阿尔茨海默病老年患者外周血相关生物学标志物表达及富硒食品的干预效果

doi:10.13481/j.1671-587X.20250520

摘要/Abstract

摘要：

目的检测阿尔茨海默病（AD）患者外周血中AD相关生物学标志物，探讨AD患者血液中抗氧化功能指标活性和水平、相关基因和蛋白表达及富硒饮食干预后的变化情况。方法采用简易智力量表（MMSE）结合脑电图或脑CT及临床医生诊断进行AD筛查。纳入75~90岁老年AD患者56例，其中28例作为AD正常饮食组（AD组），28例作为饮食硒干预组（Se-AD组），同时抽取30名同年龄者作为健康对照组。Se-AD组患者给予日常饮食硒补充（每日增加饮食硒15~20 μg），持续3个月。采用试剂盒和酶联免疫吸附试验（ELISA）法检测各组研究对象血清中超氧化物歧化酶（SOD）、胆碱酯酶（CHE）和谷胱甘肽过氧化物酶（GSH-Px）活性及丙二醛（MDA）、同型半胱氨酸（Hcy）和一氧化氮（NO）水平以及血清中β淀粉样蛋白（Aβ）、微管相关蛋白（Tau）和磷酸化微管相关蛋白（p-Tau）水平，实时荧光定量PCR（RT-qPCR）法检测各组研究对象血液中载脂蛋白E4（ApoE4）、早老素1（PS1）、早老素2（PS2）、半胱氨酸天冬氨酸蛋白酶3（Caspase3）、分拣相关蛋白受体1（SORL1）、β-淀粉样前体蛋白裂解酶1（BACE1）、缺氧诱导因子1（HIF1）、核因子κB（NF-κB）、β淀粉样前体蛋白（APP）、蛋白激酶C（PKC）和Aβ mRNA表达水平。结果与健康对照组比较，Se-AD组和AD组患者血清中SOD活性明显降低（P<0.05），CHE活性和MDA及Hcy水平明显升高（P<0.05）；AD组患者血清中GSH-Px活性明显降低（P<0.05），NO水平明显升高（P<0.05）。与Se-AD组比较，AD组患者血清中CHE活性和Hcy水平明显升高（P<0.05）。Se-AD组和AD组患者血液中ApoE4、PS1、Caspase3、BACE1、NF-κB和APP mRNA表达水平均明显升高（P<0.05），PKC mRNA表达水平均明显降低（P<0.05）；AD组患者血液中PS2 mRNA表达水平明显升高（P<0.05）；Se-AD组和AD组患者血液中Aβ mRNA表达水平明显升高（P<0.05）。结论 AD患者血清中SOD、GSH-Px和CHE 活性，MDA、NO和Hcy水平，Aβ、Tau蛋白和p-Tau蛋白水平以及外周血液中ApoE4、PS1、Caspase3、BACE1、NF-κB、PKC、PS2、Aβ和APP mRNA表达水平有变化，可用于AD患者临床诊断的参考；富硒食品对AD有一定改善作用，其作用机制与减少脑组织氧化损伤及降低AD相关基因PS2和Aβ表达有关。

关键词: 阿尔茨海默病, 富硒食物, β淀粉样蛋白, 同型半胱氨酸, 微管相关蛋白

Abstract:

Objective To detect the biological markers related to Alzheimer’s disease（AD） in the peripheral blood of AD patients， and to explore the activities and levels of the antioxidant function indexes and the expressions of related genes and proteins in the blood of AD patients and the changes after intervention of selenium-rich food. Methods The Mini-Mental State Examination （MMSE） combined with electroencephalogram or brain CT and clinician diagnosis were used for screening AD. Fifty-six elderly patients with AD aged 75-90 years old were selected. Among them，28 cases were selected as normal diet group for AD （AD group）， and 28 cases were selected as dietary selenium intervention group （Se-AD group）. The patients in Se-AD group were given daily dietary selenium supplementation （increaseing dietary selenium by 15-20 μg per day） for 3 months.Meanwhile， 30 people with the same age were selected as healthy control group. The activities of serum superoxide dismutase（SOD）， cholinesterase（CHE）， and glutathione peroxidase （GSH-Px） and the levels of serum malondialdehyde （MDA）， homocysteine （Hcy）， and nitric oxide （NO） as well as reagent kit the levels of serum β-amyloid protein （Aβ）， and microtubule-associated protein （Tau） and phosphorylated microtubule-associated protein（p-Tau） of the subjects in various groups were detected by and enzyme-linked immunosorbent assay（ELISA） method； the expression levels of apolipoprotein E4 （ApoE4）， presenilin 1 （PS1）， presenilin 2 （PS2）， cysteinyl aspartate specific proteinase 3 （Caspase3）， sorting associated protein receptor 1 （SORL1）， β-site amyloid precursor protein cleaving enzyme 1 （BACE1）， hypoxia-inducible factor 1 （HIF1）， nuclear factor-kappa B （NF-κB）， β-amyloid precursor protein （APP）， protein kinase C （PKC）， and Aβ mRNA in peripheral blood of the subjects various groups were detected by real-time fluorescence quantitative PCR （RT-qPCR） method. Results Compared with healthy control group， the serum SOD activities of the patients in Se-AD group and AD group were significantly decreased （P<0.05）， while serum CHE activity and the levels of MDA and Hcy were significantly increased （P<0.05）；the serum GSH-Px activity of the patients in AD group was significantly decreased （P< 0.05）， and the level of NO was significantly increased （P<0.05）. Compared with Se-AD group， serum CHE activity and the level of Hcy of the patients in AD group were significantly increased （P<0.05）. The expression levels of ApoE4， PS1， Caspase3， BACE1， NF-κB and APP mRNA of the patients in Se-AD group and AD group were significantly increased （P<0.05）， and the expression levels of PKC mRNA were significantly decreased （P<0.05）； the expression level of PS2 mRNA of the patients in AD group was significantly increased （P<0.05）， and the expression levels of Aβ mRNA of the patients in Se-AD group and AD group were significantly increased （P<0.05）. Conclusion The activities of serum SOD， GSH-Px and CHE and the levels of MDA， Hcy and NO， the levels of Aβ， Tau and p-Tau proteins，and the expression levels of ApoE4， PS1， Caspase3， BACE1， NF-κB， PKC， PS2， Aβ and APP mRNA in peripheral blood of the AD patients may vary and can be used for clinical diagnosis of the AD patients．Selenium-rich food can improve AD to some extent， and its mechanism is related to reducing the oxidative damage of brain tissue and decreasing the expression of AD related genes PS2 and Aβ．

Key words: Alzheimer’s disease, Selenium-rich food, β-amyloid protein, Homocysteine, Microtubule-associated protein

中图分类号:

R285.5

孙维琦,朱凌羽,徐小磊,刘莹,吕红梅,赖亚辉. 阿尔茨海默病老年患者外周血相关生物学标志物表达及富硒食品的干预效果[J]. 吉林大学学报(医学版), 2025, 51(5): 1333-1339.

Weiqi SUN,Lingyu ZHU,Xiaolei XU,Ying LIU,Hongmei LYU,Yahui LAI. Expressions of peripheral blood related biological markers in elderly patients with Alzheimer’s disease and intervention effect of selenium-rich food[J]. Journal of Jilin University(Medicine Edition), 2025, 51(5): 1333-1339.

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参考文献 25

[1]	TAHAMI MONFARED A A， BYRNES M J， WHITE L A， et al. Alzheimer’s disease： epidemiology and clinical progression［J］. Neurol Ther， 2022， 11（2）： 553-569.
[2]	ZHANG H Q， WEI W， ZHAO M， et al. Interaction between Aβ and Tau in the pathogenesis of Alzheimer’s disease［J］. Int J Biol Sci， 2021， 17（9）： 2181-2192.
[3]	STEFANOSKA K， GAJWANI M， TAN A R P， et al. Alzheimer’s disease： ablating single master site abolishes tau hyperphosphorylation［J］. Sci Adv， 2022， 8（27）： eabl8809.
[4]	ROSTAGNO A A. Pathogenesis of Alzheimer’s disease［J］. Int J Mol Sci， 2022， 24（1）： 107.
[5]	SHI T R， SONG J X， YOU G Y， et al. The function of selenium in central nervous system： lessons from MsrB1 knockout mouse models［J］. Molecules， 2021， 26（5）：1372.
[6]	GHELICHKHANI F， GONZALEZ F A， KAPITONOVA M A， et al. Selenoprotein S： a versatile disordered protein［J］. Arch Biochem Biophys， 2022， 731： 109427.
[7]	YU S S， DU J L. Current views on selenoprotein S in the pathophysiological processes of diabetes-induced atherosclerosis：potential therapeutics and underlying biomarkers［J］. Diabetol Metab Syndr，2024，16（1）：5.
[8]	LI F N， SHI Z， CHENG M N， et al. Biology and roles in diseases of selenoprotein I characterized by ethanolamine phosphotransferase activity and antioxidant potential［J］. J Nutr， 2023， 153（11）： 3164-3172.
[9]	MASUDA　R， KUWANO S， GOTO K. Modeling selenoprotein Se-nitrosation： synthesis of a Se-nitrososelenocysteine with persistent stability［J］. J Am Chem Soc， 2023， 145（26）： 14184-14189.
[10]	胡承平，陶凤芝，王灿，等. 星形胶质细胞在阿尔茨海默病中的研究进展［J］. 同济大学学报（医学版）， 2023，44（05）： 740-746.
[11]	SHERIFF S， SHEN T， ABDAL S，et al. Retinal thickness and vascular parameters using optical coherence tomography in Alzheimer’s disease：a meta-analysis［J］. Neural Regen Res，2023，18（11）：2504-2513.
[12]	VOGEL J W， ITURRIA-MEDINA Y， STRANDBERG O T， et al. Spread of pathological tau proteins through communicating neurons in human Alzheimer’s disease［J］. Nat Commun， 2020， 11（1）： 2612.
[13]	KAC P R， GONZALEZ-ORTIZ F， SIMRÉN J， et al. Diagnostic value of serum versus plasma phospho-tau for Alzheimer’s disease［J］. Alzheimers Res Ther， 2022， 14（1）： 65.
[14]	YUE Q， HOI M P M. Emerging roles of astrocytes in blood-brain barrier disruption upon amyloid-beta insults in Alzheimer’s disease［J］. Neural Regen Res， 2023， 18（9）： 1890-1902.
[15]	FENG W X， ZHANG Y L， SUN P， et al. Acquired immunity and Alzheimer’s disease［J］. J Biomed Res， 2022， 37（1）： 15-29.
[16]	CHEN Z Y， ZHANG Y. Animal models of Alzheimer’s disease： Applications， evaluation，and perspectives［J］. Zool Res， 2022， 43（6）： 1026-1040.
[17]	SU L N， LI R Q， ZHANG Z Q， et al. Identification of altered exosomal microRNAs and mRNAs in Alzheimer’s disease［J］. Ageing Res Rev， 2022， 73：101497.
[18]	BOTTO R， CALLAI N， CERMELLI A， et al. Anxiety and depression in Alzheimer’s disease： a systematic review of pathogenetic mechanisms and relation to cognitive decline［J］. Neurol Sci， 2022， 43（7）：4107-4124.
[19]	CIOFFI F， ADAM R H I， BANSAL R， et al. A review of oxidative stress products and related genes in early Alzheimer’s disease［J］. J Alzheimers Dis， 2021， 83（3）： 977-1001.
[20]	WU Y G， SONG L J， YIN L J， et al. The effects and potential of microglial polarization and crosstalk with other cells of the central nervous system in the treatment of Alzheimer’s disease［J］. Neural Regen Res， 2023， 18（5）： 947-954.
[21]	HUANG C W， RUST N C， WU H F， et al. Altered O-GlcNAcylation and mitochondrial dysfunction， a molecular link between brain glucose dysregulation and sporadic Alzheimer’s disease［J］. Neural Regen Res， 2023， 18（4）： 779-783.
[22]	YEATES C， DESHPANDE P， KANGO-SINGH M， et al. Signaling interactions among neurons impact cell fitness and death in Alzheimer’s disease［J］. Neural Regen Res， 2023， 18（4）： 784-789.
[23]	SI Z Z， ZOU C J， MEI X， et al. Targeting neuroinflammation in Alzheimer’s disease： from mechanisms to clinical applications［J］. Neural Regen Res， 2023， 18（4）： 708-715.
[24]	MAISAM M， KHAN M T， LODHI M S， et al. Alzheimer’s disease： mechanism， mutations， and applications of nano-medicine［J］. Front Biosci， 2023， 28（10）： 258.
[25]	GHOLAMI A. Alzheimer’s disease： The role of proteins in formation， mechanisms， and new therapeutic approaches［J］. Neurosci Lett， 2023， 817： 137532.

Primer	Sequence	Fragment length （bp）
ApoE4	F：5'-CTGCGTTGCTGGTCACATTC-3'	146
	R：5'-GTAATCCCAAAAGCGACCCAG-3'
PS1	F：5'-TTACCTGCACCGTTGTCCTACTT-3'	188
	R：5'-TCTTCTTCCTCATCTTGCTCCAC-3'
PS2	F：5'-GAGCAGAGAGGAGGCAATGG-3'	191
	R：5'-CGAAACAGCAGCCCTTATTTG-3'
Caspase3	F：5'-TGGAAGCGAATCAATGGACTCT-3'	170
	R：5'-TGGAAGCGAATCAATGGACTCT-3'
SORL1	F：5'-TCTACATTGAACGACATGAACCCTC-3'	298
	R：5'-ACACACACAAACACCTGGTCCT-3'
BACE1	F：5'-TCATTGTGCGGGTGGAGAT-3'	149
	R：5'-GGCTGCCTTGATGGATTTG-3'
HIF1	F：5'-TGATTGCATCTCCATCTCCTACC-3'	177
	R：5'-GACTCAAAGCGACAGATAACACG-3'
NF-κB	F：5'-GACGCATTGCTGTGCCTTC-3'	91
	R：5'-TTGATGGTGCTCAGGGATGAC-3'
APP	F：5'-TGCAATCATTGGACTCATGG-3'	164
	R：5'-TCTGCTGCATCTTGGACAGG-3'
PKC	F：5'-TGACTACATCGCCCCAGAGA-3'	82
	R：5'-ATAAAGGAGAACCCCGAAGGA-3'
Aβ	F：5'-GGGTTCAAACAAAGGTGCAATC-3'	198
	R：5'-GTTGGATTTTCGTAGCCGTTCT-3'