细胞外基质金属蛋白酶诱导因子与阿尔茨海默病γ-分泌酶的相关性研究
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摘要
目的探讨细胞外基质金属蛋白酶诱导因子(CD147)与阿尔茨海默病γ-分泌酶的相关性。方法 CD147 c DNA转染人神经母细胞瘤,并通过遗传霉素G418筛选CD147超表达细胞。超表达细胞设为实验组,正常细胞设为对照组,Western Blot检测两组CD147含量,Elisa分析两组Aβ40和Aβ42含量。结果实验组和对照组CD147含量分别为(40.23±0.20)μg/μL和(27.98±0.12)μg/μL,实验组显著高于对照组(t=90.97,P<0.05)。实验组和对照组Aβ40与Aβ42总含量分别为(141.05±2.13)pg/m L和(323.41±2.78)pg/m L,实验组显著低于对照组(t=90.19,P<0.05)。结论随着CD147含量的增加,Aβ的生成减少,而γ-分泌酶是生成Aβ的限速酶,可知γ-分泌酶随着CD147含量的增加而呈负增长,提示CD147对γ-分泌酶的活性起着负调节作用。
Objective To investigate the relationship between extracellular matrix metalloproteinase inducer( CD147) and γ-secretase in Alzheimer's disease. Methods CD147 c DNA was transfected into human neuroblastoma cells,and cells with overexpression of CD147 were selected by neomycin G418. CD147 overexpression cells and normal cells were set as experimental group and control group,respectively. The expression of CD147 protein in the two groups were determined by Western blot,and the content of amyloid βprotein( Aβ) 40 and Aβ42 in the two groups was analyzed by enzyme-linked immunosorbent assay. Results The expression of CD147 protein was significantly higher in the experimental group than in the control group( 40. 23 ± 0. 20 vs. 27. 98 ± 0. 12 μg / μl; t =90. 97,P < 0. 05). Moreover,the total content of Aβ40 and Aβ42 was significantly lower in the experimental group than in the control group( 141. 05 ± 2. 13 vs. 323. 41 ± 2. 78 pg / ml; t = 90. 19,P < 0. 05). Conclusions The synthesis of Aβ is reduced with increasing CD147 content. Given that γ-secretase is the rate-limiting enzyme in Aβ synthesis,the level of γ-secretase should be reduced with increasing CD147 content,indicating that CD147 is a negative regulator of γ-secretase activity.
Objective To investigate the relationship between extracellular matrix metalloproteinase inducer( CD147) and γ-secretase in Alzheimer's disease. Methods CD147 c DNA was transfected into human neuroblastoma cells,and cells with overexpression of CD147 were selected by neomycin G418. CD147 overexpression cells and normal cells were set as experimental group and control group,respectively. The expression of CD147 protein in the two groups were determined by Western blot,and the content of amyloid βprotein( Aβ) 40 and Aβ42 in the two groups was analyzed by enzyme-linked immunosorbent assay. Results The expression of CD147 protein was significantly higher in the experimental group than in the control group( 40. 23 ± 0. 20 vs. 27. 98 ± 0. 12 μg / μl; t =90. 97,P < 0. 05). Moreover,the total content of Aβ40 and Aβ42 was significantly lower in the experimental group than in the control group( 141. 05 ± 2. 13 vs. 323. 41 ± 2. 78 pg / ml; t = 90. 19,P < 0. 05). Conclusions The synthesis of Aβ is reduced with increasing CD147 content. Given that γ-secretase is the rate-limiting enzyme in Aβ synthesis,the level of γ-secretase should be reduced with increasing CD147 content,indicating that CD147 is a negative regulator of γ-secretase activity.
引文
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[2]李兴强.家族性阿尔茨海默病研究现状.国际神经病学神经外科杂志,2014,41(2):156-159.
[3]Lippa CF.Alzheimer’s Disease:A Financial Crisis and the Commonwealth of Pennsylvania.Am J Alzheimers Dis Other Demen,2015,30(2):117-118.
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[6]胡海燕.β-淀粉样蛋白在阿尔茨海默病中的作用及中药多靶点对抗研究进展.中华中医药学刊,2012,30(3):488-492.
[7]康林,杜鹃,王斌,等.细胞外基质金属蛋白酶诱导因子降解途径的初步研究.神经解剖学杂志,2014,30(5):541-545.
[8]Kanyenda LJ,Verdile G,Martins R,et al.Is Cholesterol and Amyloid-βStress Induced CD 147Expression a Protective Response?Evidence that Extracellular Cyclophilin A Mediated Neuroprotection is Reliant on CD 147.J Alzheimer’s Dis,2014,39:545-556.
[9]Yurt Kilcar A,Biber Müftüler ZF,Evren V,et al.Investigation of 99m Tc Labeled Clioquinol Derivative on Amyloid Plaque Specifity by Using Animal Model of Alzheimer’s Disease.Mol Imaging Radionucl Ther,2015,24(1):40-41.
[10]Wang L,Benzinger TL,Hassenstab J,et al.Spatially distinct atrophy is linked toβ-amyloid and tau in preclinical Alzheimer disease.Neurology,2015,84(12):1254-1260.
[11]Brickman AM,Guzman VA,Gonzalez-Castellon M,et al.Cerebral autoregulation,beta amyloid,and white matter hyperintensities are interrelated.Neurosci Lett,2015,592:54-58.
[12]Nahalkova J,Volkmann I,Aoki M,et al.CD147,aγ-secretase associated protein is upregulated in Alzheimer’s disease brain and its cellular trafficking is affected by presenilin-2.Neurochem Int,2010,56:67-76.
[13]Roher AE,Maarouf CL,Kokjohn TA,et al.Neuropathological and biochemical assessments of an Alzheimer’s disease patient treated with theγ-secretase inhibitor semagacestat.Am J Neurodegener Dis,2014,3(3):115-133.
[14]Zhou SX,Zhou H,Walian PJ,et al.CD147is a regulatory subunit of theγ-secretase complex in Alzheimer’s disease amyloidβ-peptide production.Proc Natl Acad Sci U S A,2005,102(21):7499-7504.
[15]张锦巍,井丽娟,孙亚平,等.CD147超表达对γ-分泌酶影响的临床研究.重庆医学,2012,41(36):3801-3806.
[2]李兴强.家族性阿尔茨海默病研究现状.国际神经病学神经外科杂志,2014,41(2):156-159.
[3]Lippa CF.Alzheimer’s Disease:A Financial Crisis and the Commonwealth of Pennsylvania.Am J Alzheimers Dis Other Demen,2015,30(2):117-118.
[4]徐金静,方力群,赵春慧,等.β淀粉样蛋白对阿尔茨海默病转基因果蝇认知功能的影响.国际神经病学神经外科杂志,2013,40(2):101-104.
[5]高玲焕,邹莉波,迟天燕.β-淀粉样蛋白在阿尔茨海默病病因学中作用研究进展.沈阳药科大学学报,2008,25:41-45.
[6]胡海燕.β-淀粉样蛋白在阿尔茨海默病中的作用及中药多靶点对抗研究进展.中华中医药学刊,2012,30(3):488-492.
[7]康林,杜鹃,王斌,等.细胞外基质金属蛋白酶诱导因子降解途径的初步研究.神经解剖学杂志,2014,30(5):541-545.
[8]Kanyenda LJ,Verdile G,Martins R,et al.Is Cholesterol and Amyloid-βStress Induced CD 147Expression a Protective Response?Evidence that Extracellular Cyclophilin A Mediated Neuroprotection is Reliant on CD 147.J Alzheimer’s Dis,2014,39:545-556.
[9]Yurt Kilcar A,Biber Müftüler ZF,Evren V,et al.Investigation of 99m Tc Labeled Clioquinol Derivative on Amyloid Plaque Specifity by Using Animal Model of Alzheimer’s Disease.Mol Imaging Radionucl Ther,2015,24(1):40-41.
[10]Wang L,Benzinger TL,Hassenstab J,et al.Spatially distinct atrophy is linked toβ-amyloid and tau in preclinical Alzheimer disease.Neurology,2015,84(12):1254-1260.
[11]Brickman AM,Guzman VA,Gonzalez-Castellon M,et al.Cerebral autoregulation,beta amyloid,and white matter hyperintensities are interrelated.Neurosci Lett,2015,592:54-58.
[12]Nahalkova J,Volkmann I,Aoki M,et al.CD147,aγ-secretase associated protein is upregulated in Alzheimer’s disease brain and its cellular trafficking is affected by presenilin-2.Neurochem Int,2010,56:67-76.
[13]Roher AE,Maarouf CL,Kokjohn TA,et al.Neuropathological and biochemical assessments of an Alzheimer’s disease patient treated with theγ-secretase inhibitor semagacestat.Am J Neurodegener Dis,2014,3(3):115-133.
[14]Zhou SX,Zhou H,Walian PJ,et al.CD147is a regulatory subunit of theγ-secretase complex in Alzheimer’s disease amyloidβ-peptide production.Proc Natl Acad Sci U S A,2005,102(21):7499-7504.
[15]张锦巍,井丽娟,孙亚平,等.CD147超表达对γ-分泌酶影响的临床研究.重庆医学,2012,41(36):3801-3806.