Photodynamic therapy induces caspase-dependent apoptosis in rat CNV model.

Abstract

PURPOSE:

To investigate the mechanism of cell death in laser-induced choroidal neovascularization (CNV) after photodynamic therapy (PDT).

METHODS:

PDT was performed in Brown-Norway rats using laser light at a wavelength of 689 nm, irradiance of 600 mW/cm(2), and fluence of 25 J/cm(2) after intravenous injection of verteporfin at the doses of 3, 6, and 12 mg/m(2). Apoptotic cells in CNV were detected by TUNEL assay at 1, 3, 6, 15, 24, and 48 hours after PDT. Caspase activation at 1, 3, 6, 15, and 24 hours after PDT was determined by immunohistochemistry (IHC) with a cleaved caspase-3 or -9 antibody. Akt activity was determined by Western blot and IHC with a phosphorylated-Akt (pAkt) antibody. To investigate the roles of Akt in PDT-induced apoptosis, insulin-like growth factor (IGF)-1, an Akt activator, with or without wortmannin, an inhibitor of PI3K-Akt pathway, was injected into the vitreous before PDT.

RESULTS:

The number of TUNEL-positive cells in CNV increased at 3 hours after PDT and peaked at 6 hours, showing a dose dependence of verteporfin. Caspase activation was detected in TUNEL-positive cells. Dephosphorylation of Akt in CNV occurred within 1 hour. IGF-1 significantly activated Akt and suppressed the number of TUNEL-positive cells in CNV, and the effects of IGF-1 were diminished by wortmannin.

CONCLUSIONS:

PDT induced caspase-dependent apoptosis in CNV. These results suggest that PDT leads to dephosphorylation of Akt and subsequent activation of the caspase-dependent pathway. Understanding the intracellular signaling mechanisms of apoptosis in PDT may lead to more selective and effective treatment of CNV secondary to age-related macular degeneration.

• Authors: Akihisa Matsubara, Toru Nakazawa, Kosuke Noda, Haicheng She, Edward Connolly, Tara A Young, Yuichiro Ogura, Evangelos S Gragoudas, Joan W Miller
• Journal: Investigative ophthalmology & visual science (Invest Ophthalmol Vis Sci) Vol. 48 Issue 10 Pg. 4741-7 (Oct 2007) ISSN: 0146-0404 [Print] United States
• PMID: 17898299 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• Photosensitizing Agents
• Porphyrins
• Verteporfin
• Insulin-Like Growth Factor I
• Phosphatidylinositol 3-Kinases
• Proto-Oncogene Proteins c-akt
• Casp3 protein, rat
• Casp9 protein, rat
• Caspase 3
• Caspase 9

Topics

• Animals
• Apoptosis (drug effects)
• Blotting, Western
• Caspase 3 (metabolism)
• Caspase 9 (metabolism)
• Choroidal Neovascularization (drug therapy, enzymology, pathology)
• Disease Models, Animal
• Fluorescent Antibody Technique, Indirect
• In Situ Nick-End Labeling
• Insulin-Like Growth Factor I (pharmacology)
• Phosphatidylinositol 3-Kinases (metabolism)
• Phosphorylation
• Photochemotherapy
• Photosensitizing Agents (therapeutic use)
• Porphyrins (therapeutic use)
• Proto-Oncogene Proteins c-akt (metabolism)
• Rats
• Rats, Inbred BN
• Signal Transduction
• Verteporfin