NR4A2 alleviates cardiomyocyte loss and myocardial injury in rats by transcriptionally suppressing CCR5 and inducing M2 polarization of macrophages

Microvasc Res. 2022 Mar:140:104279. doi: 10.1016/j.mvr.2021.104279. Epub 2021 Nov 10.

Abstract

Background: CC chemokine receptor 5 (CCR5) has been demonstrated to be correlated to activation of pro-inflammatory immune cells and tissue injury. This study focused on the role of CCR5 in myocardial injury in rats with diabetic cardiomyopathy (DCM) and the mechanism of action.

Methods: A rat model of DCM was induced by streptozotocin (STZ). CCR5 was knocked down in rats to determine its role in myocardial injury and immune cell infiltration. The upstream regulators of CCR5 were bioinformatically predicted and the binding between nuclear receptor subfamily 4 group A member 2 (NR4A2) and CCR5 was validated. The portion of M1 and M2 macrophages in tissues was determined by flow cytometry or double-labeling immunofluorescence. Rat bone marrow mononuclear cells (BMMCs) were treated with granulocyte/macrophage colony stimulating factor (GM-CSF/M-CSF) and co-cultured with H9C2 cells for in vitro experiments.

Results: STZ-treated rats had impaired cardiac function and increased levels of creatine kinase-MB, cardiac troponin I and lactate dehydrogenase. CCR5 inhibition significantly alleviated myocardial injury in rats and reduced the portion of M1 macrophages in rat cardiac tissues. NR4A2, which could suppress CCR5 transcription, was poorly expressed in rats with DCM. NR4A2 overexpression played a similar myocardium-protective role in rats. In vitro, overexpression of NR4A2 induced M2 polarization of macrophages, which protected the co-cultured H9C2 cells from high glucose-induced damage, but the protective role was blocked after CCR5 overexpression.

Conclusion: This study demonstrated that NR4A2 suppresses CCR5 expression and promotes M2 polarization of macrophages to alleviate cardiomyocyte loss and myocardial injury.

Keywords: CCR5; Diabetic cardiomyopathy; Immune cell infiltration; Macrophages; NR4A2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Coculture Techniques
  • Diabetic Cardiomyopathies* / genetics
  • Diabetic Cardiomyopathies* / immunology
  • Diabetic Cardiomyopathies* / metabolism
  • Diabetic Cardiomyopathies* / pathology
  • Disease Models, Animal
  • Down-Regulation
  • Macrophages* / immunology
  • Macrophages* / metabolism
  • Macrophages* / pathology
  • Male
  • Myocytes, Cardiac* / immunology
  • Myocytes, Cardiac* / metabolism
  • Myocytes, Cardiac* / pathology
  • Nuclear Receptor Subfamily 4, Group A, Member 2* / genetics
  • Nuclear Receptor Subfamily 4, Group A, Member 2* / metabolism
  • Phenotype
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, CCR5* / genetics
  • Receptors, CCR5* / metabolism
  • Signal Transduction
  • Transcription, Genetic*

Substances

  • Nr4a2 protein, rat
  • Nuclear Receptor Subfamily 4, Group A, Member 2
  • Receptors, CCR5
  • CCR5 protein, rat