Professional Documents
Culture Documents
Action of Antioxidants
J. A. Milner
Milnerj@mail.nih.gov
Includes:
.
hydroxyl radicals ( OH)
superoxide anions (O2-)
singlet oxygen (1O2)
hydrogen peroxides (H2O2)
organic peroxides (R-OOH)
nitric oxide
peroxynitrite
Generation of Reactive Oxygen
Species
Reticuloendothelium Peroxisome GSSG
O2 P450
oxidase
oxidase GSH
Glutathione Peroxidase
H2O
NADPH Cu,Zn O2
Catalase
oxidase Superoxide
Dismutase H2O2 Thioredoxin reductase
O2- O2 -
O2 Fe+2
Xanthine oxidase /Cu+
1O Fe+3 .OH
O2 2 /Cu+2
O2 O2- H2O2 H
O2
2O
Mitochondria
Nitric Oxide Dependent Reactions
Peroxynitrite
O2 H202 •OH
•Damage DNA,
RNA
•Oxidize Proteins
(enzymes, histones)
Non-essential
glutathione, small peptides
host of phytochemicals (thousands in food
supply)
In Vitro Measures of Antioxidant
Capacity
• ABTS Assay for Antioxidant Activity (Miller et al. 1997).
- .
O2 , OH , 1O2 8-isoprostane
Mitochondrion Vitamin E
tocopherol a free radical (peroxyl) scavenger within
membranes
-TH + LOO ° Æ -T ° +LOOH
-T ° + ascorbate Æ -TH + ascorbate°
Effects of Dietary Supplements on
Oxidative Damage Markers
• In several studies, vitamin E and diet supplement
mixtures have been shown to favorably influence
indicators of oxidative status, such as
susceptibility of LDL to oxidation
• Randomized clinical trials have generally not
confirmed a beneficial effect of supplements (e.g.,
vitamin E) on disease risk or outcome (Blumberg
Nutr Clin Care 2002;5:50-5 )
α-tocopheryloxybutyric Acid (TE) and
Cell Apoptosis (% Control)
700
600
500
Percent
400
300 Control
TS
200
TE
100
0
Viability DNA Caspase 3
Fragmentation Activity
TE = modified vitamin E with no antioxidant properties
Akazawa 2002 Jpn J Pharm 89: 417.
Arachidonic Acid Metabolic Pathway
Linoleic Acid
Arachidonic Acid Ceramide Apoptosis
Lipoxygenases Cyclooxygenases
Dietary
Prostaglandins
HETEs
Antioxidants (PGG2)
PPAR Leukotrienes Fatty Acids
PPAR
Vitamin E Placebo
7 + Years Treatment
Endpoint Prostate Cancer Incidence
The TRAMP Mouse
• Transgenic Adenocarcinoma of Mouse Prostate (TRAMP)
animal model that expresses the oncogene SV40 T antigen
specifically in the epithelium of the prostate.
PLA2
β-Catenin TEA Map
Arachidonic Acid TEA
Kinases
β-Catenin Tcf
COX-2 AP-1
PGE2
TEA
c-Myc
Proliferation
Apoptosis
Wargovich Tumorigenesis
(personal communication)
Candidate Genes Responsive to (-)
Epigallocatechin-3-Gallate in Human Prostate
Cancer (LNCaP) Cells
6-
Antioxidant
reduces
% Apoptosis +
efficacy of
4- cisplatin
2- **
*
0-
Control E CP CP+E
15 - 3-
Tumor Volume
% Apoptosis +
10 - 2-
**
5- 1-
0- 0-
Control Poor Control Poor
Salganik, et al. Carcinogenesis 21: 909, 2000.
One Size Does Not Fit All!