[Frontiers in Bioscience, Landmark, 25, 577-592, Jan 1, 2020]

The miR-145 rs353291 C allele increases susceptibility to atherosclerosis

Yu Lv1, Yihu Yi2, Sujie Jia1, Xiangdong Peng1, Heng Yang4, Ren Guo1,3

1Department of Pharmacy, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China, 2Xiangya School of Medical Science of Central South University, Changsha, Hunan 410013, P.R. China, 3Central of Clinical Pharmacology, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China, 4Department of Neurology, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China

TABLE OF CONTENTS

1. ABSTRACT
2. Introduction
3. Materials and methods
    3.1. Subjects
    3.2. Genotyping
    3.3. Determination of sCD40L, IL-6 and MCP-1 concentrations in the plasma
    3.4. Isolation and culture of human PBMCs
    3.5. Construction of reporter plasmids
    3.6. Real-time PCR analysis
    3.7. Western blotting
    3.8. Monocyte adhesion assay
    3.9. Statistical analysis
4. Results
    4.1. Clinical and laboratory characteristics in the patients and controls
    4.2. Distribution of the frequencies of the rs353291 polymorphism in AS patients and controls
    4.3. Association of the rs353291 polymorphism with the plasma levels of sCD40L, IL-6 and MCP-1
    4.4. Effects of the miR-145 rs353291 polymorphism on miR-145 expression in PBMCs from AS patients and controls
    4.5. Dual Luciferase Reporter Assay of the miR-145 rs353291 polymorphism
    4.6. Effects of the miR-145 rs353291 polymorphism on miR-145, CD40 and MCP-1 expression in PBMCs induced by ox-LDL
    4.7. Effects of the miR-145 rs353291 polymorphism on the NF-κB pathway in THP-1 cells
5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

Atherosclerosis (AS) is one of the leading causes of death, with variable frequencies in different populations. To this end, we examined whether AS is associated with the miR-145 rs353291 polymorphism and specific rs353291 alleles. The distribution of the rs353291 C allele and CC genotypes was significantly higher in AS patients than in controls. Compared to individuals with the rs353291 TT genotype, carriers of the rs353291 C allele showed a significant increase in the plasma levels of sCD40L, IL-6 and MCP-1 and a decrease in miR-145 expression. After ox-LDL administration, the peripheral blood mononuclear cells (PBMCs) of rs353291 C allele carriers showed significantly increased CD40 and MCP-1 expression compared with those of individuals with the rs353291 TT genotype. These data suggest that the rs353291 C allele may increase susceptibility to atherosclerosis.

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Abbreviations: AS: atherosclerosis; PBMCs: peripheral blood mononuclear cells; ELISA: enzyme-linked immunosorbent assay; MCP-1: monocyte chemotactic protein 1; CAD: coronary artery disease; PAD: peripheral artery disease patients; SNPs: single nucleotide polymorphisms; sST2: soluble suppression of tumorigenicity 2; PPARγ: peroxisome proliferator-activated receptor-γ; LXRα: liver X receptor-α; PGC-1α: PPARγ co-activator-1α; ALDH2: aldehyde dehydrogenase2; miRNAs: microRNAs; 3’-UTR: 3’-untranslated region; ox-LDL: oxidized low-density lipoprotein; TNF-α: tumor necrosis factor-α; TGF-β: transforming growth factor-β; VSMCs: vascular smooth muscle cells; ECs: endothelial cells; BaPWV: brachial-ankle pulse wave velocity; PCR: polymerase chain reaction; OD: optical density; FCS: fetal calf serum; SDS-PAGE: SDS-polyacrylamide gel electrophoresis; PVDF: polyvinylidene difluoride; BMI: body mass index; SBP: systolic blood pressure; DBP: diastolic blood pressure; MALAT1: metastasis associated lung adenocarcinoma transcript 1.

Key Words: Atherosclerosis, miR-145, CD40, IL-6, MCP-1

Send correspondence to: Ren Guo, Department of Pharmacy, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China, Tel: 86-731-88618733, Fax: 86-731-88618455, E-mail: grfw2012@aliyun.com; Heng Yang, Department of Neurology, The Third Xiangya Hospital, Central South University, Changsha, Hunan 410013, P.R. China, Tel: 86-731-88618208, Fax: 86-731-88618208, E-mail:873008352@qq.com.