冠心病患者心外膜脂肪组织和血浆中脂联素相关miR-371b-5p表达及其对脂肪细胞因子分泌的影响

doi:10.13481/j.1671-587x.20190329

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (03): 643-650.doi: 10.13481/j.1671-587x.20190329

• 临床研究 • 上一篇

冠心病患者心外膜脂肪组织和血浆中脂联素相关miR-371b-5p表达及其对脂肪细胞因子分泌的影响

杜雅彦¹, 刘洋², 卢沐³, 怀施涛¹, 吕作利¹, 魏育涛⁴

1. 石河子大学医学院新疆民族与地方病教育部重点实验室, 新疆石河子 832008;
2. 首都医科大学附属朝阳医院京西院区急诊科, 北京 100020;
3. 石河子大学医学院附属第一医院心胸外二科, 新疆石河子 832008;
4. 广东省第二人民医院心胸外科, 广东广州 510000

收稿日期:2018-08-08 发布日期:2019-06-05
通讯作者: 魏育涛,教授,硕士研究生导师(Tel:0993-2858456,E-mail:wytfwb@126.com)) E-mail:wytfwb@126.com
作者简介:杜雅彦(1989),女,甘肃省张掖市人,在读医学硕士,主要从事冠心病基础和临床方面的研究。
基金资助:
国家自然科学基金资助课题（81460059）

Expressions of adiponectin-related miR-371b-5p in epicardial adipose tissue and plasma of patients with coronary heart disease and its effect on secretion of adipocytokines

DU Yayan¹, LIU Yang², LU Mu³, HUAI Shitao¹, LYU Zuoli¹, WEI Yutao⁴

1. Key Laboratory of Xinjiang Ethnic and Endemic Diseases, Ministry of Education, School of Medical Sciences, Shihezi University, Shihezi 832008, China;
2. Department of Cardiothoracic Surgery, Guangdong Second Provincial General Hospital, Guangzhou 510000, China;
3. Affiliated Beijing Chaoyang Hospital, Capital University of Medical Sciences, Beijing 100020, China;
4. Second Department of Thoracic and Cardiovascular Surgery, First Affiliated Hospital, School of Medical Sciences, Shihezi University, Shihezi 832008, China

Received:2018-08-08 Published:2019-06-05

摘要/Abstract

摘要： 目的：探讨冠心病（CAD）患者心外膜脂肪组织（EAT）和血浆中脂联素（APN，又称ADIPOQ）相关差异表达miR-371b-5p对脂肪细胞因子APN、Kruppel样因子4（KLF4）、白细胞介素6（IL-6）和单核细胞因子趋化蛋白1（MCP-1）分泌的影响，阐明miR-371b-5p在CAD中的作用机制。方法：收集CAD （CAD组，n=34）和非CAD （non-CAD组，n=16）患者的EAT和血液标本，采用实时荧光定量PCR （qRT-PCR）法检测2组患者EAT和血浆中miR-371b-5p的表达水平。将诱导成功的3T3-L1前体脂肪细胞分为空白对照组（不做任何处理）、miR-371b-5p过表达组（转染miR-371b-5p模拟物）和miR-371b-5p抑制剂组（转染miR-371b-5p抑制剂）。qRT-PCR法检测转染24 h后各组3T3-L1前脂肪细胞中APN、KLF4、IL-6和MCP-1 mRNA表达水平，并对miR-371b-5p进行生物信息学分析。结果：与non-CAD组比较，CAD组患者EAT和血浆中miR-371b-5p表达水平均升高（P<0.01）。与空白对照组比较，miR-371b-5p过表达组3T3-L1前脂肪细胞中APN和KLF4 mRNA表达水平降低（P=0.043，P=0.045），IL-6和MCP-1mRNA表达水平升高（P=0.037，P=0.041）；miR-371b-5p抑制剂组3T3-L1前脂肪细胞中APN和KLF4mRNA表达水平升高（P=0.025，P=0.027），IL-6和MCP-1mRNA表达水平降低（P=0.039，P=0.041）。miR-371b-5p预测靶基因富集于多个生物学功能及过程，KEGG通路主要富集于脂肪细胞因子等信号通路；蛋白互作，miR-371b-5p预测靶基因APN、瘦素（LEP）和AKT1编码蛋白位于互作网络中核心位置。结论：CAD患者EAT中过表达miR-371b-5p转染成熟的3T3-L1脂肪细胞后可使细胞的抗炎因子表达水平降低，炎性因子表达水平升高，miR-371b-5p有望成为CAD治疗的新靶点。

关键词: 冠心病, 心外膜脂肪组织, 微小RNA, 脂肪细胞因子, 生物信息学分析

Abstract: Objective:To investigate the effects of adiponectin (APN,also known as ADIPOQ)-related miR-371b-5p with differential expressions in the epicardial adipose tissue (EAT) and plasma in the patients with coronary artery disease (CAD) on the secretions of APN, Kruppel-like factor 4 (KLF4), interleukin-6 (IL-6) and monocyte chemokine 1 (MCP-1), and to elucidate the mechanism of action of miR-371b-5p in CAD. Methods:The EAT and blood samples were collected from the patients with CAD (CAD group,n=34) and non-CAD (non-CAD group,n=16). Real-time quantitative PCR (qRT-PCR) was used to detect the expression levels of miR-371b-5p in the EAT and plasma of the patients in two groups. The successfully induced 3T3-L1 preadipocytes were divided into(without any treatment), miR-371b-5p over-expression group (transfected with miR-371b-5p mimics) and miR-371b-5p inhibitor group (transfected with miR-371b-5p inhibitor). The relative expression levels of APN, KLF4, IL-6 and MCP-1 mRNA in the 3T3-L1 preadipocytes were detected by qRT-PCR method. The miR-371b-5p was analyzed by bioinformatics. Results:The expression levels of miR-371b-5p in the EAT and plasma of the patients in CAD group were higher than those in non-CAD group(P<0.01). Compared with blank control group,the expression levels of APN and KLF4 mRNA in 3T3-L1 preadipocytes in miR-371b-5p over-expression group were decreased(P=0.043, P=0.045), and the expression levels of IL-6 and MCP-1 mRNA in 3T3-L1 preadipocytes in miR-371b-5p over-expression group were increased (P=0.037, P=0.041). Compared with blank control group, the expression levels of APN and KLF4 in 3T3-L1 preadipocytes in miR-371b-5p inhibitor group were increased(P=0.025, P=0.027), and the expression levels of IL-6 and MCP-1 mRNA in 3T3-L1 preadipocytes in miR-371b-5p inhibitor group were decreased (P=0.039, P=0.041). The target genes predicted by miR-371b-5p were enriched in multiple biological functions and processes. The KEGG pathway was mainly enriched in the signaling pathways such as adipocytokines. The protein interaction results showed that the target genes APN,leptin(LEP) and AKT1 encoded proteins predicted by miR-371b-5p were located at the core of the interaction network. Conclusion:The expression levels of anti-inflammatory factors in the cells with over-expression of miR-371b-5p in EAT of the patients with CAD are decreased, the expression levels of inflammatory factors are increased,and miR-371b-5p is expected to become a new target for CAD treatment.

Key words: coronary artery disease, epicardial adipose tissue, miRNA, adiponectin, bioinformatics analysis

中图分类号:

R541.4

杜雅彦, 刘洋, 卢沐, 怀施涛, 吕作利, 魏育涛. 冠心病患者心外膜脂肪组织和血浆中脂联素相关miR-371b-5p表达及其对脂肪细胞因子分泌的影响[J]. 吉林大学学报(医学版), 2019, 45(03): 643-650.

DU Yayan, LIU Yang, LU Mu, HUAI Shitao, LYU Zuoli, WEI Yutao. Expressions of adiponectin-related miR-371b-5p in epicardial adipose tissue and plasma of patients with coronary heart disease and its effect on secretion of adipocytokines[J]. Journal of Jilin University(Medicine Edition), 2019, 45(03): 643-650.

参考文献

[1] CARVALHO E, LOPASCHUK G D, BØRSHEIM E, et al. Reply to Katlandur, Ozbek, and Keser[J]. Am J Physiol Endocrinol Metab, 2016, 310(10):E863.
[2] VINKHUYZEN AA, WRAY NR, YANG J, et al. Estimation and partition of heritability in human populations using whole-genome analysis methods[J]. Annu Rev Genet, 2013, 47:75-95.
[3] PAUL P, CHAKRABORTY A, SARKAR D, et al. Interplay between miRNAs and human diseases[J]. J Cell Physiol, 2018, 233(3):2007-2018.
[4] BORK S, HORN P, CASTOLDI M, et al. Adipogenic differentiation of human mesenchymal stromal cells is down-regulated by microRNA-369-5p and up-regulated by microRNA-371[J]. J Cell Physiol, 2011, 226(9):2226-2234.
[5] LIU Y, FU W, LU M, et al. Role of miRNAs in epicardial adipose tissue in CAD patients with T2DM[J]. BioMed Res Int, 2016, 2016(16):e29236.
[6] IACOBELLIS G. Epicardial and pericardial fat:close, but very different[J]. Obesity (Silver Spring, Md), 2009, 17(4):625; author reply 6-7.
[7] WU Y, ZHANG A, HAMILTON D J, et al. Epicardial fat in the maintenance of cardiovascular health[J]. Methodist DeBakey Cardiovasc J, 2017, 13(1):20-24.
[8] 魏育涛, 付文博, 刘诚, 等. 心外膜脂肪厚度与冠心病的相关性研究[J]. 临床心血管病杂志, 2014, 30(9):758-761.
[9] KUMADA M, KIHARA S, OUCHI N, et al. Adiponectin specifically increased tissue inhibitor of metalloproteinase-1 through interleukin-10 expression in human macrophages[J]. Circulation, 2004, 109(17):2046-2049.
[10] LOVREN F, PAN Y, QUAN A, et al. Adiponectin primes human monocytes into alternative anti-inflammatory M2 macrophages[J]. Am J Physiol Heart Circ Physiol, 2010, 299(3):H656-663.
[11] MAHADEV K, WU X, DONNELLY S, et al. Adiponectin inhibits vascular endothelial growth factor-induced migration of human coronary artery endothelial cells[J]. Cardiovasc Res, 2008, 78(2):376-384.
[12] WANG X, PU H, MA C, et al. Adiponectin abates atherosclerosis by reducing oxidative stress[J]. Med Sci Monit, 2014, 20(1):792-800.
[13] VACCA M, DI EUSANIO M, CARIELLO M, et al. Integrative miRNA and whole-genome analyses of epicardial adipose tissue in patients with coronary atherosclerosis[J]. Cardiovasc Res, 2016, 109(2):228-239.
[14] WANG X, LI J, TANG W, et al. Adiponectin receptor 1-mediated micro RNA-323-3p regulates functions of the MIN6 cell line via the AMP-activated protein kinase/sirtuin-1 pathway[J]. J Int Med Res, 2018, 46(5):1693-1708.
[15] LEI L, ZHOU C, YANG X, et al. Down-regulation of microRNA-375 regulates adipokines and inhibits inflammatory cytokines by targeting AdipoR2 in non-alcoholic fatty liver disease[J]. Clin Exp Pharmacol, 2018, 22(5):e12940.
[16] 刘雅青. IRS-1通过miR-143调节前脂肪细胞分化的作用与机制研究[D].长沙:中南大学, 2014.
[17] POLLAK N M, HOFFMAN M, GOLDBERG I J, et al. Kruppel-like factors:Crippling and un-crippling metabolic pathways[J]. JACC Basic Transl Sci, 2018, 3(1):132-156.
[18] FERRANDI P J, FICO B G, WHITEHURST M, et al. Acute high-intensity interval exercise induces comparable levels of circulating cell-free DNA and interleukin-6 in obese and normal-weight individuals[J]. Life Sci, 2018, 20(2):161-166.
[19] LI X. The association between MCP-1, VEGF polymorphisms and their serum levels in patients with diabetic foot ulcer[J]. Medicine (Baltimore), 2018, 97(24):e10959.
[20] 魏育涛. 心外膜脂肪内分泌紊乱诱导冠状动脉粥样硬化机制的研究[D].武汉:华中科技大学, 2011.
[21] FARR O M, GAVRIELI A, MANTZOROS C S. Leptin applications in 2015:what have we learned about leptin and obesity?[J]. Curr Opin Endocrinol Diabetes Obes, 2015, 22(5):353-359.
[22] KUMARI R, KUMAR S, AHMAD M K, et al. Promoter variants of TNF-alpha rs1800629 and IL-10 rs1800871 are independently associated with the susceptibility of coronary artery disease in north Indian[J]. Cytokine, 2018, 110(1):31-36.
[23] ERSAHIN T, TUNCBAG N, CETIN-ATALAY R. The PI3K/AKT/mTOR interactive pathway[J]. Mol Bio-Systems, 2015, 11(7):1946-1954.
[24] HAN X, ZHONG Z, KOU J, et al. ROS generated by upconversion nanoparticle-mediated photodynamic therapy induces autophagy via PI3K/AKT/mTOR signaling pathway in M1 peritoneal macrophage[J]. Cell Physiol Biochem, 2018, 48(4):1616-1627.

冠心病患者心外膜脂肪组织和血浆中脂联素相关miR-371b-5p表达及其对脂肪细胞因子分泌的影响

Expressions of adiponectin-related miR-371b-5p in epicardial adipose tissue and plasma of patients with coronary heart disease and its effect on secretion of adipocytokines

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