Regulatory role of IKKɑ in myocardial ischemia/reperfusion injury by the determination of M1 versus M2 polarization of macrophages

J Mol Cell Cardiol. 2018 Oct:123:1-12. doi: 10.1016/j.yjmcc.2018.08.021. Epub 2018 Aug 25.

Abstract

The IκB kinase (IKK) complex plays a well-documented role in cancer and immune system. This function has been widely attributed to its role as the master regulator of the NF-κB family. Particularly, IKKɑ, a member of IKK complex, is reported to have various regulating effects in inflammatory and malignant diseases. However, its role as well as its mechanism of function in macrophages following myocardial ischemia and reperfusion (I/R) injury remains unexplored. In vivo, sham or I/R operations were performed on macrophage-specific IKKɑ knockout (mIKKɑ-/-) mice and their IKKɑflox/flox littermates. We ligated the left anterior descending (LAD) coronary artery of I/R groups simulating ischemia for 30 min, followed by a reperfusion period of 3 days and 7 days, respectively. The hearts of mIKKɑ-/- mice exhibited significantly increased inflammation and macrophage aggregation as compared to their IKKɑflox/flox littermates. Moreover, in the mIKKɑ-/- group subjected to I/R macrophages had a tendency to polarize to M1 phenotype. In vitro, we stimulated RAW264.7 cells with Lipopolysaccharides (LPS) after infection by the lentivirus, either knocking-down or overexpressing IKKɑ. We discovered that a deficiency of IKKɑ in RAW264.7 caused increased expression of pro-inflammatory markers compared to normal RAW264.7 after LPS stimulation. Inversely, pro-inflammatory factors were inhibited with IKKɑ overexpression. Mechanistically, IKKɑ directly combined with RelB to regulate macrophage polarization. Furthermore, IKKɑ regulated MEK1/2-ERK1/2 and downstream p65 signaling cascades after LPS stimulation. Overall, our data reveals that IKKɑ is a novel mediator protecting against the development of myocardial I/R injury via negative regulation of macrophage polarization to M1 phenotype. Thus, IKKɑ may serve as a valuable therapeutic target for the treatment of myocardial I/R injury.

Keywords: IKKɑ; MEK1/2/ERK1/2; Macrophages polarization; Myocardial ischemia and reperfusion injury; NF-κB.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cytokines / metabolism
  • Disease Models, Animal
  • Echocardiography
  • Gene Expression
  • I-kappa B Kinase / genetics
  • I-kappa B Kinase / metabolism*
  • Immunohistochemistry
  • Immunophenotyping
  • Inflammation Mediators / metabolism
  • Macrophage Activation / immunology*
  • Macrophages / immunology*
  • Macrophages / metabolism*
  • Mice
  • Mice, Knockout
  • Mice, Transgenic
  • Mitogen-Activated Protein Kinases / metabolism
  • Myocardial Reperfusion Injury / diagnosis
  • Myocardial Reperfusion Injury / etiology*
  • Myocardial Reperfusion Injury / metabolism*
  • Myocardium / metabolism
  • NF-kappa B / metabolism
  • RAW 264.7 Cells
  • Signal Transduction

Substances

  • Cytokines
  • Inflammation Mediators
  • NF-kappa B
  • CHUK protein, human
  • I-kappa B Kinase
  • Mitogen-Activated Protein Kinases