PMID- 10102288
OWN - NLM
STAT- MEDLINE
DCOM- 19990402
LR  - 20220318
IS  - 0146-0404 (Print)
IS  - 0146-0404 (Linking)
VI  - 40
IP  - 5
DP  - 1999 Apr
TI  - Reactive oxygen species-induced apoptosis and necrosis in bovine corneal 
      endothelial cells.
PG  - 911-9
AB  - PURPOSE: The loss of corneal endothelial cells associated with aging and possibly 
      other causes has been speculated to be related to exposure to reactive oxygen 
      species (ROS). The current study was conducted to investigate, by use of 
      photosensitizers, the underlying mechanisms involved in the death of bovine 
      corneal endothelial cells (BCENs) caused by ROS. METHODS: BCEN cells in primary 
      culture were treated with a photosensitizer (riboflavin or rose bengal) with 
      light exposure. The patterns of cell damage and death were assessed using an 
      acridine orange-ethidium bromide differential staining method, TdT-mediated dUTP 
      nick-end labeling (TUNEL) assay, and transmission electron microscopy. The 
      cytotoxicity was assayed by mitochondrial function using 
      3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) testing. 
      Antioxidants, including catalase, L-histidine, salicylic acid, and superoxide 
      dismutase, were used to determine the types of ROS involved. Activation of 
      nuclear factor (NF)-kappaB was examined by fluorescent immunocytochemistry with 
      anti-p65 antibody. RESULTS: Light-irradiated riboflavin or rose bengal resulted 
      in a significant decrease in viability of BCEN cells. Chromosomal condensation 
      and fragmentation were observed in apoptotic cells, and membrane lysis and damage 
      of cell ultrastructures were observed in necrotic cells. Riboflavin induced 
      apoptosis at 30 minutes and thereafter and induced necrosis after 2 hours. Rose 
      bengal was shown to cause similar effects within half the time required for the 
      effects of riboflavin. Catalase and salicylic acid were found to provide 
      protection for BCENs from cytotoxic effects of riboflavin, and L-histidine was 
      found to protect BCENs from cytotoxicity induced by rose bengal. Kinetic studies 
      using immunocytochemistry showed that NF-kappaB was translocated into the nucleus 
      within 15 minutes and 30 minutes after treatment with rose bengal and riboflavin, 
      respectively. CONCLUSIONS: The cytotoxic effects of photo-irradiated riboflavin 
      and rose bengal are shown to be mediated by two distinct but parallel pathways, 
      one leading to apoptosis and the other to necrosis. Possible involvement of 
      NF-kappaB in cell death is suggested. These findings provide potential leads for 
      future investigation into the molecular mechanisms of loss of corneal endothelial 
      cells related to aging, oxidative stress, and possibly other similar causes.
FAU - Cho, K S
AU  - Cho KS
AD  - Department of Ophthalmology and Visual Science, College of Medicine, The Catholic 
      University of Korea, and Catholic Research Institutes of Medical Science, Seoul.
FAU - Lee, E H
AU  - Lee EH
FAU - Choi, J S
AU  - Choi JS
FAU - Joo, C K
AU  - Joo CK
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Invest Ophthalmol Vis Sci
JT  - Investigative ophthalmology & visual science
JID - 7703701
RN  - 0 (Photosensitizing Agents)
RN  - 0 (Reactive Oxygen Species)
RN  - 0 (Tetrazolium Salts)
RN  - 0 (Thiazoles)
RN  - 1ZPG1ELY14 (Rose Bengal)
RN  - EN464416SI (Ethidium)
RN  - EUY85H477I (thiazolyl blue)
RN  - F30N4O6XVV (Acridine Orange)
RN  - TLM2976OFR (Riboflavin)
SB  - IM
MH  - Acridine Orange
MH  - Animals
MH  - *Apoptosis
MH  - Cattle
MH  - Cell Survival/drug effects
MH  - Cells, Cultured
MH  - Endothelium, Corneal/metabolism/*pathology
MH  - Ethidium
MH  - In Situ Nick-End Labeling
MH  - Mitochondria/drug effects
MH  - Necrosis
MH  - Photosensitizing Agents/pharmacology
MH  - *Reactive Oxygen Species
MH  - Riboflavin/pharmacology
MH  - Rose Bengal/pharmacology
MH  - Tetrazolium Salts
MH  - Thiazoles
EDAT- 1999/04/02 00:00
MHDA- 1999/04/02 00:01
CRDT- 1999/04/02 00:00
PHST- 1999/04/02 00:00 [pubmed]
PHST- 1999/04/02 00:01 [medline]
PHST- 1999/04/02 00:00 [entrez]
PST - ppublish
SO  - Invest Ophthalmol Vis Sci. 1999 Apr;40(5):911-9.