NEAT1、miR-27a-3p在阿尔茨海默病患者血清和脑脊液中的表达关系

doi:10.19723/j.issn.1671-167X.2024.02.002

摘要/Abstract

摘要：

目的: 探究长链非编码RNA核富含丰富的转录本1(long chain non-coding RNA nuclear-enriched abundant transcript 1，LncRNA NEAT1)、miR-27a-3p在阿尔茨海默病(Alzheimer disease，AD)患者血清和脑脊液中的表达关系及意义。方法: 选择2019年10月至2021年9月天津市第五中心医院神经内科收治的AD患者66例作为病例组，根据临床痴呆评定量表(clinical dementia rating，CDR)评分分为轻度组(≤1分，n=41)与中重度组(＞1分，n=25)；另取同期门诊其他就诊患者血清和脑脊液标本66例志愿者作为对照组。收集所有受试者的一般资料并评估认知程度，采用实时荧光定量PCR检测血清和脑脊液miR-27a-3p、NEAT1表达水平，酶联免疫吸附试验检测脑脊液β-淀粉样前体蛋白裂解酶1(β-site amyloid precursor protein cleaving enzyme 1，BACE1)、β淀粉样蛋白(amyloid β，Aβ)40和Aβ42水平，采用Spearman法分析血清miR-27a-3p、NEAT1水平与简易精神状态检测量表的相关性，采用Pearson法分析血清miR-27a-3p、NEAT1水平与Aβ沉积平均标准摄取值比率(standardized uptake value ratio，SUVR)及脑脊液miR-27a-3p、NEAT1、BACE1、Aβ42、Aβ40水平的相关性。结果: MMSE评分[21 (17，25)，9(7，11) vs. 27 (21，34)]、MoCA评分[17 (12，21)，10 (7，13) vs. 27 (21，31)]、血清miR-27a-3p水平(0.55±0.13，0.46±0.06 vs. 0.97±0.22)、脑脊液miR-27a-3p(0.48±0.10，0.35±0.10 vs. 1.03±0.31)、Aβ42水平[(303.55±36.77) ng/L，(231.45±34.14) ng/L vs.(499.99±53.63) ng/L]及Aβ42/Aβ40比值(0.030±0.008，0.022±0.007 vs. 0.048±0.010)轻度组、中重度组AD患者均低于对照组(P均＜0.05)，且中重度组AD患者较轻度组低(P均＜0.05)；血清NEAT1水平(2.31±0.64，3.13±0.76 vs. 1.05±0.20)、SUVR(1.50±0.29，1.76±0.52 vs. 0.74±0.15)及脑脊液NEAT1(3.51±1.24，4.30±1.65 vs. 1.01±0.23)、BACE1水平[(55.78±5.98) μg/L，(72.32±16.08) μg/L vs. (21.39±3.73) μg/L]轻度组、中重度组AD患者均高于对照组(P均＜0.05)，且中重度组AD患者较轻度组高(P均＜0.05)。AD患者血清NEAT1水平与SUVR及脑脊液NEAT1、BACE1呈正相关(r=0.350，0.606，0.341，P＜0.05)，与MMSE评分、MoCA评分呈负相关(r=-0.473，-0.482，P均＜0.05)；血清miR-27a-3p水平与脑脊液miR-27a-3p水平、MMSE评分、MoCA评分呈正相关(r=0.695，0.424，0.412，P＜0.05)，与SUVR及脑脊液BACE1水平呈负相关(r=-0.521、-0.447，P均＜0.05)。结论: NEAT1、miR-27a-3p在AD患者血清及脑脊液中表达趋势具有一致性，NEAT1水平均升高，miR-27a-3p水平均降低，二者水平呈负相关，与AD患者脑中Aβ沉积程度有关，并参与AD的病情进展。

关键词: 阿尔茨海默病, 长链非编码RNA核富含丰富的转录本1, miR-27a-3p, β淀粉样蛋白, 相关性

Abstract:

Objective: To explore the expression relationship and significance of long chain non-coding RNA nuclear-enriched abundant transcript 1 (LncRNA NEAT1) and miR-27a-3p in serum and cerebrospinal fluid of patients with Alzheimer disease (AD). Methods: Sixty-six AD patients received by the department of neurology of our hospital from October 2019 to September 2021 were gathered, according to the clinical dementia rating scale score, they were grouped into mild group (≤1 point, n=41) and moderate-to-severe group (>1 point, n=25). Another 66 cases of serum and cerebrospinal fluid samples from outpatient physical examination personnel were regarded as the control group. The general information on all subjects was recorded and cognition was assessed; real-time quantitative PCR was performed to measure the expression levels of miR-27a-3p and NEAT1 in serum and cerebrospinal fluid; enzyme-linked immunosorbent assay was performed to measure the protein levels of β-amyloid precursor protein cleaving enzyme 1 (BACE1), β-amyloid (Aβ) 40 and Aβ42 in cerebrospinal fluid; Spearman' s method was performed to analyze the correlation of serum miR-27a-3p and NEAT1 levels with mini-mental state examination (MMSE) and montreal cognitive assessment (MoCA) scores; Pearson method was performed to analyze the correlation between serum miR-27a-3p and NEAT1 levels and Aβ deposition standard uptake value ratio (SUVR) and cerebrospinal fluid miR-27a-3p, NEAT1, BACE1, Aβ42 and Aβ40 levels. Results: The MMSE score [21 (17, 25), 9(7, 11) vs. 27 (21, 34)], MoCA score [17 (12, 21), 10 (7, 13) vs. 27 (21, 31)], serum miR-27a-3p level (0.55±0.13, 0.46±0.06 vs. 0.97±0.22), cerebrospinal fluid miR-27a-3p (0.48±0.10, 0.35±0.10 vs. 1.03±0.31), Aβ42 levels [(303.55±36.77) ng/L, (231.45±34.14) ng/L vs. (499.99±53.63) ng/L] and Aβ42/Aβ40 ratio (0.030±0.008, 0.022±0.007 vs. 0.048±0.010) of AD patients in mild group and moderate-to-severe group were all lower than those in the control group, and the moderate-to-severe group were lower than the mild group (all P < 0.05); the serum NEAT1 level (2.31±0.64, 3.13±0.76 vs. 1.05±0.20), SUVR (1.50±0.29, 1.76±0.52 vs. 0.74±0.15), and cerebrospinal fluid NEAT1 (3.51±1.24, 4.30±1.65 vs. 1.01±0.23) and BACE1 levels [(55.78±5.98) μg/L, (72.32±16.08) μg/L vs. (21.39±3.73) μg/L] were higher than those in the control group, and the moderate-to-severe group were higher than the mild group (all P < 0.05). Serum NEAT1 level in AD patients was positively correlated with SUVR, cerebrospinal fluid NEAT1 and BACE1 (r=0.350, 0.606, 0.341, P < 0.05), and negatively correlated with MMSE score and MoCA score (r=-0.473, -0.482, all P < 0.05); serum miR-27a-3p level was positively correlated with cerebrospinal fluid miR-27a-3p level, MMSE score and MoCA score (r=0.695, 0.424, 0.412, all P < 0.05), and negatively correlated with SUVR and cerebrospinal fluid BACE1 level (r=-0.521, -0.447, all P < 0.05). Conclusion: The expression trends of NEAT1 and miR-27a-3p in the serum and cerebrospinal fluid of AD patients are consistent, the level of NEAT1 is increased, and the level of miR-27a-3p is decreased. The levels of the two are negatively correlated, which is related to the degree of Aβ deposition in the brain of AD patients and is involved in the progression of AD.

Key words: Alzheimer disease(AD), Long chain non-coding RNA nuclear-enriched abundant transcript 1 (LncRNA NEAT1), miR-27a-3p, Amyloid β (Aβ), Correlation

中图分类号:

R749.16

何丽杰,张春艳,王静. NEAT1、miR-27a-3p在阿尔茨海默病患者血清和脑脊液中的表达关系[J]. 北京大学学报（医学版）, 2024, 56(2): 207-212.

Lijie HE,Chunyan ZHANG,Jing WANG. Expression relationship and significance of NEAT1 and miR-27a-3p in serum and cerebrospinal fluid of patients with Alzheimer disease[J]. Journal of Peking University (Health Sciences), 2024, 56(2): 207-212.

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Group	n	Age/years, $\bar x \pm s$	Gender, male/female	Educational attainment/years, $\bar x \pm s$	Alcohol, n(%)	Smoking, n(%)	Hypertension, n(%)	Diabetes mellitus, n(%)	MMSE scores, M(P₂₅, P₇₅)	MoCA scores, M(P₂₅, P₇₅)
Control	66	69.45±7.36	39/27	7.65±2.13	15 (22.73)	27 (40.91)	25 (37.88)	15 (22.73)	27 (21，34)	27 (21，31)
Mild	41	71.31±7.52	23/18	7.98±2.26	9 (21.95)	19 (46.34)	13 (31.71)	7 (17.07)	21 (17，25)^#	17 (12，21)^#
Moderate-to- severe	25	72.57±6.83	15/10	6.92±2.05	4 (16.00)	13 (52.00)	12 (48.00)	6 (24.00)	9 (7，11)^#*	10 (7，13)^#*
F/χ²/H		1.914	0.128	1.894	0.510	0.967	1.752	0.627	66.379	66.553
P		0.152	0.938	0.154	0.775	0.617	0.416	0.731	< 0.001	< 0.001

Group	n	NEAT1/GAPDH	miR-27a-3p/U6
Control	66	1.05±0.20	0.97±0.22
Mild	41	2.31±0.64^#	0.55±0.13^#
Moderate-to-severe	25	3.13±0.76^#*	0.46±0.06^#*
F		180.359	114.371
P		< 0.001	< 0.001

Group	n	NEAT1/GAPDH	miR-27a-3p/U6	BACE1/(μg/L)	Aβ40/(ng/L)	Aβ42/(ng/L)	SUVR	Aβ42/Aβ40
Control	66	1.01±0.23	1.03±0.31	21.39±3.73	10708.99±1979.17	499.99±53.63	0.74±0.15	0.048±0.010
Mild	41	3.51±1.24^#	0.48±0.10^#	55.78±5.98^#	10477.95±2192.92^#	303.55±36.77^#	1.50±0.29^#	0.030±0.008^#
Moderate- to-severe	25	4.30±1.65^#*	0.35±0.10^#*	72.32±16.08^#*	11019.63±2807.62^#*	231.45±34.14^#*	1.76±0.52^#*	0.022±0.007^#*
F		132.816	113.447	444.023	0.464	415.490	144.609	98.231
P		< 0.001	< 0.001	< 0.001	0.630	< 0.001	< 0.001	< 0.001

Serum NEAT1	SUVR	NEAT1	BACE1	Aβ42	Aβ40	MMSE scores	MoCA scores
r	0.350	0.606	0.341	－0.096	－0.115	－0.473	－0.482
P	< 0.001	< 0.001	0.005	0.445	0.358	< 0.001	< 0.001

Serum miR-27a-3p	SUVR	miR-27a-3p	BACE1	Aβ42	Aβ40	MMSE scores	MoCA scores
r	－0.521	0.695	－0.447	0.074	－0.047	0.424	0.412
P	< 0.001	< 0.001	< 0.001	0.554	0.710	< 0.001	< 0.001