[1]丁 锦a,贯 芳b,梁小涵c,等.急性冠脉综合征患者血浆外泌体NEAT1，miR-204 和MMP-9 的表达水平及临床意义[J].现代检验医学杂志,2023,38(01):59-65.[doi:10.3969/j.issn.1671-7414.2023.01.012]
　DING Jina,GUAN Fangb,LIANG Xiao-hanc,et al.Expression Levels and Clinical Significance of Plasma Exosomal NEAT1, miR-204 and MMP-9 in Patients with Acute Coronary Syndrome[J].Journal of Modern Laboratory Medicine,2023,38(01):59-65.[doi:10.3969/j.issn.1671-7414.2023.01.012]
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急性冠脉综合征患者血浆外泌体NEAT1，miR-204 和MMP-9 的表达水平及临床意义 ()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第38卷
期数:
2023年01期
页码:
59-65
栏目:
论著
出版日期:
2023-01-15

文章信息/Info

Title:
Expression Levels and Clinical Significance of Plasma Exosomal NEAT1, miR-204 and MMP-9 in Patients with Acute Coronary Syndrome
文章编号:
1671-7414（2023）01-059-07
作者:
丁 锦a; 贯 芳b; 梁小涵c; 陈 旭a
（首都医科大学附属北京世纪坛医院 a. 急诊科；b. 消化内科；c. 胃肠肝胆肿瘤外科，北京100038）
Author(s):
DING Jina; GUAN Fangb; LIANG Xiao-hanc;  CHEN Xua
(a. Department of Emergency; b. Department of Gastroenterology; c. Department of Gastrointestinal and Hepatobiliary Tumor Surgery, Beijing Shijitan Hospital Affiliated to Capital Medical University, Beijing 100038, China)
关键词:
血浆外泌体; 富含核富集的转录物1（NEAT1）; 微小核糖核酸-204（miR-204); 基质金属蛋白酶-9(MMP -9); 急性冠脉综合征
分类号:
R541.4；R392.11
DOI:
10.3969/j.issn.1671-7414.2023.01.012
文献标志码:
A
摘要:
目的 检验急性冠脉综合征（acute coronary syndrome，ACS）患者血浆外泌体富含核富集的转录物1 (Nucleraenrichedautosomal transcript，NEAT1)，微小核糖核酸（microRNA，miR)-204 和基质金属蛋白酶 (metalloproteinase，MMP) -9 的表达水平，并研究其对ACS的诊断价值，分析其与冠脉病变严重程度的相关性。方法 选取2020 年5 月～ 2021年5 月于首都医科大学附属北京世纪坛医院诊断为ACS 的120 例患者作为实验组，其中急性心肌梗死（acute myocardialinfarction，AMI）患者72 例，不稳定心绞痛（unstable angina，UA）患者48 例；另选取行冠脉造影检查无明显狭窄的108 例体检者作为Control 组。提取所有研究对象的血浆外泌体，并采用透射电镜和Western blot 鉴定该外泌体。应用试剂盒提取血浆外泌体总RNA，采用qRT-PCR 法检测外泌体NEAT1 和miR-204 的表达水平，采用Western blot 检测外泌体MMP-9 蛋白表达水平。应用Pearson 相关分析明确外泌体NEAT1，miR-204 和MMP-9 水平与病变严重程度评分（Gensini 评分）之间的相关性。Logistic 回归模型判断外泌体NEAT1，miR-204 和MMP-9 是否可作为诊断ACS 的独立危险因素。绘制受试者工作特征（ROC）曲线，根据曲线下面积（AUC）分析三者表达水平对ACS 病变严重程度的预测价值。结果 透射电镜观察到，血浆外泌体呈椭圆形双层膜囊泡结构，Western blot 结果显示，CD63 和CD81 蛋白在外泌体中呈高表达。Control 组 NEAT1，miR-204 和MMP-9 表达水平分别为0.62±0. 08，1.02±0. 20 和0.97±0.15，UA 组患者NEAT1，miR-204 表达水平分别为0.65±0.11，1.05±0.18，与Control 组比较差异无统计学意义（t=2.790，3.225，P ＞ 0.05），而MMP-9 表达水平（1.15±0. 20）较Control 组明显升高，差异有统计学意义（t=13.682，P ＜ 0.05）；AMI 组患者血浆外泌体NEAT1，miR-204 和MMP-9 表达水平分别为0.98±0.15，1.22±0.23 和1.37±0.25，较Control组明显升高，差异均有统计学意义（t=12.112, 9.885, 21.530, 均P ＜ 0.05）。Pearson 相关性分析显示，AMI 患者外泌体NEAT1，MMP-9与Gensini 评分呈中度正相关， miR-204 与Gensini 评分呈弱正相关，差异有统计学意义（r=0.179 ～ 0.548，均P ＜ 0.05）。UA 患者外泌体NEAT1 与Gensini 评分呈弱正相关（r=0.207，P=0.032），MMP-9 与Gensini 评分呈中度正相关（r=0.574，P ＜ 0.05），但miR-204 与Gensini 评分无相关性（r=0.108，P=0.465）。Logistic 回归分析结果显示，外泌体NEAT1，miR-204 和MMP-9 均可作为AMI 预测的独立危险因素，但外泌体miR-204 不可作为预测UA 的独立危险因素。ROC 结果显示，AMI 组外泌体NEAT1，miR-204 和MMP-9 水平及联合检测对应AUC 分别为0.821，0.702，0.750 和0.905，UA 组外泌体NEAT1，miR-204 和MMP-9 水平及联合检测对应AUC 分别为0.776，0.682，0.718 和0.883，三者对AMI 的诊断价值均高于UA，且联合检测具有更高诊断价值。结论 血浆外泌体NEAT1，miR-204 和MMP-9 具有预测冠状动脉病变严重程度的潜力，并且三者联合检测对AMI具有较高诊断价值，可作为辅助诊断AMI的潜在标志物。
Abstract:
Objective To investigate the expression level of plasma exosome nuclera-enriched autosomal transcript(NEAT1)， microRNA-204（miR-204) and metalloproteinase(MMP)-9 in patients with acute coronary syndrome (ACS), and study its diagnostic value for ACS and its correlation with the severity of coronary lesions. Methods 120 patients with ACS diagnosed in the Department of Cardiology of Beijing Shijitan Hospital Affiliated to Capital Medical University from May 2020 to May 2021 were selected as the experimental group, including 72 patients with acute myocardial infarction (AMI) and 48 patients with unstable angina pectoris (UA). In addition, 108 patients hospitalized in the department in the same period and found no obvious coronary stenosis by coronary angiography were selected as the control group. The plasma exosomes of all subjects were extracted and identified by transmission electron microscopy and Western blot. The total RNA of the above exosomes was extracted by the kit, the expression levels of exosomal NEAT1 and miR-204 were detected by qRT-PCR, and the protein expression levels of exosomal MMP-9 were detected by Western blot. Pearson correlation analysis was used to clarify the correlation between exosomal NEAT1, miR-204, MMP-9 levels and lesion severity score (Gensini score). Logistic regression model to determine whether exosomal NEAT1, miR-204 and MMP-9 could be used as an independent risk factor for the diagnosis of ACS. Draw the receiver operating characteristic (ROC) curve, and analyze the predictive value of the three expression levels on the severity of ACS lesions according to the area under the curve AUC. Results Transmission electron microscope observed that the plasma exosomes had an oval double-membrane vesicle structure. Western blot results showed that CD63 and CD81 proteins were significantly expressed in exosomes. The expressions of NEAT1, miR-204 and MMP-9 in the Control group were 0.62±0.08，1.02±0.20 and 0.97±0.15, respectively. The expressions of NEAT1 and miR-204 in the UA group were 0.65±0.11 and 1.05±0.18, respectively, with no significant difference compared with the Control group (t=2.790, 3.225, P ＞ 0.05), while the expression level of MMP-9 (1.15±0.20) was significantly higher than that in the Control group, the difference was statistically significant（t=13.682, P ＜ 0.05）. The expressions level of plasma exosomal NEAT1, miR-204 and MMP-9 in the AMI group were 0.98±0.15，1.22±0.23 and 1.37±0.25, respectively, which were significantly higher than those in the Control group, and the differences were statistically significant (t=9.885 ～ 21.530, all P ＜ 0.05). Pearson correlation analysis showed that exosomal NEAT1, MMP-9 and Gensini scores were moderately positively correlated in AMI patients, and miR-204 was weakly positively correlated with Gensini scores, and the differences were statistically significant (r=0.179 ～0.548, all P ＜ 0.05), exosomal NEAT1 in UA patients was weakly positively correlated with Gensini score (r=0.207, P=0.032), and MMP-9 was moderately positively correlated with Gensini score (r=0.574, P ＜ 0.05), but there was no significant difference between miR-204 and Gensini score (r=0.108, P=0.465). Logistic regression analysis showed that exosomal NEAT1, miR-204 and MMP-9 could be used as independent risk factors for AMI, however, exosomal miR-204 could not be used as an independent risk factor for UA prediction. The ROC results showed that the levels of exosomal NEAT1, miR-204 and MMP-9 in the AMI group and the corresponding AUC of the combined detection were 0.821, 0.702, 0.750 and 0.905, respectively. The levels of exosomal NEAT1, miR-204 and MMP-9 in the UA group and the corresponding AUCs of the combined detection were 0.776, 0.682, 0.718 and 0.883, respectively. The diagnostic value of the three for AMI was higher than that of UA, and the combined detection had a higher diagnostic value. Conclusion Plasma exosomal NEAT1, miR-204 and MMP-9 had the potential to predict the severity of coronary artery lesions, and the combined detection of the three has high diagnostic value for AMI, and could be used as potential markers for auxiliary diagnosis of AMI.

参考文献/References:

[1] 刘永玲, 罗厚龙, 刘行超, 等．血清SOD 和hsCRP等指标在急性冠脉综合征中的应用及相关性分析[J]. 现代检验医学杂志, 2017, 32(6)：115-117, 121．
LIU Yongling, LUO Houlong, LIU Xingchao, et al.Level and correlationship of serum SOD and hsCRP and other indicators in acute coronary syndrome [J].Journal of Modern Laboratory Medicine, 2017, 32(6):115-117, 121.
[2] LEE J, KIISKINEN T, MARS N, et al. Clinical conditions and their impact on utility of genetic scores for prediction of acute coronary syndrome[J].Circulation. Genomic and Precision Medicine, 2021,14(4): e003283.
[3] ZAMANI P, FEREYDOUNI N, BUTLER A E, et al.The therapeutic and diagnostic role of exosomes in cardiovascular diseases[J]. Trends in Cardiovascular Medicine, 2019, 29(6): 313-323.
[4] ZHANG Yunyuan, LUN Limin, LI Hui, et al. The value of lncRNA NEAT1 as a prognostic factor for survival of cancer outcome: a meta-analysis[J]. Scientific Reports,2017, 7(1): 13080.
[5] KOYAMA R, MANNIC T, ITO J, et al. MicroRNA-204 is necessary for aldosterone-stimulated T-type calcium channel expression in cardiomyocytes[J]. International Journal of Molecular Sciences, 2018, 19(10): 2941.
[6] SANTANA I V, TANUS-SANTOS J E. Serum or plasma matrix metalloproteinase (MMP)-9 levels and cardiovascular diseases[J]. Journal of Cardiovascular Translational Research, 2018, 11(6): 524-525.
[7] CREA F, LIBBY P. Acute coronary syndromes: the way forward from mechanisms to precision treatment[J].Circulation, 2017, 136(12): 1155-1166.
[8] HENNING R J. Cardiovascular exosomes and microRNAs in cardiovascular physiology and pathophysiology[J]. Journal of Cardiovascular Translational Research, 2021, 14(2): 195-212.
[9] MA Min, HUI Jie, ZHANG Qiyin, et al. Long noncoding RNA nuclear-enriched abundant transcript 1 inhibition blunts myocardial ischemia reperfusion injury via autophagic flux arrest and apoptosis in streptozotocin-induced diabetic rats[J]. Atherosclerosis,2018, 277(9): 113-122.
[10] WANG Lei, XIA Jingwen, KE Zunping, et al. Blockade of NEAT1 represses inflammation response and lipid uptake via modulating miR-342-3p in human macrophages THP-1 cells [J]. Journal of Cellular Physiology, 2019, 234(4): 5319-5326.
[11] LUO Man, SUN Qingsong, ZHAO Hongmei, et al.Long noncoding RNA NEAT1 sponges miR-495-3p to enhance myocardial ischemia-reperfusion injury via MAPK6 activation[J]. Journal of Cellular Physiology,2020, 235(1): 105-113.
[12] LU Yaoyong, LI Tao, WEI Ganbao, et al. The long non-coding RNA NEAT1 regulates epithelial to mesenchymal transition and radioresistance in through miR-204/ZEB1 axis in nasopharyngeal carcinoma[J].Tumor Biology, 2016, 37(9): 11733-11741.
[13] LIU Jing, LIU Yong, WANG Feng, et al. MiR-204:molecular regulation and role in cardiovascular and renal diseases[J]. Hypertension, 2021, 78(2): 270-281.
[14] DU Youyou, LIU Ganghui, ZHAO Luosha, et al.Protective effect of miR-204 on doxorubicin-induced cardiomyocyte injury via HMGB1[J]. Oxidative Medicine and Cellular Longevity, 2020, 2020: 8819771.
[15] 吴静, 燕成英, 温元善, 等．血清miR-9,miR-204 水平对PCI 术后ACS 患者预后的评估价值[J]．心血管康复医学杂志, 2021, 30(2)：143-147．
WU Jing, YAN Chengying, WEN Yuanshan, et al. The value of the expressions of miR-9 and miR-204 in the prognosis of acute coronary syndrome [J]. Chinese Journal of Cardiovascular Rehabilitation Medicine,2021, 30(2):143-147.
[16] LUAN Wenkang, QIAN Yao, NI Xin, et al. MiR-204-5p acts as a tumor suppressor by targeting matrix metalloproteinases-9 and B-cell lymphoma-2 in malignant melanoma[J]. Onco Targets & Therapy,2017, 10(2): 1237-1246.
[17] WANG Xi, KHALIL R A. Matrix metalloproteinases,vascular remodeling, and vascular disease[J]. Advances in Pharmacology (San Diego, Calif.), 2018, 81: 241-330.
[18] NANDI S S, KATSURADA K, SHARMA N M, et al.MMP-9 inhibition increases autophagic flux in chronic heart failure[J]. American Journal of Physiology-Heart and Circulatory Physiology, 2020, 319(6):H1414-H1437.
[19] 陈俊, 廖应英, 孙泽群．基质金属蛋白酶-9 与心房颤动的关系及对持续性心房颤动患者复律治疗后复发的预测价值[J]. 中国循环杂志, 2017, 32(1)：67-71.
CHEN Jun, LIAO Yingying, SUN Zequn, et al.Relationship between matrix metalloproteinase-9 level and atrial fibrillation with its predictive value of AF recurrence in persistent AF patients after cardio-version[J]. Chinese Circulation Journal, 2017, 32(1): 67-71.
[20] YU Yang , WANG Leilei, LIU Tong , et al . MicroRNA-204 suppresses trophoblast-like cell invasion by targeting matrix metalloproteinase-9[J]. Biochemical and Biophysical Research Communications, 2015, 463(3): 285-291.

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备注/Memo

备注/Memo:
作者简介：丁锦(1995-)，女，本科，主治医师，研究方向：急性心肌梗死，E-mail：dingjin@bjsjth.cn。
通讯作者：陈旭（1980-），女，本科，副主任医师，研究方向：急性心肌梗死。
更新日期/Last Update: 2023-01-15