Background Neuropathologically, Alzheimer disease （AD） is characterized by the presence of extracellular plaques enriched in β-amyloid peptides; however, the mechanism by which it results in the neurotoxicity is un...Background Neuropathologically, Alzheimer disease （AD） is characterized by the presence of extracellular plaques enriched in β-amyloid peptides; however, the mechanism by which it results in the neurotoxicity is uncertain. The purpose of this study was to investigate whether it would prompt the progress of Alzheimer disease via enhancement of aberrant phosphorylated tau that results from its increased kinase gene expression. Methods Twenty-four male rats were divided into three groups, and each group had 8 rats： control, sham-operated, and Aβ25-35 injected AD model groups. AD rat models were created by unilateral injections of Aβ25-35 into the amygdala. The hyperphosphorylated tau protein was estimated by immunohistochemistry with paired helical filament-1 （PHF-1） antibody and paired helical filament-tau （AT8） antibody. The expressions of glycogen synthase kinase-3β （GSK-3β） and p38 mitogen-activated protein kinase （P38MAPK） mRNA were observed by in situ hybridization. Results Compared with the control and sham-operated groups, the evaluation of paired AT8 and paired helical filament-1 （PHF-1） in the cortexes and hippocampus of the AD model group showed the numbers of AT8 and PHF-1 positive cells, as well as the optical density （OD） values of the proteins were significantly higher （AT8： in CA2： 0.318±0.037 vs. 0.135±0.028, 0.136±0.031; in frontal cortex： 0.278±0.040 vs. 0.130±0.028, 0.190±0.037. PHF-1 ： in CA2： 0.386±0.034 vs. 0.139±0.010, 0.193±0.041; in frontal cortex： 0.395±0.050 vs. 0.159±0.030, 0.190±0.044, respectively, P 〈0.01）; the number of GSK-3β mRNA and P38MAPK mRNA positive cells of the AD model group, as well as the OD values, also increased significantly in the cortexes, hippocampus （GSK-3β-mRNA： in CA2：0.384±0.012 vs. 0.190±0.015, 0.258±0.064; in frontal cortex： 0.398±0.018 vs. 0.184±0.031, 0.218±0.049. P38MAPK mRNA： in CA2：0.409±0.038 vs. 0.161±0.041, 0.189±0.035; in frontal cortex： 0.423±0.070 vs. 0.160±0展开更多
目的寻找基于蛋白质组学技术早期、快速诊断糖尿病肾病的尿蛋白标志物模型，并探讨其临床应用价值。方法应用表面增强激光解析电离飞行时间质谱（surface-enhanced laser desorption-ionization time of flight mass spectrometry，SELD...目的寻找基于蛋白质组学技术早期、快速诊断糖尿病肾病的尿蛋白标志物模型，并探讨其临床应用价值。方法应用表面增强激光解析电离飞行时间质谱（surface-enhanced laser desorption-ionization time of flight mass spectrometry，SELDI-TOF-MS）技术及Au芯片（proteinchipgoldarray）检测292例患者尿蛋白质谱，包括129例糖尿病肾病和163例对照者（61例糖尿病及102名健康体检者）。获得的蛋白质谱数据用Biomaker Wizard3．1软件筛选差异蛋白，通过生物标志模型软件（biomarker patterns software，BPS）建立决策树辨别分析模型，评价其临床诊断价值。对部分筛选的差异蛋白通过比对标准蛋白质谱数据，根据分子量大小进行初步鉴定。结果糖尿病肾病患者与对照者尿液中差异表达的蛋白质峰有40个，其丰度值两组间比较差异均有统计学意义（t值为-9．81～24．52，P均〈0．05），通过BPS自动筛选66916质荷比（m／z）蛋白建立的模型诊断糖尿病肾病敏感度为98．7％（78／79），特异度98．2％（111／113）。对糖尿病和糖尿病肾病患者尿蛋白质谱图分析后得到24个差异蛋白质峰，其丰度值两组间比较差异均有统计学意义（t值为-6．95～14．45，P均〈0．05），BPS筛选4008、11619、66916m／z蛋白建立模型区分糖尿病与糖尿病肾病的敏感度（129／129）和特异度（61／61）均为100％。通过比对标准蛋白质谱数据，糖尿病肾病患者尿差异蛋白中m／z 11619、23529、66916和79378，可能为β2-微球蛋白、α1-微球蛋白、白蛋白和转铁蛋白。结论基于SELDI-TOF-MS及Au芯片技术检测尿蛋白质谱在鉴别蛋白尿来源、糖尿病肾病的早期快速诊断及肾脏损害评估具有重要应用价值。展开更多
基金This work was supported by the National Natural Science Foundation of China (No.30271611) the Key Science and Technology Research Project of Fujian Province (No.2003F009).
基金This study was supported by the grants from the National Natural Science Foundation of China （No. 30971029）, Fund of Shanghai Science and Technology Committee （No. 08411950900）, and Young Excellent Talents Award of Tongji University （No. 2006KJ067）.
文摘Background Neuropathologically, Alzheimer disease （AD） is characterized by the presence of extracellular plaques enriched in β-amyloid peptides; however, the mechanism by which it results in the neurotoxicity is uncertain. The purpose of this study was to investigate whether it would prompt the progress of Alzheimer disease via enhancement of aberrant phosphorylated tau that results from its increased kinase gene expression. Methods Twenty-four male rats were divided into three groups, and each group had 8 rats： control, sham-operated, and Aβ25-35 injected AD model groups. AD rat models were created by unilateral injections of Aβ25-35 into the amygdala. The hyperphosphorylated tau protein was estimated by immunohistochemistry with paired helical filament-1 （PHF-1） antibody and paired helical filament-tau （AT8） antibody. The expressions of glycogen synthase kinase-3β （GSK-3β） and p38 mitogen-activated protein kinase （P38MAPK） mRNA were observed by in situ hybridization. Results Compared with the control and sham-operated groups, the evaluation of paired AT8 and paired helical filament-1 （PHF-1） in the cortexes and hippocampus of the AD model group showed the numbers of AT8 and PHF-1 positive cells, as well as the optical density （OD） values of the proteins were significantly higher （AT8： in CA2： 0.318±0.037 vs. 0.135±0.028, 0.136±0.031; in frontal cortex： 0.278±0.040 vs. 0.130±0.028, 0.190±0.037. PHF-1 ： in CA2： 0.386±0.034 vs. 0.139±0.010, 0.193±0.041; in frontal cortex： 0.395±0.050 vs. 0.159±0.030, 0.190±0.044, respectively, P 〈0.01）; the number of GSK-3β mRNA and P38MAPK mRNA positive cells of the AD model group, as well as the OD values, also increased significantly in the cortexes, hippocampus （GSK-3β-mRNA： in CA2：0.384±0.012 vs. 0.190±0.015, 0.258±0.064; in frontal cortex： 0.398±0.018 vs. 0.184±0.031, 0.218±0.049. P38MAPK mRNA： in CA2：0.409±0.038 vs. 0.161±0.041, 0.189±0.035; in frontal cortex： 0.423±0.070 vs. 0.160±0
文摘目的寻找基于蛋白质组学技术早期、快速诊断糖尿病肾病的尿蛋白标志物模型，并探讨其临床应用价值。方法应用表面增强激光解析电离飞行时间质谱（surface-enhanced laser desorption-ionization time of flight mass spectrometry，SELDI-TOF-MS）技术及Au芯片（proteinchipgoldarray）检测292例患者尿蛋白质谱，包括129例糖尿病肾病和163例对照者（61例糖尿病及102名健康体检者）。获得的蛋白质谱数据用Biomaker Wizard3．1软件筛选差异蛋白，通过生物标志模型软件（biomarker patterns software，BPS）建立决策树辨别分析模型，评价其临床诊断价值。对部分筛选的差异蛋白通过比对标准蛋白质谱数据，根据分子量大小进行初步鉴定。结果糖尿病肾病患者与对照者尿液中差异表达的蛋白质峰有40个，其丰度值两组间比较差异均有统计学意义（t值为-9．81～24．52，P均〈0．05），通过BPS自动筛选66916质荷比（m／z）蛋白建立的模型诊断糖尿病肾病敏感度为98．7％（78／79），特异度98．2％（111／113）。对糖尿病和糖尿病肾病患者尿蛋白质谱图分析后得到24个差异蛋白质峰，其丰度值两组间比较差异均有统计学意义（t值为-6．95～14．45，P均〈0．05），BPS筛选4008、11619、66916m／z蛋白建立模型区分糖尿病与糖尿病肾病的敏感度（129／129）和特异度（61／61）均为100％。通过比对标准蛋白质谱数据，糖尿病肾病患者尿差异蛋白中m／z 11619、23529、66916和79378，可能为β2-微球蛋白、α1-微球蛋白、白蛋白和转铁蛋白。结论基于SELDI-TOF-MS及Au芯片技术检测尿蛋白质谱在鉴别蛋白尿来源、糖尿病肾病的早期快速诊断及肾脏损害评估具有重要应用价值。
