CMTM5基因与冠状动脉粥样硬化性心脏病的关联研究及机制探讨

doi:10.19723/j.issn.1671-167X.2020.06.015

摘要/Abstract

摘要：

目的:探讨趋化素样因子超家族成员5(CKLF-like MARVEL transmembrane domain containing member 5,CMTM5)基因与冠状动脉粥样硬化性心脏病(简称冠心病)发生风险的相关性,及CMTM5基因表达变化对THP-1细胞黏附及迁移能力的影响。方法:采用病例对照研究法,入选700例首都医科大学附属北京世纪坛医院心血管内科的住院患者,采用冠状动脉造影法,将结果提示至少存在一支血管内径狭窄≥50%的患者诊断为冠心病。采用逆转录-聚合酶链反应法(reverse transcription-polymerase chain reaction,RT-PCR)测定CMTM5基因表达,酶联免疫吸附测定法(enzyme linked immunosorbent assay,ELISA)检测入选患者血浆CMTM5水平,Logistic回归方法分析CMTM5基因与冠心病发生风险的相关性。培养人血管内皮细胞(endothelial cells,ECs)及THP-1细胞,采用黏附实验及Transwells迁移实验评价CMTM5基因对THP-1趋化能力的影响。结果:冠心病组患者CMTM5基因mRNA表达量是对照组表达量的3.45倍,明显高于对照组(P<0.05)。冠心病组血浆CMTM5蛋白平均水平为(206.1±26.9) μg/L,明显高于对照组的(125.3±15.2) μg/L(P<0.05)。Logistic回归分析纳入年龄、性别、体重指数、吸烟、高血压、糖尿病、高脂血症等冠心病的易患因素和CMTM5基因,结果提示,CMTM5基因仍与冠心病的发生风险存在显著相关性(P<0.05)。黏附实验及Transwells实验结果均提示,过表达CMTM5 ECs组(EO组)中,THP-1细胞的黏附数量及迁移数量明显高于过表达CMTM5对照组(EO-MOCK组)、正常ECs组(EN组)、低表达CMTM5对照组(ES-MOCK组)和低表达CMTM5 ECs组(ES组), 相反,ES组中THP-1细胞黏附数量及迁移数量明显低于其他4组,差异均有统计学意义(P均<0.01)。结论:CMTM5基因与冠心病的发生发展密切相关,CMTM5基因过表达促进THP-1黏附及迁移能力,从而促进动脉粥样硬化和冠心病的发生发展。

关键词: 动脉粥样硬化, 冠状动脉粥样硬化性心脏病, 趋化素样因子超家族成员5, 基因

Abstract:

Objective: To elucidate the correlation between CKLF-like MARVEL transmembrane domain containing member 5 (CMTM5) gene and the risk of coronary artery disease (CAD), and to detect the effects of CMTM5 gene expression changes on the ability of adhesion and migration of THP-1 cells. Methods: Using case-control method, a total of 700 hospitalized patients in Shijitan Hospital were enrolled in this study. CAD were diagnosed by coronary angiography, which was defined as at least one blood vessel diameter stenosis ≥50% according to the result of coronary angiography. Reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect CMTM5 gene expression; enzyme linked immunosorbent assay (ELISA) method to detect the plasma level of CMTM5; and Logistic regression to analyze CMTM5 genes and the risk of CAD. Human vascular endothelial cells (ECs) and THP-1 cells were cultivated, adhesion and Transwells experiments were used to evaluate the chemotactic capabi-lity of CMTM5 gene on THP-1 cells. Results: In this study, 350 CAD patients matched with 350 control patients were included. RT-PCR results revealed CMTM5 mRNA expression in CAD group was 3.45 times compared with control group, which was significantly higher than that in control group (P<0.05). The levels of CMTM5 plasma protein in CAD group was (206.1±26.9) μg/L, which was significantly higher than that in control group (125.3±15.2) μg/L (P<0.05). After adjusted for the risk factors of age, gender, BMI, smoking, hypertension, diabetes and hyperlipidemia, Logistic regression analysis results indicated that CMTM5 was the susceptibility factors of CAD, which still had significant correlation with CAD (P<0.05). Adhesion and Transwells experiments results revealed that the numbers of adhesion and migration of THP-1 cells in CMTM5 overexpression ECs group (EO group) were significantly higher than that in lenti-mock infected ECs group (EO-MOCK group), non-infected ECs group (EN group), lenti-mock infected ECs group (ES-MOCK group), and CMTM5 suppression ECs group (ES group). On the contrary, the numbers of adhesion and migration of THP-1 cells in ES group were significantly lower than that in the other four groups (P<0.01). Conclusion: CMTM5 gene was closely related to the development of CAD. CMTM5 overexpression promoted the adhesion and migration of THP-1, which might play a part in the mechanisms of atherosclerosis and CAD.

Key words: Atherosclerosis, Coronary artery disease, CMTM5, Gene

中图分类号:

R543.3

刘滕飞,林涛,任利辉,李广平,彭建军. CMTM5基因与冠状动脉粥样硬化性心脏病的关联研究及机制探讨[J]. 北京大学学报（医学版）, 2020, 52(6): 1082-1087.

Teng-fei LIU,Tao LIN,Li-hui REN,Guang-ping LI,Jian-jun PENG. Association between CMTM5 gene and coronary artery disease and the relative mechanism[J]. Journal of Peking University (Health Sciences), 2020, 52(6): 1082-1087.

图/表 5

表1

CAD组和对照组的临床基线资料"

Variables	Control group (n=350)	CAD group (n=350)	P
Age/years, $x -$ ±s	58.2±7.3	58.7±8.2	0.535
Gender/(male/female), n	200/150	200/150	Match
BMI/(kg/m²), $x -$ ±s	25.0±4.0	25.3±5.3	0.525
DM, n (%)	32 (9.1)	79 (22.6)	<0.001^*
Hypertension, n (%)	185 (52.9)	220 (62.9)	0.013^*
Current smoking, n (%)	115 (32.9)	170 (48.6)	<0.001^*
TG/(mmol/L), $x -$ ±s	2.15±1.02	2.24±1.30	0.002^*
TC/(mmol/L), $x -$ ±s	4.72±1.05	5.03±1.61	<0.001^*
LDL-C/(mmol/L), $x -$ ±s	2.52±0.62	2.63±0.83	0.013^*
HDL-C/(mmol/L), $x -$ ±s	1.41±0.44	1.44±0.53	0.286

表1

图1

表2

图2

图3

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Risk factors	B	SE	OR	95%CI		P
Risk factors	B	SE	OR	Lower	Upper	P
Age	-0.960	0.116	0.383	0.305	0.481	<0.001
Gender	0.567	0.067	1.763	1.546	2.010	<0.001
BMI	-0.007	0.008	0.993	0.976	0.971	0.377
Smoking	-0.822	0.070	0.439	0.383	0.504	<0.001
Hypertension	0.350	0.066	1.420	1.247	1.617	<0.001
Diabetes mellitus	0.579	0.052	1.784	1.611	1.975	<0.001
Hyperlipidemia	0.259	0.070	1.296	1.130	1.487	<0.001
CMTM5	-0.044	0.007	0.957	0.943	0.971	0.002