Cyclophilin D induces necrotic core formation by mediating mitochondria-associated macrophage death in advanced atherosclerotic lesions

Atherosclerosis. 2024 Sep:396:118524. doi: 10.1016/j.atherosclerosis.2024.118524. Epub 2024 Jun 20.

Abstract

Background and aims: In advanced atherosclerotic lesions, macrophage deaths result in necrotic core formation and plaque vulnerability. Cyclophilin D (CypD) is a mitochondria-specific cyclophilin involved in the process of cell death after organ ischemia-reperfusion. However, the role of CypD in atherosclerosis, especially in necrotic core formation, is unknown. Therefore, this experiment aims to clarify the role of CypD in necrotic core formation.

Methods: To clarify the specific role of CypD, encoded by Ppif in mice, apolipoprotein-E/CypD-double knockout (Apoe-/-Ppif-/-) mice were generated. These mice were fed a high-fat diet containing 0.15 % cholesterol for 24 weeks to accelerate atherosclerotic lesion development.

Results: Deletion of CypD decreased the necrotic core size, accompanied by a reduction of macrophage apoptosis compared to control Apoe-/- mice. In RAW264.7 cells, siRNA-mediated knockdown of CypD attenuated the release of cytochrome c from the mitochondria to the cytosol induced by endoplasmic reticulum stress inducer thapsigargin. In addition, necroptosis, induced by TNF-α and caspase inhibitor, was attenuated by knockdown of CypD. Ly-6Chigh inflammatory monocytes in peripheral blood leukocytes and mRNA expression of Il1b in the aorta were decreased by deletion of CypD. In contrast, siRNA-mediated knockdown of CypD did not significantly decrease Il1b nor Ccl2 mRNA expression in RAW264.7 cells treated with LPS and IFN-γ, suggesting that inhibition of inflammation in vivo is likely due to decreased cell death in the atherosclerotic lesions rather than a direct action of CypD deletion on the macrophage.

Conclusions: These results indicate that CypD induces macrophage death and mediates necrotic core formation in advanced atherosclerotic lesions. CypD could be a novel therapeutic target for treating atherosclerotic vascular diseases.

Keywords: Apoptosis; Atherosclerosis; Cyclophilin D; Macrophages; Necrotic core.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Ly
  • Apolipoproteins E / deficiency
  • Apolipoproteins E / genetics
  • Apoptosis
  • Atherosclerosis* / genetics
  • Atherosclerosis* / metabolism
  • Atherosclerosis* / pathology
  • Cyclophilins / deficiency
  • Cyclophilins / genetics
  • Cyclophilins / metabolism
  • Diet, High-Fat
  • Disease Models, Animal
  • Interleukin-1beta / metabolism
  • Macrophages* / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Knockout, ApoE
  • Mitochondria* / metabolism
  • Mitochondria* / pathology
  • Necroptosis
  • Necrosis*
  • Peptidyl-Prolyl Isomerase F* / genetics
  • Peptidyl-Prolyl Isomerase F* / metabolism
  • Plaque, Atherosclerotic*
  • RAW 264.7 Cells

Substances

  • Peptidyl-Prolyl Isomerase F
  • PPIF protein, mouse
  • Apolipoproteins E
  • Cyclophilins
  • Interleukin-1beta
  • Apoe protein, mouse
  • Ly-6C antigen, mouse
  • Antigens, Ly