老年性痴呆患者血清miR-375、SP1水平与认知功能的相关性-东南大学学报医学版

老年性痴呆患者血清miR-375、SP1水平与认知功能的相关性

单位：北京航天总医院老年医学科, 北京丰台 100076

分类号：R749.6

出版年·卷·期（页码）：2023·42·第二期（257-266）

摘要：

目的:检测老年性痴呆患者血清微小RNA-375(miR-375)、特异性蛋白1(SP1)表达水平,探讨其与认知功能的相关性。方法:选取2021年4月至2022年4月在本院就诊的老年性痴呆患者120例纳入疾病组,根据临床痴呆评定量表(CDR)评定患者病情严重程度,选取同期在本院进行健康体检的志愿者120例纳入对照组。采用实时荧光定量PCR(qRT-PCR)法检测所有受试者血清miR-375、SP1 mRNA表达水平。采用简易精神状态量表(MMSE)评估所有受试者的认知功能。比较不同病情严重程度患者血清miR-375、SP1 mRNA表达水平及MMSE评分。采用Pearson相关分析疾病组血清miR-375与SP1 mRNA表达水平的相关性,Spearman相关分析两者与MMSE评分的相关性。采用多因素Logistic回归分析影响老年性痴呆发生的危险因素。采用受试者工作特征(ROC)曲线分析血清miR-375、SP1 mRNA表达水平和MMSE评分对老年性痴呆患者病情进展为重度的预测价值。采用双荧光素酶报告基因实验检测miR-375与SP1的关系。体外培养人神经母细胞瘤细胞SH-SY5Y,通过β-淀粉样蛋白肽25-35(Aβ25-35)诱导构建老年性痴呆体外细胞模型,观察过表达miR-375对Aβ25-35诱导的SH-SY5Y细胞中SP1 mRNA和蛋白表达以及细胞活力、凋亡率的影响。结果:疾病组血清miR-375表达水平、MMSE评分显著低于对照组,SP1 mRNA表达水平显著高于对照组(P＜0.05)。重度组、中度组血清miR-375表达水平、MMSE评分显著低于轻度组,SP1 mRNA表达水平显著高于轻度组(P＜0.05);重度组血清miR-375表达水平、MMSE评分显著低于中度组,SP1 mRNA表达水平显著高于中度组(P＜0.05)。Pearson相关分析显示,疾病组血清miR-375与SP1 mRNA表达水平呈负相关(r=-0.627,P＜0.05);Spearman相关分析显示,血清miR-375表达水平与MMSE评分呈正相关(r=0.664,P＜0.05),血清SP1 mRNA表达水平与MMSE评分呈负相关(r=-0.472,P＜0.05)。多因素Logistic回归分析显示,低MMSE评分、miR-375低表达、SP1 mRNA高表达是影响老年性痴呆发生的危险因素(P＜0.05)。ROC曲线分析显示,血清miR-375、SP1 mRNA表达水平和MMSE评分以及三者联合预测老年性痴呆患者病情进展为重度的曲线下面积(AUC)分别为0.823、0.780、0.851、0.938,且三者联合预测的AUC显著高于单个指标预测的AUC(Z=2.734、3.150、2.113,P＜0.05)。双荧光素酶报告基因实验显示miR-375能够靶向SP1。Aβ25-35诱导后,SH-SY5Y细胞中miR-375表达水平、细胞活力显著降低,SP1 mRNA和蛋白表达以及凋亡率显著升高(P＜0.05);过表达miR-375可显著升高Aβ25-35诱导的SH-SY5Y细胞活力,降低SP1 mRNA和蛋白表达以及凋亡率(P＜0.05)。结论:老年性痴呆患者血清miR-375表达水平下调,SP1 mRNA表达水平上调,且均与认知功能密切相关,可能两者通过相互作用参与调控患者病情进展。

Objective: To detect the expression levels of serum microRNA-375(miR-375) and specific protein 1(SP1) in patients with senile dementia, and explore their correlation with cognitive function. Methods: 120 patients with senile dementia who were treated in our hospital from April 2021 to April 2022 were selected as the disease group, and the severity of the disease was evaluated according to the Clinical Dementia Rating Scale(CDR). In addition, 120 volunteers who underwent physical examination in our hospital at the same time were regarded as the control group. Serum miR-375 and SP1 mRNA expression levels in all subjects were detected by real-time quantitative PCR(qRT-PCR). Cognitive function in all subjects was assessed by Mini-Mental State Examination(MMSE). Serum miR-375 and SP1 mRNA expression levels and MMSE scores of patients with different disease severity were compared. Correlation between serum miR-375 and SP1 mRNA expression levels in the disease group was analyzed by Pearson correlation, and correlation between the two and the MMSE score was analyzed by Spearman correlation. Risk factors affecting the occurrence of senile dementia were analyzed by multivariate Logistic regression. Predictive value of serum miR-375, SP1 mRNA expression level and MMSE score on disease progressing to severe in patients with senile dementia was analyzed by receiver operating characteristic(ROC) curve. Relationship between miR-375 and SP1 was analyzed by dual-luciferase reporter gene assay. Human neuroblastoma cell SH-SY5Y was cultured in vitro and senile dementia cell model was constructed by β-amyloid peptide 25-35(Aβ25-35) induction, and the effects of overexpression of miR-375 on the expression of SP1 mRNA and protein, cell viability and apoptosis rate in SH-SY5Y cells induced by Aβ25-35 were observed. Results: Serum miR-375 expression level and MMSE score in the disease group were significantly lower than those in the control group, and SP1 mRNA expression level was significantly higher than that in the control group(P<0.05). Serum miR-375 expression level and MMSE score in the severe and moderate groups were significantly lower than those in the mild group, and SP1 mRNA expression level was significantly higher than that in the mild group(P<0.05); serum miR-375 expression level and MMSE score in the severe group were significantly lower than those in the moderate group, and SP1 mRNA expression level was significantly higher than that in the moderate group(P<0.05). Pearson correlation analysis showed that serum miR-375 expression level was significantly negatively correlated with SP1 mRNA expression level in the disease group(r=-0.627, P<0.05). Spearman correlation analysis showed that serum miR-375 expression level was positively correlated with MMSE score(r=0.664, P<0.05), and serum SP1 mRNA expression level was negatively correlated with MMSE score(r=-0.472, P<0.05). Multivariate Logistic regression analysis showed that low MMSE score, low expression of miR-375 and high expression of SP1 mRNA were risk factors affecting the occurrence of senile dementia(P<0.05). ROC curve analysis showed that area under the curve(AUC) of serum miR-375, SP1 mRNA expression level and MMSE score, and combination of the three for predicting disease progressing to severe in patients with senile dementia were 0.823, 0.780, 0.851 and 0.938, respectively, and the AUC predicted by combination of the three was significantly higher than that predicted by a single index(Z=2.734, 3.150, 2.113, P<0.05). Dual-luciferase reporter gene assay showed that miR-375 could target SP1. After Aβ25-35 induction, miR-375 expression level and cell viability in SH-SY5Y cells were significantly decreased, while SP1 mRNA and protein expression level and apoptosis rate were significantly increased(P<0.05); overexpression of miR-375 can significantly increase cell viability in SH-SY5Y cells induced by Aβ25-35, and decreased SP1 mRNA and protein expression level and apoptosis rate(P<0.05). Conclusion: Serum miR-375 expression level is down-regulated and SP1 mRNA expression level is up-regulated in patients with senile dementia, both of them are closely related to cognitive function, and may participate in the regulation of patients' disease progression through interaction.

参考文献：

[1] JIA L,QUAN M, FU Y,et al.Dementia in China:epidemiology,clinical management,and research advances[J].Lancet Neurol,2020,19(1):81-92.
[2] CHEN F Z,ZHAO Y,CHEN H Z.MicroRNA-98 reduces amyloid β-protein production and improves oxidative stress and mitochondrial dysfunction through the Notch signaling pathway via HEY2 in Alzheimer's disease mice[J].Int J Mol Med,2019,43(1):91-102.
[3] GUGLIANDOLO A,CHIRICOSTA L,BOCCARDI V,et al.MicroRNAs modulate the pathogenesis of Alzheimer's disease:an in Silico analysis in the human brain[J].Genes(Basel),2020,11(9):983.
[4] YANG W B,HSU C C,HSU T I,et al.Increased activation of HDAC1/2/6 and Sp1 underlies therapeutic resistance and tumor growth in glioblastoma[J].Neuro Oncol,2020,22(10):1439-1451.
[5] CITRON B A,SAYKALLY J N,CAO C,et al.Transcription factor Sp1 inhibition,memory,and cytokines in a mouse model of Alzheimer's disease[J].Am J Neurodegener Dis,2015,4(2):40-48.
[6] TURK A,KUNEJ T,PETERLIN B.MicroRNA-target interaction regulatory network in Alzheimer's disease[J].J Pers Med,2021,11(12):1275-1292.
[7] 中国痴呆与认知障碍写作组,中国医师协会神经内科医师分会认知障碍疾病专业委员会.2018中国痴呆与认知障碍诊治指南(二):阿尔茨海默病诊治指南[J].中华医学杂志,2018,98(13):971-977.
[8] 中国老年医学学会认知障碍分会.临床痴呆评定量表简体中文版[J].中华老年医学杂志,2018,37(4):367-371.
[9] GALEA M,WOODWARD M.Mini-mental state examination(MMSE)[J].Aust J Physiother,2005,51(3):198.
[10] 刘洁,米亚静,张典,等.circ_0000285靶向miR-127-5p调控阿尔茨海默病细胞模型的损伤[J].中国免疫学杂志,2022,38(11):1288-1293.
[11] 魏玮,谢晓琳,李学盈,等.老年痴呆患者炎性因子、血糖、血压与认知功能的相关性[J].中国老年学杂志,2020,40(24):5248-5250.
[12] WEI W,WANG Z Y,MA L N,et al.MicroRNAs in Alzheimer's disease:function and potential applications as diagnostic biomarkers[J].Front Mol Neurosci,2020,13:160-175.
[13] DUBOIS B,VILLAIN N,FRISONI G B,et al.Clinical diagnosis of Alzheimer's disease:recommendations of the International Working Group[J].Lancet Neurol,2021,20(6):484-496.
[14] SUN C,LIU J,DUAN F,et al.The role of the microRNA regulatory network in Alzheimer's disease:a bioinformatics analysis[J].Arch Med Sci,2021,18(1):206-222.
[15] WANG Q,GE X,ZHANG J,et al.Effect of lncRNA WT1-AS regulating WT1 on oxidative stress injury and apoptosis of neurons in Alzheimer's disease via inhibition of the miR-375/SIX4 axis[J].Aging(Albany NY),2020,12(23):23974-23995.
[16] ASHMWE M,POSA K,RVHRNÖßL A,et al.Effects of extracorporeal shockwave therapy on functional recovery and circulating miR-375 and miR-382-5p after subacute and chronic spinal cord contusion injury in rats[J].Biomedicines,2022,10(7):1630-1651.
[17] CAO Y,TAN X,LU Q,et al.miR-590-3 and SP1 promote neuronal apoptosis in patients with Alzheimer's disease via AMPK signaling pathway[J].Contrast Media Mol Imaging,2021,2021:1-12.
[18] RAI S N,TIWARI N,SINGH P,et al.Therapeutic potential of vital transcription factors in Alzheimer's and Parkinson's disease with particular emphasis on transcription factor EB mediated autophagy[J].Front Neurosci,2021,15:1-20.
[19] CAI L J,TU L,LI T,et al.Up-regulation of microRNA-375 ameliorates the damage of dopaminergic neurons,reduces oxidative stress and inflammation in Parkinson's disease by inhibiting SP1[J].Aging(Albany NY),2020,12(1):672-689.
[20] XU C,YIN H,JIANG X,et al.Silencing long noncoding RNA LINC01138 inhibits aerobic glycolysis to reduce glioma cell proliferation by regulating the microRNA-375/SP1 axis[J].Mol Med Rep,2021,24(6):846-859.
[21] DE MEDEIROS L M,DE BASTIANI M A,RICO E P,et al.Cholinergic differentiation of human neuroblastoma SH-SY5Y cell line and its potential use as an in vitro model for Alzheimer's disease studies[J].Mol Neurobiol,2019,56(11):7355-7367.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录