Cholesterol secoaldehyde induces apoptosis in H9c2 cardiomyoblasts through reactive oxygen species involving mitochondrial and death receptor pathways

K. Sathishkumar, Xueli Gao, Achuthan C. Raghavamenon, Narasimham Parinandi, William A. Pryor, Rao M. Uppu

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Cholesterol secoaldehyde (ChSeco), a putative product of the reaction of ozone with cholesterol in aqueous environments, has been shown to induce apoptosis in H9c2 cardiomyoblasts. This study further investigated the involvement of apoptotic-related proteins and gene expression using RT-PCR, Western blot, and appropriate biochemical assays. The RT-PCR analysis revealed that ChSeco activates the expression of genes involved in the death receptor (extrinsic) pathway. The significance of this pathway was also evident from the increased activity of caspase-8. The overexpression of Apaf-1, loss of mitochondrial transmembrane potential, release of cytochrome c, and increased activity of caspase-9 provide further evidence for the involvement of a mitochondrial (intrinsic) pathway. Time-course analysis of ChSeco-exposed H9c2 cells showed an upstream increase in the generation of reactive oxygen species (ROS) and an associated decrease in the intracellular glutathione. N-acetyl-L-cysteine and Trolox significantly attenuated the ChSeco-induced ROS formation and cytotoxicity and also down-regulated the expression of the genes of all the players in either pathway. This study clearly shows that ChSeco induces apoptosis in H9c2 cells through ROS generation and the activation of both the intrinsic and the extrinsic pathway.

Original languageEnglish (US)
Pages (from-to)548-558
Number of pages11
JournalFree Radical Biology and Medicine
Volume47
Issue number5
DOIs
StatePublished - Sep 1 2009
Externally publishedYes

Fingerprint

Death Domain Receptors
Reactive Oxygen Species
Apoptosis
Gene Expression
Genes
Polymerase Chain Reaction
Caspase 9
Caspase 8
Ozone
Acetylcysteine
Cytotoxicity
Cytochromes c
Gene expression
Membrane Potentials
Glutathione
Assays
Western Blotting
Chemical activation
Cholesterol
3-hydroxy-5-oxo-5,6-secocholestan-6-al

Keywords

  • Antioxidants
  • Apoptosis
  • Cholesterol secoaldehyde
  • Death receptor pathways
  • Free radicals
  • H9c2 cardiomyoblasts
  • Mitochondrial pathways
  • Oxysterols
  • Redox signaling

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

Cite this

Cholesterol secoaldehyde induces apoptosis in H9c2 cardiomyoblasts through reactive oxygen species involving mitochondrial and death receptor pathways. / Sathishkumar, K.; Gao, Xueli; Raghavamenon, Achuthan C.; Parinandi, Narasimham; Pryor, William A.; Uppu, Rao M.

In: Free Radical Biology and Medicine, Vol. 47, No. 5, 01.09.2009, p. 548-558.

Research output: Contribution to journalArticle

Sathishkumar, K. ; Gao, Xueli ; Raghavamenon, Achuthan C. ; Parinandi, Narasimham ; Pryor, William A. ; Uppu, Rao M. / Cholesterol secoaldehyde induces apoptosis in H9c2 cardiomyoblasts through reactive oxygen species involving mitochondrial and death receptor pathways. In: Free Radical Biology and Medicine. 2009 ; Vol. 47, No. 5. pp. 548-558.
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