Journal of Molecular Medicine

, Volume 92, Issue 5, pp 473–485 | Cite as

Control of autophagy maturation by acid sphingomyelinase in mouse coronary arterial smooth muscle cells: protective role in atherosclerosis

  • Xiang Li
  • Ming Xu
  • Ashley L. Pitzer
  • Min Xia
  • Krishna M. Boini
  • Pin-Lan Li
  • Yang Zhang
Original Article
First Online:
Received:
Revised:
Accepted:

Abstract

Recent studies have indicated a protective role of autophagy in regulating vascular smooth muscle cells homeostasis in atherogenesis, but the mechanisms controlling autophagy, particularly autophagy maturation, are poorly understood. Here, we investigated whether acid sphingomyelinase (ASM)-regulated lysosome function is involved in autophagy maturation in coronary arterial smooth muscle cells (CASMCs) in the pathogenesis of atherosclerosis. In coronary arterial wall of ASM-deficient (Smpd1 −/− ) mice on Western diet, there were high expression levels of both LC3B, a robust marker of autophagosomes (APs), and p62, a selective autophagy substrate, compared with those in wild-type (Smpd1 +/+ ) mice. By Western blotting and flow cytometry, atherogenic stimulation of Smpd1 +/+ CASMCs with 7-ketocholesterol was found to significantly enhance LC3B expression and increase the content of both APs and autophagolysosomes (APLs). In Smpd1 −/− CASMCs, such 7-ketocholesterol-induced increases in LC3B and p62 expression and APs were further augmented, but APLs formation was abolished. Analysis of fluorescence resonance energy transfer between fluorescence-labeled LC3B and Lamp1 (lysosome marker) showed that 7-ketocholesterol markedly induced fusion of APs with lysosomes in Smpd1 +/+ CASMCs, which was abolished in Smpd1 −/− CASMCs. Moreover, 7-ketocholesterol-induced expression of cell dedifferentiation marker vimentin and proliferation was enhanced in Smpd1 −/− CASMCs compared with those in Smpd1 +/+ CASMCs. Lastly, overexpression of ASM further increased APLs formation in Smpd1 +/+ CASMCs and restored APLs formation in Smpd1 −/− CASMCs indicating that increased ASM expression is highly correlated with enhanced APLs formation. Taken together, our data suggest that the control of lysosome trafficking and fusion by ASM is essential to a normal autophagic flux in CASMCs, which implicates that the deficiency of ASM-mediated regulation of autophagy maturation may result in imbalance of arterial smooth muscle cell homeostasis and thus serve as an important atherogenic mechanism in coronary arteries.

Key messages

• Acid sphingomyelinase (ASM) controls autophagy maturation in smooth muscle cells.

• ASM maintains smooth muscle cell homeostasis and its contractile phenotype.

• ASM plays a protective role in smooth muscle dysfunction and atherosclerosis.

Keywords

Acid sphingomyelinase Autophagic flux Lysosome coronary arterial smooth muscle cell Coronary artery Atherosclerosis 
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Notes

Acknowledgments

This study was supported by grants from the National Institutes of Health (HL-57244, HL-075316, and HL-091464), National Institute of Health CTSA grant UL1TR000058, and VCU CCTR Endowment Fund.

Conflict of interest

None.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  • Xiang Li
    • 1
  • Ming Xu
    • 1
  • Ashley L. Pitzer
    • 1
  • Min Xia
    • 1
  • Krishna M. Boini
    • 1
  • Pin-Lan Li
    • 1
  • Yang Zhang
    • 1
  1. 1.Department of Pharmacology and Toxicology, School of MedicineVirginia Commonwealth UniversityRichmondUSA

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