慢性脑缺血大鼠脑组织IL-1β、IL-6、TNF-α及β-淀粉样蛋白1-42的表达研究
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摘要
目的研究慢性脑缺血大鼠脑组织IL-1β、IL-6、TNF-α等免疫炎性因子和β-淀粉样蛋白1-42(Aβ1-42)表达水平的变化及两者的相关性。方法将16只SD大鼠采用随机数字表法分为模型组和假手术组,各8只。模型组大鼠永久结扎双侧颈总动脉,建立慢性脑缺血大鼠模型。假手术组大鼠切开颈部皮肤后分离双侧颈总动脉,但不结扎。使用激光多普勒血流仪检测两组大鼠手术前、手术后(30min内)海马CA1区局部脑血流(r CBF)。术后4周处死大鼠,取出大鼠脑组织并分离海马,采用ELISA法检测IL-1β、IL-6、TNF-α和Aβ1-42表达水平。比较两组大鼠手术前后r CBF与脑组织IL-1β、IL-6、TNF-α、Aβ1-42表达水平;分析大鼠脑组织IL-1β、IL-6、TNF-α表达水平与Aβ1-42表达水平的相关性。结果手术前两组大鼠r CBF比较差异无统计学意义(P>0.05),手术后模型组大鼠r CBF明显低于假手术组(P<0.05)。模型组大鼠手术后r CBF明显低于手术前(P<0.05),而假手术组大鼠手术前后r CBF比较无统计学差异(P>0.05)。模型组大鼠脑组织IL-1β、TNF-α、IL-6等免疫炎性因子表达水平均高于假手术组(均P<0.05),Aβ1-42表达水平亦高于假手术组(均P<0.05)。大鼠脑组织IL-1β、IL-6、TNF-α及Aβ1-42表达水平均呈正相关(r=0.7755、0.5920、0.5536,均P<0.05)。结论慢性脑缺血大鼠脑组织IL-1β、IL-6、TNF-α等免疫炎性因子与Aβ1-42表达水平明显升高,免疫炎性反应或与慢性脑缺血所导致的脑组织Aβ1-42异常沉积有关。
Objective To detect the expression of IL-1β, IL-6, TNF-α and β-amyloid 1-42(Aβ1-42) in the brain tissue of rat model of chronic cerebral ischemia. Methods Sixteen SD rats were randomly divided into the model group(n=8) and the sham operation group(n=8).Chronic cerebral ischemia was induced by bilateral common carotid artery occlusion in model group, while the bilateral common carotid arteries were separated only in sham operation group. The blood flows in CAI region of hippocampus(r CBF) before and 30 min after operation were measured by laser Doppler flowmetry. When the animals were sacrificed 4 weeks later, the levels of IL-1β, IL-6, TNF-α and Aβ1-42 in rat hippocampus were determined by ELISA and compared between two groups. Person correlation analysis between immune/inflammatory factors and Aβ1-42 was performed. Results There was no significant difference in r CBF between the two groups before operation(P >0.05).Postoperative r CBF in the model group was significantly lower than that in the sham operation group. The r CBF in the model group decreased significantly after the operation(P<0.01); while there was no significant difference in r CBF before and after operation in the sham group(P >0.05). The levels of IL-1β, TNF-α, IL-6 and Aβ1-42 in the hippocampus of the model group were higher than those in the sham operation group(P <0.05). And there were significant correlations between the levels of IL-1β and Aβ1-42(r=0.7755, P <0.05), between IL-6 and Aβ1-42( r=0.5920, P<0.05), between TNF-α and Aβ1-42 in the hippocampus(r=0.5536, P<0.05). Conclusion This study indicates that the levels of IL-1β, IL-6, TNF-α and Aβ1-42 in the brain of rat model of chronic cerebral ischemia are significantly increased, which may be associated with immune/inflammatory responses secondary to chronic cerebral ischemia.
Objective To detect the expression of IL-1β, IL-6, TNF-α and β-amyloid 1-42(Aβ1-42) in the brain tissue of rat model of chronic cerebral ischemia. Methods Sixteen SD rats were randomly divided into the model group(n=8) and the sham operation group(n=8).Chronic cerebral ischemia was induced by bilateral common carotid artery occlusion in model group, while the bilateral common carotid arteries were separated only in sham operation group. The blood flows in CAI region of hippocampus(r CBF) before and 30 min after operation were measured by laser Doppler flowmetry. When the animals were sacrificed 4 weeks later, the levels of IL-1β, IL-6, TNF-α and Aβ1-42 in rat hippocampus were determined by ELISA and compared between two groups. Person correlation analysis between immune/inflammatory factors and Aβ1-42 was performed. Results There was no significant difference in r CBF between the two groups before operation(P >0.05).Postoperative r CBF in the model group was significantly lower than that in the sham operation group. The r CBF in the model group decreased significantly after the operation(P<0.01); while there was no significant difference in r CBF before and after operation in the sham group(P >0.05). The levels of IL-1β, TNF-α, IL-6 and Aβ1-42 in the hippocampus of the model group were higher than those in the sham operation group(P <0.05). And there were significant correlations between the levels of IL-1β and Aβ1-42(r=0.7755, P <0.05), between IL-6 and Aβ1-42( r=0.5920, P<0.05), between TNF-α and Aβ1-42 in the hippocampus(r=0.5536, P<0.05). Conclusion This study indicates that the levels of IL-1β, IL-6, TNF-α and Aβ1-42 in the brain of rat model of chronic cerebral ischemia are significantly increased, which may be associated with immune/inflammatory responses secondary to chronic cerebral ischemia.
引文
[1]Castellani RJ,Rolston RK,Smith MA.Alzheimer Disease[J].Disease-a-month:DM,2010,56(9):484-546.DOI:10.1016/j.disamonth.2010.06.001.
[2]董贤慧,柴锡庆.阿尔茨海默病发病机制研究进展[J].中国老年学杂志,2014,29(20):390-394.DOI:10.3969/j.issn.1005-9202.2014.20.145.
[3]李慧源,姜源,孙晓红.阿尔茨海默病炎性反应机制的研究进展[J].中国临床研究,2016,29(1):130-132.DOI:10.13429/j.cnki.cjcr.2016.01.041.
[4]金平,潘永明,潘智勇,等.高胆固醇饮食对兔脑血流量和β-淀粉样蛋白1-42水平的影响[J].浙江医学,2018,40(17):1899-1903.DOI:10.12056/j.issn.1006-2785.2018.40.17.2018-1198.
[5]Jc DLT,Fortin T,Park GA,et al.Chronic cerebrovascular insufficiency induces dementia-like deficits in aged rats[J].Brain Research,1992,582(2):186-195.DOI:10.1016/0006-8993(92)90132-S.
[6]韩德雄,刘喆,冯培培,等.电针不同时间介入对慢性脑缺血大鼠局灶性脑血流、认知功能及炎性损伤的影响[J].中国中西医结合杂志,2017,37(6):721-725.DOI:10.7661/j.cjim.20170405.071.
[7]Dubois MF,H bert R.The incidence of vascular dementia in Canada:a comparison with Europe and East Asia[J].Neuroepidemiology,2001,20(3):179-187.DOI:10.1159/000054785.
[8]Torre JCDL.Alzheimer Disease as a Vascular Disorder Nosological Evidence[J].Stroke;a journal of cerebral circulation,2002,33(9):2147-2148.DOI:10.1161/01.STR.0000014421.15948.67.
[9]杜烨鸿,王凌晞,姚秋会,等.慢性脑缺血对APP/PS1双转基因小鼠行为学及脑组织形态学的影响[J].重庆医科大学学报,2015,40(1):8-12.DOI:10.13406/j.cnki.cyxb.000442.
[10]Liu H,Xing A,Wang X,et al.Regulation ofβ-amyloid level in the brain of rats with cerebrovascular hypoperfusion[J].Neurobiology of Aging,2012,33(4):826-831.DOI:10.1016/j.neurobiolaging.2011.05.027.
[11]Smith C,Anderton BH.Dorothy Russell Memorial Lecture The molecular pathology of Alzheimer's disease:are we any closer to understanding the neurodegenerative process?[J].Neuropathology&Applied Neurobiology,2010,20(4):322-338.DOI:10.1111/j.1365-2990.1994.tb00977.x.
[12]Ballabh P,Braun A,Nedergaard M.The blood-brain barrier:an overview:structure,regulation,and clinical implications.[J].International Journal of Cerebrovascular Diseases,2004,16(1):1-13.DOI:10.1016/j.nbd.2003.12.016.
[13]Pluta R,Ulamekkoziol M,Kocki J,et al.Sporadic Alzheimer's Disease Begins as Episodes of Brain Ischemia and Ischemically Dysregulated Alzheimer's Disease Genes[J].Molecular Neurobiology,2013,48(3):500-515.DOI:10.1007/s12035-013-8439-1.
[14]吴文倩,徐衡,赵守财,等.小胶质细胞在脑缺血中的作用[J].国际脑血管病杂志,2017,25(1):91-94.DOI:10.3760/cma.j.issn.1673-4165.2017.01.012.
[15]张芹,李风娟,张拥波,等.小胶质细胞激活和极化在脑缺血中的作用及其机制[J].国际脑血管病杂志,2014,22(4):307-311.DOI:10.3760/cma.j.issn.1673-4165.2014.04.011.
[16]曹霞飞,让蔚清.炎性反应与阿尔茨海默病[J].中国老年学杂志,2009,29(19):2544-2546.DOI:10.3969/j.issn.1005-9202.2009.19.059.
[17]Birch AM.The contribution of astrocytes to Alzheimer's disease[J].Biochemical Society Transactions,2014,42(5):1316-1320.DOI:10.1042/BST20140171.
[18]雷洪涛,王筠,马淑骅,等.IL-1β、TNF-α与阿尔茨海默病的研究进展[J].中国老年学杂志,2014(24):7115-7117.DOI:10.3969/j.issn.1005-9202.2014.24.137.
[19]Shaftel SS,Kyrkanides S,Olschowka JA,et al.Sustained hippocampal IL-1βoverexpression mediates chronic neuroinflammation and ameliorates Alzheimer plaque pathology[J].Journal of Clinical Investigation,2007,117(6):1595-1604.DOI:10.1172/JCI31450.
[20]王钰,张本恕.炎症因子基因多态性及其功能蛋白与阿尔茨海默病的关系[J].国际老年医学杂志,2008,29(2):68-71.DOI:10.3969/j.issn.1674-7593.2008.02.006.
[21]赵婷婷.阿尔兹海默病的免疫学机制及其治疗进展[J].免疫学杂志,2003(s1):138-142.DOI:10.13431/j.cnki.immunol.j.20030179.
[22]Shepherd CE,Thiel E,Mccann H,et al.Cortical inflammation in Alzheimer disease but not dementia with Lewy bodies[J].Archives of Neurology,2000,57(6):817-822.DOI:10.1001/archneur.57.6.817.
[23]Russell AE,Doll DN,Sarkar SN,et al.TNF-αand Beyond:Rapid Mitochondrial Dysfunction Mediates TNF-α-Induced Neurotoxicity[J].Journal of Clinical&Cellular Immunology,2016,7(6):467-470.DOI:10.4172/2155-9899.1000467.
[24]Laws SM,Perneczky R,Wagenpfeil S,et al.TNF polymorphisms in Alzheimer disease and functional implications on CSF betaamyloid levels[J].Human Mutation,2005,26(1):29-35.DOI:10.1002/humu.20180.
[25]Wu YY,Hsu JL,Wang HC,et al.Alterations of the Neuroinflammatory Markers IL-6 and TRAIL in Alzheimer's Disease[J].Dementia&Geriatric Cognitive Disorders Extra,2015,5(3):424-434.DOI.org/10.1159/000439214.
[26]Zhang Y,Hayes A,Pritchard A,et al.Interleukin-6 promoter polymorphism:risk and pathology of Alzheimer's disease[J].Neuroscience Letters,2004,362(2):99-102.DOI:10.1016/j.neulet.2004.03.008.
[27]Qiu Z,Gruol DL.Interleukin-6,β-amyloid peptide and NMDA interactions in rat cortical neurons[J].Journal of Neuroimmunology,2003,139(1):51-57.DOI:10.1016/S0165-5728(03)00158-9.
[2]董贤慧,柴锡庆.阿尔茨海默病发病机制研究进展[J].中国老年学杂志,2014,29(20):390-394.DOI:10.3969/j.issn.1005-9202.2014.20.145.
[3]李慧源,姜源,孙晓红.阿尔茨海默病炎性反应机制的研究进展[J].中国临床研究,2016,29(1):130-132.DOI:10.13429/j.cnki.cjcr.2016.01.041.
[4]金平,潘永明,潘智勇,等.高胆固醇饮食对兔脑血流量和β-淀粉样蛋白1-42水平的影响[J].浙江医学,2018,40(17):1899-1903.DOI:10.12056/j.issn.1006-2785.2018.40.17.2018-1198.
[5]Jc DLT,Fortin T,Park GA,et al.Chronic cerebrovascular insufficiency induces dementia-like deficits in aged rats[J].Brain Research,1992,582(2):186-195.DOI:10.1016/0006-8993(92)90132-S.
[6]韩德雄,刘喆,冯培培,等.电针不同时间介入对慢性脑缺血大鼠局灶性脑血流、认知功能及炎性损伤的影响[J].中国中西医结合杂志,2017,37(6):721-725.DOI:10.7661/j.cjim.20170405.071.
[7]Dubois MF,H bert R.The incidence of vascular dementia in Canada:a comparison with Europe and East Asia[J].Neuroepidemiology,2001,20(3):179-187.DOI:10.1159/000054785.
[8]Torre JCDL.Alzheimer Disease as a Vascular Disorder Nosological Evidence[J].Stroke;a journal of cerebral circulation,2002,33(9):2147-2148.DOI:10.1161/01.STR.0000014421.15948.67.
[9]杜烨鸿,王凌晞,姚秋会,等.慢性脑缺血对APP/PS1双转基因小鼠行为学及脑组织形态学的影响[J].重庆医科大学学报,2015,40(1):8-12.DOI:10.13406/j.cnki.cyxb.000442.
[10]Liu H,Xing A,Wang X,et al.Regulation ofβ-amyloid level in the brain of rats with cerebrovascular hypoperfusion[J].Neurobiology of Aging,2012,33(4):826-831.DOI:10.1016/j.neurobiolaging.2011.05.027.
[11]Smith C,Anderton BH.Dorothy Russell Memorial Lecture The molecular pathology of Alzheimer's disease:are we any closer to understanding the neurodegenerative process?[J].Neuropathology&Applied Neurobiology,2010,20(4):322-338.DOI:10.1111/j.1365-2990.1994.tb00977.x.
[12]Ballabh P,Braun A,Nedergaard M.The blood-brain barrier:an overview:structure,regulation,and clinical implications.[J].International Journal of Cerebrovascular Diseases,2004,16(1):1-13.DOI:10.1016/j.nbd.2003.12.016.
[13]Pluta R,Ulamekkoziol M,Kocki J,et al.Sporadic Alzheimer's Disease Begins as Episodes of Brain Ischemia and Ischemically Dysregulated Alzheimer's Disease Genes[J].Molecular Neurobiology,2013,48(3):500-515.DOI:10.1007/s12035-013-8439-1.
[14]吴文倩,徐衡,赵守财,等.小胶质细胞在脑缺血中的作用[J].国际脑血管病杂志,2017,25(1):91-94.DOI:10.3760/cma.j.issn.1673-4165.2017.01.012.
[15]张芹,李风娟,张拥波,等.小胶质细胞激活和极化在脑缺血中的作用及其机制[J].国际脑血管病杂志,2014,22(4):307-311.DOI:10.3760/cma.j.issn.1673-4165.2014.04.011.
[16]曹霞飞,让蔚清.炎性反应与阿尔茨海默病[J].中国老年学杂志,2009,29(19):2544-2546.DOI:10.3969/j.issn.1005-9202.2009.19.059.
[17]Birch AM.The contribution of astrocytes to Alzheimer's disease[J].Biochemical Society Transactions,2014,42(5):1316-1320.DOI:10.1042/BST20140171.
[18]雷洪涛,王筠,马淑骅,等.IL-1β、TNF-α与阿尔茨海默病的研究进展[J].中国老年学杂志,2014(24):7115-7117.DOI:10.3969/j.issn.1005-9202.2014.24.137.
[19]Shaftel SS,Kyrkanides S,Olschowka JA,et al.Sustained hippocampal IL-1βoverexpression mediates chronic neuroinflammation and ameliorates Alzheimer plaque pathology[J].Journal of Clinical Investigation,2007,117(6):1595-1604.DOI:10.1172/JCI31450.
[20]王钰,张本恕.炎症因子基因多态性及其功能蛋白与阿尔茨海默病的关系[J].国际老年医学杂志,2008,29(2):68-71.DOI:10.3969/j.issn.1674-7593.2008.02.006.
[21]赵婷婷.阿尔兹海默病的免疫学机制及其治疗进展[J].免疫学杂志,2003(s1):138-142.DOI:10.13431/j.cnki.immunol.j.20030179.
[22]Shepherd CE,Thiel E,Mccann H,et al.Cortical inflammation in Alzheimer disease but not dementia with Lewy bodies[J].Archives of Neurology,2000,57(6):817-822.DOI:10.1001/archneur.57.6.817.
[23]Russell AE,Doll DN,Sarkar SN,et al.TNF-αand Beyond:Rapid Mitochondrial Dysfunction Mediates TNF-α-Induced Neurotoxicity[J].Journal of Clinical&Cellular Immunology,2016,7(6):467-470.DOI:10.4172/2155-9899.1000467.
[24]Laws SM,Perneczky R,Wagenpfeil S,et al.TNF polymorphisms in Alzheimer disease and functional implications on CSF betaamyloid levels[J].Human Mutation,2005,26(1):29-35.DOI:10.1002/humu.20180.
[25]Wu YY,Hsu JL,Wang HC,et al.Alterations of the Neuroinflammatory Markers IL-6 and TRAIL in Alzheimer's Disease[J].Dementia&Geriatric Cognitive Disorders Extra,2015,5(3):424-434.DOI.org/10.1159/000439214.
[26]Zhang Y,Hayes A,Pritchard A,et al.Interleukin-6 promoter polymorphism:risk and pathology of Alzheimer's disease[J].Neuroscience Letters,2004,362(2):99-102.DOI:10.1016/j.neulet.2004.03.008.
[27]Qiu Z,Gruol DL.Interleukin-6,β-amyloid peptide and NMDA interactions in rat cortical neurons[J].Journal of Neuroimmunology,2003,139(1):51-57.DOI:10.1016/S0165-5728(03)00158-9.