【作者】 刘琴；

【导师】 吴成斯；

【作者基本信息】 南昌大学 ， 临床医学硕士（专业学位）， 2025， 硕士

【摘要】 目的:本研究旨在应用simoa技术检测对比阿尔茨海默病(AD)患者及非AD患者血液中磷酸化tau蛋白异构体(p-tau217、p-tau181)和神经丝轻链蛋白(NFL)水平,探讨其在AD早期诊断、鉴别诊断及疾病分期、分型中的价值,并分析其与心理-遗传-结构的相关性。方法:本研究共随机纳入在南昌大学第一附属医院神经内科住院部确诊存在认知障碍受损的受试者(n=42),其中经两名以上神经内科主治医师及以上诊断为阿尔茨海默病组(AD组,n=28),非阿尔茨海默病源性的认知障碍组(非AD组,n=14)。要求入组前必须完善脑脊液痴呆生物标志物检测或痴呆相关PET-CT生物标志物诊断,同时对入组患者完善基线资料收集及评估,完善神经心理量表MMSE、Mo CA、CDR、ADL、HAMA、HAMD、NPI评分等和血液指标检测。通过单分子阵列技术(simoa)测定血液中p-tau181、p-tau217和NFL的水平。将本次检测的血液生物标志物作为阿尔茨海默病的诊断的指标,利用受试者工作曲线(ROC)去分析单个磷酸化tau蛋白或NFL对于诊断阿尔茨海默病的价值;并分析其在轻度认知障碍(MCI)时期与痴呆时期的表达差异,APOE-ε4携带状态、纯合子与杂合子、及典型与不典型AD在上述血液生物标志物水平上的差异性,分析认知评分APOE-ε4携带状态、内侧颞叶萎缩视觉评估量表(Medial Temporal Atrophy,MTA)与生物标志物的相关性。结果:1、血液生物标志物水平比较:(1)AD组患者血液p-tau217及p-tau181水平显著高于非AD组(P<0.0001),但NFL水平在两组间无显著差异。在AD组内,痴呆组(Dementia组)患者的血液p-tau217水平显著高于轻度认知障碍组(MCI组)(P=0.031)。(2)非AD组内,MCI组与Dementia组间NFL、p-tau217、p-tau181均无显著差异(P均>0.05)。典型AD与非典型AD对比,p-tau217也存在显著差异性(P=0.047)。2、神经心理量表评分比较(1)AD组患者MMSE、Mo CA、CDR、ADL评分显著低于非AD组(P均<0.05);(2)AD组和非AD组组内精神症状评分比较,HAMA评分均有显著差异性(P均<0.05)。3、APOEε4与海马相分级分析:AD组中APOEε4携带者比例显著高于非AD组(P=0.047)。在AD组中,且APOE-ε4携带者组中血液p-tau181显著升高(P=0.021)。AD组内,Dementia组患者海马相分级显著高于MCI组(P=0.028)。4、相关性分析:(1)血液生物标志物组合与神经心理量表的相关性分析:1)AD组,(1)NFL与神经心理量表评分均无显著相关性;(2)p-tau181与MMSE、Mo CA、CDR、HAMA、HAMD无显著相关性;(3)p-tau217与MMSE、Mo CA的相关性显著负相关,与CDR之间存在显著正相关,且均具有统计学意义(ρ=﹣0.415、ρ=﹣0.413、ρ=0.405,P值均<0.05)。(4)CDR与HAMA和HAMD之间均存在显著的负相关,具有统计学意义(ρ=﹣0.693、ρ=﹣0.677,P值均<0.05)。2)非AD组,(1)NFL与MMSE、Mo CA呈显著负相关(ρ=﹣0.594、ρ=﹣0.696,P值均<0.05),与CDR呈显著正相关(ρ=0.558,P<0.05);(2)p-tau181、p-tau217与神经心理量表评分之间均无显著相关性;(3)CDR与HAMA显著负相关(ρ=﹣0.634,P<0.05)。(2)血液生物标志物组合与Apo E-ε4基因和MTA评分的相关性分析:(1)p-tau181与Apo E-ε4基因携带呈显著弱相关(ρ=0.361,P=0.019);(2)p-tau217和NFL与Apo E-ε4基因携带无显著相关性。(3)p-tau181和p-tau217与MTA评分呈显著弱相关(ρ=0.359,P=0.020;ρ=0.326,P=0.035)。结论:1、血液p-tau217与p-tau181水平对于诊断阿尔茨海默病有重要作用,而血液p-tau217可作为AD早期诊断和疾病分期的关键生物标志物,且p-tau217可能是区分典型与不典型AD的潜在生物标志物。APOE中ε4携带可能影响血液p-tau181水平。血液NFL可能是非AD源认知损害进展的潜在生物标志物。2、生物-心理-遗传-结构模型有助于AD的早期诊断及预防。更多还原

【Abstract】 Objective:This study aims to apply simoa technology to detect and compare the levels of phosphorylated tau protein isoforms(p-tau217,p-tau181)and neurofilament light chain protein(NFL)in the blood of patients with Alzheimer’s disease(AD)and non-AD individuals.The objective is to explore their value in the early diagnosis,differential diagnosis,disease staging,and typing of AD,as well as to analyze their correlations with psycho-genetic-structural factors.Methods:This study randomly enrolled subjects(n=42)with confirmed cognitive impairment from the Neurology Department of the First Affiliated Hospital of Nanchang University.Among them,subjects diagnosed with Alzheimer’s disease(AD group,n=28)by at least two attending neurologists or above,and subjects with non-Alzheimer’s disease-related cognitive impairment(non-AD group,n=14)were included.Prior to enrollment,all subjects were required to complete cerebrospinal fluid dementia biomarker tests or PET-CT biomarker diagnoses related to dementia.Additionally,baseline data collection and assessments were conducted for all enrolled subjects,including neuropsychological scales such as MMSE,Mo CA,CDR,ADL,HAMA,HAMD,and NPI scores,as well as blood index tests.The levels of p-tau181,p-tau217,and NFL in the blood were measured using Single Molecule Array(Simoa)technology.The blood biomarkers detected in this study were used as diagnostic indicators of Alzheimer’s disease,and the Receiver Operating Characteristic(ROC)curve was employed to analyze the value of individual phosphorylated tau proteins or NFL in the diagnosis of Alzheimer’s disease.Furthermore,the study analyzed the expression differences of these biomarkers between the mild cognitive impairment(MCI)stage and the dementia stage,as well as the differences in blood biomarker levels among APOE-ε4 carriers,homozygotes vs.heterozygotes,and typical vs.atypical AD.The correlation between cognitive scores,APOE-ε4 carrier status,Medial Temporal Atrophy(MTA)visual assessment scale,and biomarkers was also investigated.Results:1、Comparison of Blood Biomarker Levels:(1)P-tau217 and p-tau181 levels were significantly higher in the AD group than in the non-AD group(P<0.0001),while NFL levels showed no significant difference between groups.Within the AD group,p-tau217 levels in the dementia subgroup were significantly higher than those in the MCI subgroup(P=0.031).(2)In the non-AD group,no significant differences in NFL,p-tau217,or p-tau181 levels were observed between the MCI and dementia subgroups(all P>0.05).Additionally,p-tau217 levels differed significantly between typical and atypical AD(P=0.047).2、Neuropsychological Scale Scores:(1)AD patients scored significantly lower than non-AD individuals on the MMSE,Mo CA,CDR,and ADL scales(all P<0.05).(2)Within both the AD and non-AD groups,significant differences in HAMA scores were observed between subgroups(all P<0.05).3、APOEε4 and Hippocampal Phase Grading:AD group had a significantly higher proportion of APOEε4 carriers than the non-AD group(P=0.047).Among AD patients,APOEε4 carriers exhibited elevated plasma p-tau181 levels(P=0.021).Within the AD group,hippocampal phase grading was significantly higher in the dementia subgroup than in the MCI subgroup(P=0.028).4、Correlation Analyses:(1)Blood Biomarkers and Neuropsychological Scales:1)In the AD group:(1)NFL showed no significant correlations with neuropsychological scales.(2)p-tau181 showed no significant correlations with MMSE,Mo CA,CDR,HAMA,or HAMD scores.(3)p-tau217 correlated negatively with MMSE and Mo CA(ρ=-0.415,-0.413)and positively with CDR(ρ=0.405;all P<0.05).(4)CDR correlated negatively with HAMA and HAMD(ρ=-0.693,-0.677;all P<0.05).2)In the non-AD group:(1)NFL correlated negatively with MMSE and Mo CA(ρ=-0.594,-0.696)and positively with CDR(ρ=0.558;all P<0.05).(2)p-tau181 and p-tau217 showed no significant correlations with neuropsychological scales.(3)CDR correlated negatively with HAMA(ρ=-0.634,P<0.05).(2)Blood Biomarkers,APOEε4,and MTA Scores:(1)p-tau181 weakly correlated with APOEε4 carrier status(ρ=0.361,P=0.019)and MTA scores(ρ=0.359,P=0.020).(2)p-tau217 and NFL showed no significant correlations with APOEε4 carrier status.(3)p-tau217 weakly correlated with MTA scores(ρ=0.326,P=0.035).Conclusions:1、Blood p-tau217 and p-tau181 are critical for Alzheimer’s disease(AD)diagnosis.Specifically,p-tau217 may serve as a key biomarker for early AD diagnosis,disease staging,and differentiating typical from atypical AD.APOEε4 carrier status may influence p-tau181 levels.NFL levels may represent a promising biomarker for stratifying disease progression in non-AD etiologies of cognitive impairment.2、A biopsychogenetic-structural model(integrating biomarkers,neuropsychological assessments,APOEε4,and hippocampal grading)could enhance early AD diagnosis and prevention.更多还原