)：211～213
[74] 贺立文，祝家镇．细胞因子在损伤时间中原位杂交法研究[J]．中国法医学杂志，1997；12(3)：133～136