Ελευθερες ριζες-στρες PDF

- -
:
: ,
ISBN: 978-960-603-417-6
Copyright , 2015
Creative Commons -
- 3.0.
https://creativecommons.org/licenses/by-nc-sa/3.0/gr/

9, 15780
www.kallipos.gr
,
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1 - : ................................................... 15
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: ............................................................. 23
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(2.-) .................................................................................................................................. 28
(H2O2) ............................................................................................................................ 30
(.) ....................................................................................................................... 30
- - (ROO. R.) ................................................................................................................... 31
(HOCl) ......................................................................................................................................... 32
(singlet O2, 1O2)............................................................................................................................... 33
(3)............................................................................................................................................................................ 33
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1 ................................................................................................................................................... 35
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2 ................................................................................................................................................... 36
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3 - ................................... 37
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1 ................................................................................................................................................... 47
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2 ................................................................................................................................................... 47
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4 - in vivo ............................................ 48
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2 ................................................................................................................................................... 60
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5 - :
........................................................................................................................................... 61
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2.- .................................................................................................................... 62
Fenton ............................................................................................................................................................. 63
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DNA............................................................................................................................................ 69
DNA ...................................................................................................................................... 71
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3 ................................................................................................................................................... 83
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6 - :
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(GPx) ....................................................................................................................... 90
GSSG NADP+ ........................................................................................................ 91
(Prx) ...................................................................................................................................................... 93
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............ 97
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NAD(P)H: 1 (NQO1, DT-) ................................................................................ 101
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7 - :
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8 - :
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(labile iron) .................................................................................................................... 137
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2 ................................................................................................................................................. 149
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9 - (Redox
Signaling) .......................................................................................................................................... 150
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Keap1/Nrf2 ............................................................................................................................. 173
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-
.

1
O2
AREs antioxidant response elements
Arg
Asc
Asc.
ASK apoptosis signal-regulating kinase
ASK1 apoptosis signal-regulating kinase 1
AT
BH4
CAT
CGD chronic granulomatous desease
CH2=CH2
-CH2-S-
-CH2-SH
-CH2-SO3H
-CH2-SOH
-CH2-S2H
CH3(CH2)3CH3
CH3-CH3
Cl.
CoQ Q
CoQH. Q
CP
CuZnSOD ,
C32-
DCF --
Dcytb
DFO
DFP
DHAsc
EDRF endothelial derived relaxing factor
EGF epidermal growth factor
EGF
eNOS endothelial NOS
EPR electron paramagnetic resonance
EpREs electrophile response elements
FAD -
Fe+2-2
Fe+3
FeSO4
FeSOD
G6P 6-
G6P-DH 6-
GFP green fluorescence protein
GPx
GR glutathione reductase
GR
GS.
GSH
GSH-S-transferase
GSNO
GSSG
GST
H 2 2
HETE
Hjv
HO heme oxygenase
HO1 1
HO2.
HOCl
HOO-
HPETE
iNOS inducible NOS
IREs iron responsive elements
IRIDA iron refractory iron deficiency anemia
IRPs iron-regulatory proteins
k
LDL
LIP labile iron pool
LIP labile iron pool
LO.
LOO.
LOOH
LO.
LPS
LTA4
MAO
MHC major histocompatibility complex
MnSOD
MPO
Msr
NBT nitroblue tetrazolium
nNOS neuronal NOS
NO
NO2
NOS NO-synthase
NQO1
Nrf2 Nuclear factor - erythroid-2 related factor 2
NTBI non transferrin bound iron
N3-
O2.-
O3
OCl-/Cl-
OH-
OH.
ONOO-
OSCN-
PARP (ADP-)
PC1 Peroxy Crimpson 1
PDEs
PDGF platelet derived growth factor
PDI
PG
PG1 Peroxy Green 1
PGHS
PG2
PKA protein kinase A
Prx
PSSG
-R.
R-CO.
RO.
ROO.
ROOH
ROS reactive oxygen species
RS.
RSNO
-S.
SCN-
Se
-SH
-SO2H
SOD
SOO. -
-S-O-O.
Srx
TfR
TLR toll-like receptors
Trx
TrxR thioredoxin reductase
TrxR
TX
Ur.
XDH
YFR yellow fluorescence protein
Zn
-Toc. -
-Toc -

, :
.

.
1950, (free
radicals) ,
[Gerschman et al., 1954] [Herman Harman, 1956],
.

(DNA, , ).
1969, Irvin Fridovich Joe McCord ,
, ,
(2.-).
(SOD).
, ,

,
.
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,
,
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.

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, 2015
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.
, ,
. (,
) in vitro,
in vivo. , ,
.

, .

, ..
, , ..

. , , , ,
, ,
,
(Halliwell and Gutteridge, 2007).
100 ,
,
. , 1940 1950, 10.000
, , (retrolental fibroplasia).
20
.
(free radical theory of aging)
1950 (Gerschman, Gilbert et al., 1954; Harman, 1956),

.
(SOD)
McCord Fridovich 1969 ( 1) , ,
.-
2
(McCord and Fridovich, 1969).

, . , ,
(
, EPR ESR), .

( 100), ..
, ,
,
, , ,
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,
, .
,
, ,
(redox signaling).
. ,
,
.

, . , ,

(Galaris and Pantopoulos, 2008).

, (Oxidative Stress) (Sies, 2015).
(, , ,
...) , , ,
.

,
, ,
(Jones and Sies, 2015).
;
, ,
- -
, .. (C32 ) (N3 ). ,
, : (free radical)
, - ,
(spin).
, ,
, .
*
,
( 1).
1:
-.
() , ()
. , 2
, 2.
2: .
* :
: .
: .
:
.
: .
- .
, ( ) ( )
,
. 10 -7 M
pH=7,
.

,
. ,
(22)
( 2 3):
,
,
. .
, . ( ) (Cl )
( 3) ,
(diffusion controlled reactions).
3: .
, , , -
-, , ,
.
( ) (2.-),
- , , ,
. , ,
.
, ,
,
.

H.J.H. Fenton 1894 (Fenton, 1894).
,
(FeSO4) (22)
. Fenton
. , ,
, Fenton.
( 4)
(.) 22,
.

[Fe.OH]3+ [Fe=O]2+

(Merkofer, Kissner et al., 2006).
--
(Gomberg, 1900) ( 4).
4: - -- .
Gomberg
( 4). Fenton, Gomberg
, . ,
.

.
( 5).
5: .
,
- ,
( ).
, .
, , ,
.
( 6).
6: .
() ,
, ()
(.) Q, (Q.-). , ,
Q
. () ,
. . , ,
. ()
.
Q C ( ),
, (-)

(, LDL ...). , ,
, .
,
.

, ,
. , ,
,
. , ,

. ( 10) ,
,
(Barbouti, Briasoulis et al., 2010;
Kitsati, Fokas et al., 2012; Melidou, Riganakos et al., 2005; Nousis, Doulias et al., 2005). ,
, .
, ,
,
.
,
. ,
.
Barbouti, A., Briasoulis, E. and Galaris, D. 2010. Protective Effects of Olive Oil Components
Against Hydrogen Peroxide-Induced DNA Damage: The Potential Role of Iron Chelation. In V. R.
Preedy and R. R. Watson eds. Olives and Olive Oil in Health and Disease Prevention. Oxford,
Academic Press. pp. 1103-1109.
Fenton, H.J.H. 1894. LXXIII.-Oxidation of tartaric acid in presence of iron. Journal of the Chemical
Society, Transactions 65(0) 899-910.
Galaris, D. and Pantopoulos, K. 2008. Oxidative stress and iron homeostasis: mechanistic and health
aspects. Crit Rev Clin Lab Sci 45(1) 1-23.
Gerschman, R., Gilbert, D.L., Nye, S.W., Dwyer, P. and Fenn, W.O. 1954. Oxygen poisoning and x-
irradiation: a mechanism in common. Science 119(3097) 623-626.
Gomberg, M. 1900. An instance of trivalent carbon: triphenylmethyl. Journal of the American

Chemical Society 22(11) 757-771.
Halliwell, B. and Gutteridge, J. 2007. Free Radicals in Biology and Medicine. Fourth ed. Oxford
U.K.: Oxford University Press.
Harman, D. 1956. Aging: a theory based on free radical and radiation chemistry. J Gerontol 11(3)
298-300.
Jones, D.P. and Sies, H. 2015. The Redox Code. Antioxid Redox Signal 23(9) 734-746.
Kitsati, N., Fokas, D., Ouzouni, M.D., Mantzaris, M.D., Barbouti, A. and Galaris, D. 2012.
Lipophilic caffeic acid derivatives protect cells against H2O2-Induced DNA damage by chelating
intracellular labile iron. J Agric Food Chem 60(32) 7873-7879.
McCord, J.M. and Fridovich, I. 1969. Superoxide dismutase. An enzymic function for erythrocuprein
(hemocuprein). J Biol Chem 244(22) 6049-6055.
Melidou, M., Riganakos, K. and Galaris, D. 2005. Protection against nuclear DNA damage offered
by flavonoids in cells exposed to hydrogen peroxide: the role of iron chelation. Free Radic Biol Med
39(12) 1591-1600.
Merkofer, M., Kissner, R., Hider, R.C., Brunk, U.T. and Koppenol, W.H. 2006. Fenton chemistry
and iron chelation under physiologically relevant conditions: Electrochemistry and kinetics. Chem
Res Toxicol 19(10) 1263-1269.
Nousis, L., Doulias, P.T., Aligiannis, N., Bazios, D., Agalias, A., Galaris, D. and Mitakou, S. 2005.
DNA protecting and genotoxic effects of olive oil related components in cells exposed to hydrogen
peroxide. Free Radic Res 39(7) 787-795.
Sies, H. 2015. Oxidative stress: a concept in redox biology and medicine. Redox Biol 4 180-183.
1

;

, - ,
(spin).
2
;
,
:
() ,
. .
() (.) Q
. , (Q.+).
, , Q
.
() .
, , (
.
), .
()
.
Q C ( ),
, (-),

(, LDL ...).
2 -

,
, ,
.
,
,
, . 2
. , ,
2,
.

, .

(O2).
2.5 ,
, ( , ),
2
. , ,
, 2 (Halliwell and Gutteridge, 2007).
O2 (O3) .
,
.
O2 ~21%
(78%).
2 , ,
, 2.

,
2. ,
2
.
,

ATP , .
:
, , ,
, NAD+
FAD ( 1).
NAD+ FAD ,
ATP.
,
( 1).

(NADH FADH2) -
( -), Q
() , 2. 2
, ( 2)
.
2
( IV) . ,
4 C (
Fe2+ Fe3+) 2 ( 1):
(2 2
Cu ),
2, 2.
,

.
( ).
( V), ATP
ADP ATP.
85 90% 2.
15% (.., d- ,
..) (.., , ),
( P-450).
1: . ()
() (-) Krebs. ()
ATP .

,
. ,

. 1954, Gershman Gilbert,
,
2
2 (Gerschman, Gilbert et al., 1954). ,
, 2
-
2. , 2 .
-
( Krebs), 2.
( -, 4Fe4S). ,
2,
2. , ,

.

2 ( 0.2 2%)
, , ,
2.
2:
.

.
2 , ,
2.- ,
.
2
( 2-6):
, 2.-, (2.), 22
(.)
.
,
,
1,
. 1
2
. .
, , ,
.

2.-
2.
.

R.
RO.
2. .
-
22
ROOH
1
O2
3
HOCl
-
1.

, ,
.
(spin) . ,
2, . , ,
Lewis
, .
(triplet 2)
( 3) 2.

( ) , Pauli,
. ,
( 2 singlet 2),
(1g+O2 ) (1g O2 ),
. , , ,
Pauli.
2
. ,
2
.
3:
.
triplet 2 singlet 2,
2 2 ,
. M (multiplicity) = 2S + 1 ( S
). , 2 S = 2x1/2= 1, = 3 (triplet),
S = -1/2 + 1/2 = 0 = 1 (singlet).
(2.-)
2
( 2 3), .
(.)
, (+) (e-) .
(2.), pH
2.- + ( 7 8):
RH .
pK O2. 4.8,
pH,
( )
. 2., 2.-,
, .
+ 2.-
(2.).
2.-
Fe, -.
, ,
. ,
( 4Fe-4S)
2.-.
,
2.- 2. ( 9-12).
k1 < 1.0 x 102, k2 = 7.6 x 105 k3 = 8.2 x 107 M-1s-1. H

12 1.9 x 109 M-1s-1
2.- 2. .
2 9-12, e-
O2.- 22 O2.-. ,
O2.-, H2O2.
2.-, , . ,
C ( 13):
2.-
(SOD). , 2.-
.
,
. ( 3).
(H2O2)
22 2.-, , ()
, D-
, p66hc ,
H2O2 e- O2.
10-7-10-8 (Sies, 2014).
, H2O2
O2 (Winterbourn, 2013). ,
, (-SH), Fe-S, ..
, ,
, ()
.

, , . ,
, DNA, , 22
. 22
( Fe),
(Barbouti,
Doulias et al., 2001; Kitsati, Fokas et al., 2012; Melidou, Riganakos et al., 2005). ,
, 22
, ,
( +4 )
.

(Galaris and Korantzopoulos, 1997).
.
22
( ) (second messenger),
9 (Truong and Carroll, 2012; Winterbourn, 2013).
(.)
120 1894 H.J.H. Fenton
22 ,
( Fenton) (Fenton, 1894). ,
,
(Merkofer, Kissner et al., 2006).
, (.) (
14):
, , .
, 22,
. ( 15):
, ,
. ,
DNA, , ,
.
:
) , ) )
.
-
, .
, ,
. , ,
.
- - (ROO. R.)
- - (ROO. RO., )
.
(.., .) ,
. , , 2
- ( 16):
ROO.
(ROOH).
( ), ROO. RO. ( 17 18):

, .
, ,
.
. , ROO
, ( Russell),
( 19):
, - -
, .
, ,
.
(HOCl)
(HOCl) in vivo (MPO),
, .
22 ( 20):
-
p, HOCl 50:50 HOCl OCl ,

. HOCl
, 1- ,
(.., ).
, HOCl 1-,
,
, , .
,
, HOCl
. HOCl , ,
DNA ( ), .
in vivo ,
, HOCl.
, (.. GSH), NAD(P)H ..
HOCl
2.- Fe2+ ( 21 22),
( 23):
, ,
.
,
. ,

DNA. , ( )
.
, HOCl, 22,

.
(singlet O2, 1O2)

, ( 2). 1g+
, , 1g ,
. 1g ,
, ,
. . ,
,
. , , 2,
1
2, .
, FAD FMN, ,
, , . 12 ,
, ,
.
.
- -. (
retina), , ,
12 .
12 , , , ,
, .
, ,
.
12 ,
. 12
(--)
( 24):
, S, 12
, , .
(3)

.
. , ,
, ,
. , ,

, .
,
, ( 4),
.
4: .

, . , ,
, ,
, ,
.
2 NADH NADPH, ,
, , . 12 , ,
, . ,
, ,
, , ,
.., .
Barbouti, A., Doulias, P.T., Zhu, B.Z., Frei, B. and Galaris, D. 2001. Intracellular iron, but not
copper, plays a critical role in hydrogen peroxide-induced DNA damage. Free Radic Biol Med 31(4)
490-498.
Fenton, H.J.H. 1894. LXXIII.-Oxidation of tartaric acid in presence of iron. Journal of the Chemical
Society, Transactions 65(0) 899-910.
Galaris, D. and Korantzopoulos, P. 1997. On the molecular mechanism of metmyoglobin-catalyzed

reduction of hydrogen peroxide by ascorbate. Free Radic Biol Med 22(4) 657-667.
39(12) 1591-1600.
Merkofer, M., Kissner, R., Hider, R.C., Brunk, U.T. and Koppenol, W.H. 2006. Fenton chemistry
and iron chelation under physiologically relevant conditions: Electrochemistry and kinetics. Chem
Res Toxicol 19(10) 1263-1269.
Sies, H. 2014. Role of metabolic H2O2 generation: redox signaling and oxidative stress. J Biol Chem
289(13) 8735-8741.
Truong, T.H. and Carroll, K.S. 2012. Redox regulation of epidermal growth factor receptor signaling
through cysteine oxidation. Biochemistry 51(50) 9954-9965.
Winterbourn, C.C. 2013. The biological chemistry of hydrogen peroxide. Methods Enzymol 528 3-
25.
1
, .
;
2
( IV) . 4
C ( Fe2+ Fe3+)
2 :
2, ,
, , ATP,
.
, ,

, .
2
(MPO) ,
. ;
;
22
(HOCl):
HOCl
. , ,
, 1- ,
(.. ).
HOCl 1-,

, , .
,
, HOCl
.
, HOCl DNA (
) .
HOCl in vivo
, ,
HOCl.
, HOCl
2.-, Fe2+, .

. ,

. ,
DNA. ,
( )
.
, HOCl, 22,

.
3 -

(), ,
. 1980, ,
, , ,
, .
NO (NOS):
(eNOS), (nNOS)
(iNOS),
. , , . ,
,
-.
GTP, cGMP.
(.-). , (-
), , ,
,
.

, .

(NO., NO2., N2O)
, ,
. , ,
1980
(EDRF, endothelial derived relaxing factor) ().

Ascanio Sobrero, 1847. Sobrero
( )
( ).
. ,

( Marsh and Marsh, 2000).
EDRF
(Gryglewski, Palmer et al., 1986; Ignarro, Buga
et al., 1987). ,
,
(Moncada, Palmer et al., 1991; Palmer, Ashton et al., 1988).

1990. ,
(NOS, NO synthases) (Bredt, Hwang et al., 1991),
-
( ) (Beckman, Beckman et al., 1990), -

(Stamler, Singel et al., 1992),
(Brown and Cooper, 1994).
- .
, 2 ,
, ,
in vivo. ,
,
(). , ,

(/).
() , , , ,
,
. , 15 ,
. ,
( 1). ,
.
, , ,
(Beckman and Koppenol, 1996).
1: 2, 2.
,
1.
, 2,
, .
,
, , ,
, ,
(Martinez-Ruiz, Cadenas et al., 2011).
(k) NO ,

( 2):
) , GTP
GMP (cGMP).
,
.
) -,
.
-
) 2.-, ( ),
(Szabo, Ischiropoulos et al., 2007).
2: ,
. : (i)
(), (ii) , (),
(iii) , cGMP
().
, ,
. ,
,
,
in vivo. , 2 ( 1),
, in vivo,
2 , (Beckman and Koppenol, 1996).
in vivo
,
in vivo.
(S, NO-synthase)
(Arg) ( 3).
3: (A)
. -
. (B) NOS, G, NG--
(ADMA) G- (NMMA).
, , NADPH O2
.
: (eNOS, endothelial NOS),
(nNOS, neuronal NOS), (iNOS,
inducible NOS), (
) .
Ca2+, iNOS
Ca2+,
Ca2+ .
NOS 4.
4: NOS.
NADPH, FAD FMN , ,
, .
, NOS ,
NADPH, FAD FMN,
NADPH .
Ca2+
.
, , ( Fe3+ Fe2+),
2. , ,
2, . , ,
(4).
Ca2+
, eNOS nNOS.

. iNOS,
, , Ca2+
.
,
,
( Fe4+ ),
(Stuehr, 2004).
, (LPS) (INF-,
TNF-, IL-1 ), ,
iNOS . , , ,
, , NF- (
, -- ), .. ,
, iNOS ,
.
NOS , ,
in vivo
. G, NG-- (, ,
, ADMA) G- (NMMA) ( 3).
( D)
(Tsikas, Boger et al., 2000).
N
,
( ), ,
. ,
.
,
.- -
(2 ), ( )
(Pryor and Squadrito, 1995).
,
, .

GMP,
-GMP (cGMP) GTP ( 2).
cGMP
,
.
cGMP
(PDEs).
cGMP . ,
, PDE5
.

(Wong, Whelan et al., 2006).
, , ,
, , . ,

,
.
eNOS nNOS
, (Nathan and Xie, 1994). ,
, iNOS
,
.
( 5).
5:
.
-

, ,
.-
, 2 .
, (diffusion controlled). T
-
( ), 2.- ( 2)
.
6.7 x 109 M-1s-1. , ,

SOD ( 6).
6: 2.- , SOD
.
, 2.-
. SOD
M 2.-.
, , .. ,
5 10 nM 2.- SOD. , ,
M, SOD
-
. 2.- :
2.- ,
(.., ) 2.-.
(2) , 3.
in vivo
.
.
, .

, .
S-
S- -
- (R-S-N=O).
- .
,
. ,
, S- (Martinez-Ruiz,
Cadenas et al., 2011).
( ) - .

(site specificity)
. ,
22 (redox signaling).
(RSNO) <1 M
.
,
, ,
4:
, ,
in vivo
(Doulias, Greene et al., 2010). ,
S-
- (Doulias, Tenopoulou et al., 2013).

,
( 7).
3- -
(Ischiropoulos, 1998).
,
.
3- .
3-
.
7: S- 3-.
.
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1

. ;
.
NOS
. (L-, NADPH,
) , L-. ,
NADPH, FAD FMN , ,
.
,
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, . eNOS

. eNOS nNOS Ca2+
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.
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;
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.
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( Halliwell and Gutteridge, 2007).

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in vivo .
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. ,
.
.
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(~100 )
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NADPH ( 1).
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TLR
. p47 p67,
p22 gp91,
.
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. ,
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.
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, NADPH ,
(Babior, 2000).
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, , .
, ,
, .
, (LPS) ,
, RNA DNA.
TLR
( ), , ,
p47 p67
NADPH (Babior, 1999). ,

, ( 1).
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, (respiratory burst).
(oxidative
burst), ,
.
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NADPH gp91phox
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, 2.- 22
, ,
22.
, , ,
, ().
22 (HOCl) ( 1)
(Hampton, Kettle et al., 1998):
,
, .
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(CGD, Chronic Granulomatous Disease)
NADPH .
, , ,
, .
.-
2
(
2).
.-
3, 2
, NOS,
-
(Pryor and Squadrito, 1995).
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,
,
(NADPH ,
SOD, MPO iNOS).

,

ATP,
. , ,
.-
2 , 22
(Cadenas and Davies, 2000; Chance, Sies et al., 1979). ,
.
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,
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( 3).
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,
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22.
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2 (Chance, Sies et al., 1979).
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,
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2000). , ,
2 2.-.
,
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Q .
, , 2 22, Cyt-C
p66shc
(Giorgio, Trinei et al., 2007). , -,
,
, 22 (Cadenas, 1989; Hauptmann, Grimsby et al., 1996).
2.- 22
.

,
, (Cadenas and Davies, 2000).

,
, 2.- 22 NADPH
NADH. 2.- P-450.
,
P-450. ,
.
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b-5 , , .
b-5 2 2.-
.
, , ,
. ,
, 2.-,
, DNA.

,
, ,
.
, 5-6, 8-9, 11-12 14-15.

2 (Sevanian and Kim, 1985).

. ,
( 2),
, ,
( 4).
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= , = , LTA4 = , PG2 = .
PGG2 PGH2
, (PGHS). PGG2
PGH2 () ()
, ESR.
.
Ox (.),
.
. (
)
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, ,
.
, (Fe+4=O),
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. , ,
,
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3%
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:
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, , 2.-.
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, .
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,
.

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, ,
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, ,
.
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, (XDH),
.-
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, (-SH)
. K , in vivo ,
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, ,
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2 , , , ,

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, , (NADPH NADH)
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6: .
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, ( ). , ,
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, , ,
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. 6
, ,
. , , 2,
2.-. 2.-
(NADPH, NADH) 2, ,
. ,
,
.

, , ,
.
.
, 7.
7: . ,
.

.
, , , ( 7).
,
,
-450.
,

.

. , ,
,
(Galaris, Georgellis et al., 1985).
, ( ),
, ,
, .
,
, NADPH ,
Ca2+-ATP ,
. ,
,
( --p-) .
, , ,
, .
, ,
( , ,
...).
-
. , ,
.
, ,
,
,
.

, ,
( SH ), , ,
L-DOPA (), (FMNH2 FADH2) ,
2, , 2.-. 2.-, ,

. ,
,
.

(200-400 nm)
(-), ( 1),
(*), , ,
( 2-4):
(
-) (, , -).
, ,
( 5-7):
7 (*),
( 8)
2
, ,
, ( 9):
2.+ ,
(.), e- 2 (e-aq). H e-aq
.

. , (. e-aq)
, . , N2,
e-aq . ( 10):
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aging. Free Radic Biol Med 29(3-4) 222-230.
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Physiol Rev 59(3) 527-605.
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cultured heart cells from neonatal rats. Biochem Pharmacol 34(7) 989-995.
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oxidants, myeloperoxidase, and bacterial killing. Blood 92(9) 3007-3017.
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monoamine oxidase A/B causes oxidative damage to mitochondrial DNA. Arch Biochem Biophys
335(2) 295-304.
Kappus, H. and Sies, H. 1981. Toxic drug effects associated with oxygen metabolism: redox cycling
and lipid peroxidation. Experientia 37(12) 1233-1241.
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nitric oxide with superoxide. Am J Physiol 268(5 Pt 1) L699-722.
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membranes. J Free Radic Biol Med 1(4) 263-271.
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1

.
,
. ;
,
;
, NADPH
.-
2 2 ,
(respiratory burst).
(oxidative burst),
, .
O2 NADPH
NADPH gp91phox
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, H2O2 SOD .
, , 2.- 22 ,
. ,
,
(). 22
(HOCl):
,
, .
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(CGD, chronic granulomatous disease)
NADPH .
, , ,
, .
.-
2
(
2).
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, , 2
, NOS (iNOS).
,
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- -
, OCl, NO2 NO2Cl.
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.
;

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.
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, , ,
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(SOD) 1969
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.
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2), ,
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,
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, 2.- ,
. 2.-
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2.-.
, Haber Weiss 1932 ( 1):
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Cadenas et al., 1989; Galaris, Sevanian et al., 1990).
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), , ,
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DNA RNA, , ,
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DNA (Tanaka, Chock et al., 2007).
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,
,
. 100
(Dizdaroglu, 1991; Lindahl, 1993).
DNA .
: (i)
(direct repair), (ii) (base excision repair), (iii)
(nucleotide excision repair), (iv) (mismatch repair), (v)
(homologous recombination repair) (vi)
(non-homologous end-joining repair) (Sancar, Lindsey-Boltz et al., 2004).

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.
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.

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, ,
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,

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( 9) .
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.

(Boschi-Muller, Gand et al., 2008).
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.
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(Drazic and Winter, 2014).
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. , , ,
(Moskovitz, 2005; Stadtman, Van Remmen et al., 2005).
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, 22.
. ,
. .
, ,
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.

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, (Feeney and Schoneich, 2012).
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.
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.

., ROO. RO. , ,
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, - 2., .

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4--, ,
(LDL).
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, .
,
.
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, (Ehrenshaft, Deterding et al., 2015).
.

Ca2+ + (Ca2+-ATP, Ca2+/Na+-ATP Na+/K+-ATP).
(-SH)
.
, , ,
, (..
, 4-- ...) (Higdon, Diers et al., 2012).
.
, ,
.
, ,
, (L.) (LO.) .

.
.

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. , Na+/K+-ATP
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.

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.

.
,

.
.
Kelvin Davies
15 20S
(Davies and
Shringarpure, 2006).
,
. ,
22
.
Cu,Zn-SOD,
.
22 Cu .
.
22

.
20S ( 670 kDa)
( 26S, 1500 kDa)
.
,
.
Ahel, I., Rass, U., El-Khamisy, S.F., Katyal, S., Clements, P.M., McKinnon, P.J., Caldecott, K.W.
and West, S.C. 2006. The neurodegenerative disease protein aprataxin resolves abortive DNA
ligation intermediates. Nature 443(7112) 713-716.
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and substrate specificities. Arch Biochem Biophys 474(2) 266-273.
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discovery and study of sources and targets of hydrogen peroxide redox signaling through NADPH
oxidase proteins. Annu Rev Biochem 84 765-790.
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proteasome may be inhibited in aging and in inflammatory neuromuscular diseases. Neurology 66(2
Suppl 1) S93-96.
Dizdaroglu, M. 1991. Chemical determination of free radical-induced damage to DNA. Free Radic
Biol Med 10(3-4) 225-242.
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Biophys Acta 1844(8) 1367-1382.
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apoptosis in human monocytes. Biochem J 435(1) 207-216.
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tryptophan residues by reactive oxygen species, modes of detection, and biological consequences.
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potential protective mechanism against oxidative reperfusion injury in muscle. Arch Biochem
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1
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1969; McCord and Fridovich, 2014).
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, FeSOD .
SOD , Cu Zn,
.
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( 2).
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.
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.
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,
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= 2 x 109 M-1s-1).
(MnSODs)
Mn (Mn-SODs),
CuZn-SOD ,
. 40 kDa,
CuZn-SODs 32 kDa. SODs
, . .
,

, CuZn-SOD Mn-SOD, .

.
, , ,
ATP.
SODs
(Fe-SODs) , E. Coli,
. Mn-SODs
CuZn-SODs.
Mn-SOD/Fe-SOD.
. , , SODs ,
.
SODs.
Fe-SOD.

.
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, CuZn-SOD
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SOD,
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, SOD
.
McCord Fridovich 1969
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2 550 nm.
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..
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Mn-SODs,
.
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,
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. ,
. 4,
(22) . ,
22 2, O2.
4:
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(), 22 ().
,
(Fe5+) ( e-) (Fe3+).
.
( -Fe4+=O) ,
.
,
, 22
, , , . ,
Vmax . 22,
, , ,
, .
22
240 nm 2
. ( 5)
in vitro in vivo .
5: (3--1,2,4-).
, ,
H2O2 . ,
22
in vivo .

,
22.
22 .
(GPx)
, ,
22. GSH (GPx)
(GSH) 22 2
( 6).
6: 22 (GPx) ()
(GSH) (). H GSH a ,
. -
- ,
.
(GSH). ,
, , ,
22. , GPx , , ,
,. ,
( 3), R
.
4
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) ( -Se ) ,
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) GSH , GSSG,
( 6):
GSH
in situ .
, .
Fulvio Ursini
, -
GSH in situ
(Ursini, Maiorino et al., 1985). ,
GSH 22
,
(Brigelius-Flohe and Maiorino,
2013).
GSH , Se
. GSH.
,
.

GSH.
.
GSSG NADP+
,
(GSH/GSSG) 50 100.
, ,
, . ,
GSSG GSH.
, GSH (GR),
7 ( 7).
7:
. PGP = , pH =
pH .
NADPH, 7,
(PGP)
.
6-
(G6P-DH), 8. , 6-
(G6P) 6- ADP+ NADPH.
8 ,
6- ( 9) 5-
NADPH.
NADP+
6- (G6PDH). GSH
NADPH/NADP+ ,
NADPH.
NADPH NADH-NADPH-
,
10, (pH)
.
,
GSH, NADPH,
. , ,
6- ( ),
NADP+. , , ,
22 . 22
NADP+, GSH/GSSG
, GSH .
,
() ( )
. ,
,
. , ADPH
, GSH , ,
ADPH.
, , ,
.
(Prx)

(Prx)
(Chae, Oubrahim et al., 2012). 20-30 kDa
, GSH,
. ( 1%
) ,

(Rhee, Chae et al., 2005).
,
Saccharomyces cerevisiae
. ,
(Chae, Chung et al., 1994).
6 (PrxI
PrxVI). ,
: () 2Cys Prx (Prx1 Prx4), () 2Cys Prx (Prx5) ()
1Cys Prx (Prx6).
, 22,
, 2 , .
( 8).
8: . Sp SR
, .
(SP, pKa)
(SR) . ,
SP- . (
, k = 105-107 M-1s-1 pH=7.4).

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Glu/Gln/His). Sp
,
(Hall, Nelson et al., 2011). (SPOH), ,
SR
. (Trx),
(TrxR),
NADPH.
22 , (-
CH2-SPH) , (-CH2-SOH), (-CH2-
S2H) , , (-CH2-S3H). ( )
,
. , ,
, (Srx),
ATP (Rhee, Jeong et al., 2007).

(Prx), , ,
(redox signaling).
(Trx) (Grx) (Ahsan, Lekli et al., 2009; Lillig, Berndt
et al., 2008; Lillig and Holmgren, 2007; Rhee and Woo, 2011).
, .

. (TrxR, thioredoxin reductase),
(GR, glutathione reductase) (Prx, proxiredoxin)
,
( 9).
9: ,
, NADPH (Trx) /
(GSH/Grx). TrxR = , Prx = , GR = , Grx =
, PSSG = , PSH = , PSSP =
( ).
NADPH,
. ,
,
(TrxR), (Se) (S) . Se
, ,
.
(Trx) ~12 kDa
.
, , .

/ (PSSX).

22 2, . , ,
(
), ( 9).

(Cys-X-X-Cys) .

.
,
NADPH,
. ,
,
, .
,
,
DNA ( ),
, ASK1, AP
(Matsuzawa and Ichijo, 2008; Saitoh, Nishitoh et al., 1998).
, , NF-B AP1,
, 22 (Abate, Patel et al., 1990;
Matthews, Wakasugi et al., 1992).

,
GSH, GSH
(PDI), (Hanschmann, Godoy et al., 2013).
Cys-X-X-Cys ,
,
. , ,
. ,
.
,
. , ,
.

(TrxR) GSH.
, 50 60 kDa,
(subunits) - (FAD) NADPH.
,
(Se), , (Rundlof, Janard et al., 2004).
Se
S , NADPH FAD2.

, . ,
GSSG . ,
.
,

() ( 10).

ASK1 (apoptosis signal-regulating kinase 1). ASK1 MAPKKK,
, , ,
MAP JNK p38 (Ichijo, Nishida et al., 1997). ASK1
/ ,
ASK1 ( 10).
ASK1
(Song, Rhee et al., 2002).
,
, MAP JNK p38.
,
(Cys67) p50 NF- (
p65 p50).
( )
( 10). ,
NF- ()
, NF-
.
10:
.

, GSH,
22, ,
. , ,
.
22
;
GSH
. , .
, GSH
22, ,
22.

, . 22
,
,
,
. GSH
GSH , .
,
GSH. , , , GSH
(matrix).
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) ,
, ,
GSH. .
22 ,
. , ,
.
,
(redox signaling).
, 22.
1 (NADPH
) 22 PDGF TGF
.
. ,
( ) 1 .

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GSH ),
( ,
),
( GSH, DT-),
( 2, ),
(, ),
. , .

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GSH
, GSH.

,
, -450.
, ,
.

GS/ ,
,
ATP.

, , , , DDT, ,
.
, ,
.

. , ,
. ,
- .

GSH
, .
, GST.
,
.
, , ( )
. , , C4
GST, , , .

GSH, ,

. 22 2,
.
22 ( 11):
- ( 22 2),
,
(Fe4+=O),
. -
, . ( 12)
( 13):
2.
1 2
22 (Cl-, Br-)
22 (SCN-)
22 -
22
22 C
Horse radish peroxidase 22
2.
,
, ( 2).
( 4).
-
, 22 Cl ,
. (HOCl)
( 14):
(Cl-) ,
.

,
(SCN-) (OSCN-).
.
22, , . .
, ,
.
(-) ( ) .
22 NADPH ,
.

22 ( 15):
- ,
.
(Asc ). ,
,
( 16):
, ,
.
(disproportionation reaction).
, , ,
, , 22 2
, (Galaris, Cadenas et al., 1989;
Galaris, Eddy et al., 1989). , , ,
,
22,
(Galaris, Mira et al., 1988; Galaris, Sevanian et al., 1990).
NAD(P)H: 1 (NQO1, DT-)
NAD(P)H: 1 (NQO1, DT-)
(Siegel, Yan
et al., 2012). , ,
, , , NADH
NADPH.
,
.
.-
(2 ),
( 11).
11: NAD(P)H: 1 (NQO1, DT-)

.
, , ( ,
), 2.- 2.
, , ,
,
(Siemankowski, Morreale et al., 2000).

.
(, DNA, ) ( 5). ,
.
,
12.
12: .
, ,
, DNA.
, .
.
,
( DNA )
().

,
, . , ,

. ,
, 22,
,
.
.
, 22 oxyR.
oxyR OxyR 9
, ( 13). ,
OxyR . ,
22, OxyR
, DNA
.
oxyR , , GSH,
DNA (heat shock protein) DNAK.

. , oxoR,
E. Coli.
2.- 22, , SoxR
( 13). SoxR ,
, Mn-SOD, V, 6-
C.

.
.
Nrf2 (Forman, Davies et al., 2014),
.
Nrf2 ,
. 9 10.
13: 22 2.- . ()
OxyR 22 ,
DNA
, ..
. () SoxR DNA
SoxR. 2.- 4Fe4S
SoxR,
(.. SOD).

Abate, C., Patel, L., Rauscher, F.J., 3rd and Curran, T. 1990. Redox regulation of fos and jun DNA-
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paradigm for oxidative stress. IUBMB Life 48(1) 41-47.
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potential protective mechanism against oxidative reperfusion injury in muscle. Arch Biochem
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Galaris, D., Mira, D., Sevanian, A., Cadenas, E. and Hochstein, P. 1988. Co-oxidation of salicylate
and cholesterol during the oxidation of metmyoglobin by H2O2. Arch Biochem Biophys 262(1) 221-
231.
Galaris, D., Sevanian, A., Cadenas, E. and Hochstein, P. 1990. Ferrylmyoglobin-catalyzed linoleic
acid peroxidation. Arch Biochem Biophys 281(1) 163-169.
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glutathione peroxidase. Biochim Biophys Acta 839(1) 62-70.
1

.
;
,
(TrxR, thioredoxin reductase), (GR, glutathione reductase)
(Prx, proxiredoxin) ,
( 10).
NADPH,
. , ,
(TrxR), (Se) (S)
. Se ,
, .
(Trx) ~12 kDa
.
, , .

/ (PSSX).

22 2, . , ,
(
), .

(Cys-X-X-Cys) .

.
, , ,
NADH,
. ,
,
, .
2

;
,

() , ( 11).

ASK1 (apoptosis signal-regulating kinase 1). ASK1 MAPKKK,
, , ,
MAP JNK p38. ASK1
/ , ASK1
( 11).
ASK1
. ,
,
MAP JNK p38.
,
(Cys67) p50 NF- (
p65 p50), .

( )
( 11).
7 - :

. ,
(scavenger) . ,

, : ()
()
,
. ,
( ).
,
, ,

.
/ ,
6, ,
,
. (free radical
scavengers) ( 1).
.

( C)
- ( )
-

1.
,
,
( 1):
,
( , ) , ,
.
, , ( ). ( C) -
( )

, . , ,
, ,
. ,

. ,
,
, .
,
, . ,
Se, Mn, Zn, Cu Fe
, SOD, GSH
, , 2.- 22. ,

, GSH,
22 . ,
, ( Fe Cu)
,
,
1. .
()
(),
.
.

.
, ,

. , ,
( C), ( ),
.
( C)

,
.
,
,
.
,
, . ,

(Fe2+) , , . ,
,
. , ,
, Cu
.
Fe3+ Fe2+
,
, . , ,
(..,
Fe4+ ) , ,
(Galaris, Eddy et al., 1989; Galaris
and Korantzopoulos, 1997).

, 16 ,
, . ,
, , ,
, 1795
.
, ,
.

,
.
, .
()
, .
(GLUTI 1) .

Szent-Gyorgyi 1928. , ,
.
(p1 = 4.25 p2 = 11.8). , pH ( 1).
1: ( C) .

. , Fe Cu
, ,
.
. (Asc ) ,
( 1). , 2,
, ,

(disproportionation) ( 2):
.
( Asc = , Asc = DHAsc =
).

. pH
, Fe Cu.

. ,
,
. Fe Cu
,
, , DNA.

.
NADH , GSH
. , ,
, , . ,
GSH ,
in vivo
(Winkler, Orselli et al., 1994).
, 2.-,
(.) (Cl),
.
(-S ) (Galaris, Cadenas et al., 1989).
, , ,
. ,
(12), in vitro.
, ,
- ,

( 2).
2: - .
in vivo
, ,

in vivo.
,
, .
,
ex vivo , .
, . ,
, ,
.
,
.
,
(Gey,
Stahelin et al., 1987; Loria, Klag et al., 2000; Simon, Hudes et al., 2001). ,
,
,
.
,
, , ,
,
( ) ,
(Cook, Albert et al., 2007; Heart Protection Study
Collaborative, 2002; Hercberg, Galan et al., 2004; Lykkesfeldt and Poulsen, 2010).

. ,

.

Evans and Bishop 1922
(Evans and Bishop,
1922). , (
) , (,
, ) ( 3).
3: .
, .
,
. -, ,

E.
.

. -
(LDL)
(Esterbauer, Schmidt et al., 1997; Packer, Weber et al., 2001).
.
(LOO ) ( 3)
.
.
(-Toc = -, -Toc = -, LOOH =
.
, LOO = ).
.
- (-Toc )
( 4), ,
, (, 2).

,
TBA- . , ,
, ,
-.

. ,
.
.
,
.

, , VLDL LDL.

(Ricciarelli, Zingg et al., 2001). , ,
, 12 .
-,
, . ,

.
,
-, .
Angelo Azzi ,
, -
(Azzi, 2007; Fanali, Fasano et al., 2013; Zingg and Azzi, 2004; Zingg, Han et al., 2013).
-
,
-, in vitro in vivo
(Bowry, Ingold et al., 1992; Upston, Terentis et al., 1999).
-,
(Stocker, 1999).
-
.
. , , ( )
in vitro, .
-, ,
in vivo. -
in vivo
.

, (-) .

. .
, ROO OH
-
ONOO HOCl, in vitro (Rice-Evans, Miller et al.,
1996) ( 2, 4).

, , , ,
, , , , ,

, ,

, , ,
,

, , ,
, ,
p-

2.
4: .
,
,
, ,
(LIP, labile iron pool),
, , , C
,
. ,
. ,
.
,
. ,
. ,
( ) , ,
, .

.

, 1.0 .

( )
50 500 ,
(Gerogianni, Simos et al., 2015).
600
,
(Stahl, van den Berg et al., 2002). , 80-85%
8-12% .
. 1 , ,
,
.

( 5),
.
40 , .
40 ,
, , .
5: , .

. ,
. 50
, - .

(quench) (12).
- ,
.
In vitro
2 . , ,
.
.
(Car +) , ,
(disproportionation reaction) ( 5)
, , ( 6).
.
, (ROO ), ( 7)
(Krinsky, 1993).
,
in vivo .

,

.
(GSH) ,
. Glu, Cys Gly, Glu
, - -
( 6).
6: .

1.0 10.0 mM. (95%)
, , (-S-S-)
GSH (GSSG, GSH) (
). ,
, ,
GSH/GSSG.
GSH
, , , , ,
Ca2+ . ,
.
GSH/GSSG .

GSSG.
, , GSSG
, .

, ,
. , SH

( ) ,
( ), , Cu,
..
, .
,
, , ,
(GSSG), .
,
.
(GS ).
(GSSG), 7.
.
7: (GS ) () ().
2, GSO. (
), 2.- 22 (Galaris, Cadenas et al., 1989). ,
,
. , , ,
( 8):
, ,
in vivo. ,
-
(ONOO ),
(GSNO), GSH NO. (
-
) , , ONOO NO.

, , (GSH) (GSSG),
(GSSR) SH .
GSSG SH
( 9) S-:
, , -SH
,
GSSG in vitro.
, .
, GSSG
,
.

( 8) ,
-, -.
, () () ,
,
. . . -
, ROO , RO , OH, ONOO , HCl .. ,
,
Fenton. , ,
,
( ) . ,
,
( )
, .
8: .
( Q, CoQ)
Q (CoQ)
.
.
, , ,
-. ,
.
(ROO ),
.
Q - (
10), in vivo .
.
( -Toc -Toc = - -, CoQH2
.
CoQH = Q Q, ).
,
. , , CoQ
2 ( b C
.-
) , , 2 . ,
CoQ
in vivo.

Bruce Ames
, . ,
,
.

( 9):
9:
. ,
2 NAD+ 2.-
NADH.

. , , (,
), ,
(0.2-0.4 mM). ,
.
, ,
,
( ).
, .
. .
, RO , OH, 12 ,
.
(Ur ),
. 1981 Bruce Ames Paul Hochstein ,
,
in vivo. ,
. ,
, (3),
-
( ) (2). ,
Fe ,
Fenton. in vivo , ,
,
( )
(.., Wilson, , ..).
, ,
, ,
.
-1.
, , ,
.
( 10)
in vitro ,
.
(ROO ) (12).
10: .

1:1. /
, , .
,
, . ,
( 12)
.

Azzi, A. 2007. Molecular mechanism of alpha-tocopherol action. Free Radic Biol Med 43(1) 16-21.
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secondary prevention of cardiovascular events in women: results from the Women's Antioxidant
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1
( C)
;
,
,
in vivo.

, , .
,
ex vivo , .
, . ,
, ,
.

, .

,
(Gey, Stahelin et al., 1987; Loria, Klag et al., 2000;
Simon, Hudes et al., 2001). ,
,
,
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.

.
2
in vitro -
, .
-
in vivo;
,
- , in vitro in
vivo.
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.
. , , ( )
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.
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.
,
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.
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, .
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1930, .
1958 (Josephs,
1958). (TfR)
(IRP/IRE system) 1970 1980, .
,

.
(hepcidin), ,
(ferroportin), (Abboud
and Haile, 2000; Donovan, Brownlie et al., 2000; Krause, Neitz et al., 2000).
,
-
. ,
.

,
, , ,
, , DNA,
. , ,
.
,
, ,
. ,
,
,
(Weinberg, 2010).
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1), , ,
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.
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al., 2012). , ,
, C,
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, ,
.

(Hower, Mendes et al., 2009; Pietrangelo, 2007).
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, .
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. ,
,
.
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)
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( 1).

. ,
.
1: DNA
22. (Jurkat, -)
1,7- 1,10- (), 2,2- () SIH ()
22. DNA comet assay (Barbouti, Doulias et
al., 2001). DNA,
.
, ,
, Fenton. ,
Fenton ,
.
.

,
.

, ,
.

. (hepcidin),
(Ganz and
Nemeth, 2011).
( ~79 kDa)
1.2 2.0 mg/ml.
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2012). , , ,
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.
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.
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,
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.

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.
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-1
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( 8). pH
, Fe2+ Steap3
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Campagna et al., 2005).

.
- . ,
.
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TfR1 .

DMT1. ,
.
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(labile iron)
,
(~ 2-5%)
.
, , (LIP, labile iron pool),
, ,
..
, . , ,
, ,
.
,
.
, ,

,
. .
LO . , , ,

.
,
(IRPs, iron-regulatory proteins).
,
, (Wilkinson and Pantopoulos,
2014). IRPs (IRP1 IRP2)
,
mRNA
. mRNAs
(IREs, iron-responsive elements) ( 9).
mRNAs -1 (TfR1), ,
, mRNAs
. IRPs IREs
, IRE 5 3
mRNA. ,
, mRNA
. 9,
IRP1 3 mRNA
1, mRNA
. , IRP1 3 mRNAs
, .
. ,
, IPRs IREs,
.
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, Krebs
. [4Fe4S]
. IRP1, 4Fe4S ,
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,
,
, , ,
Fenton (.) (RO.).
,
, .
, IRPs
.
9:
, IRP/IRE. () , IRPs
(iron regulatory proteins)
IREs (iron responsive elements). (B) , IRPs
IREs. mRNA
(TfrR1), mRNAs
.

,
, (, ...).
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o ).
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).
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, , , .

(), () DNA. , ,
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.

, .

.

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.
.

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.
( 1-2 mg ),
,
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2
(Zimmermann and Hurrell, 2007).
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.

,
.

.
1
.

HFE HFE

HFE2
HAMP

TfR2 TFR2

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1.
, ,
,
.
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.
, : (i) , (ii)
, (iii) (NTBI) (iv)
.

. ,
(, ) ( ,
) (Sebastiani and Pantopoulos, 2011). ,
-
( SOD ),
.
,
,
.
,
( 1).
() HFE : (
1) HFE (Beutler, 2006; Pietrangelo,
2004), MHC 1 (Major Histocompatibility Complex, class 1).
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. HFE
(C282Y).
HFEC282Y ( )
. , HFE, H63D S65C,
, C282Y.
, ,
.
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, (Olsson, Lind et al., 2008).
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.
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.
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, .
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, .
,
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4
.
: ,
, (
4),
( 4).
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.
.

,
. ,
, ,
.

. ,
.
,
. ,
,
( ).

. , (,
) , ,
().
,
, , .

,
(Weiss and Goodnough, 2005)[Weiss and Goodnough, 2005].

. ,
.
,
.
,
-,
.
( 10).
10:

.
(IRIDA)
(IRIDA, iron refractory iron deficiency anemia)
TMPRSS6,
2 (Finberg, Heeney et al., 2008).
,

.
2 (
) 2 (Silvestri, Pagani et al., 2008).

.
, : (i)
, (ii) , (iii)
- (iv)
. (, 2010).
,
() .
, (NTBI)
,
. .

(< 1% ).
,

.
,
(Bartnikas, Andrews et al., 2011).
,
, .
( ),

.

, ,
(labile iron) ,
,
. ,
(.., ), (.., ),
. ,
,
.
(DFO), (DFP) ( 11).

, (
). ,

. , ,
.
11:
.
(DFO) 40
. Streptomyces
pilosus , ,
. ( )
, , ,
.
(Doulias, Christoforidis et al., 2003), DFO-Fe .

, (L1) .
, ,
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, . 2010.
. . , . 155.
1

: ,

,
.
;

(loss-of-function) (gain-of-function). ,
,
.
,
, ,
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2
Cu Fenton 60 , Fe.
( Fe)
;
, ( ),
. ,
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, .
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22 .
,

1950 (Gerschman, Gilbert et al., 1954; Harman, 1956). ,

. 1969, McCord Fridovich
,
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, ( 22)
. 22
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2
1990. , 22
,
(Finkel, 1998).

(EPR, electron paramagnetic resonance),
.-
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derived growth factor) EGF (epidermal growth factor), . ,
22, ( 22
2) ,
.
2: 22
.
22
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22
22 , ,
(Breton-Romero and
Lamas, 2014; Stone and Yang, 2006).
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enzymes) . 22 ,
, , ,
, P-450.

.-
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SODs. ,
, D- , 2
22 .
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,
, NOX
(Burgoyne,
Mongue-Din et al., 2012).
,
NADPH ,
.-
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. DUOX1 DUOX2, C-
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, (Cadenas and
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, 2,
.-
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.-
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p66shc
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2010). 1 2% 2
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2 (Chance, Sies et al., 1979). 2
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,
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,
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22 ,
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,
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,
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, . ,
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,
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.
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ASK1, apoptosis signaling regulating kinase) -
, (Trx). -
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like factor). , Nrf2,
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. Keap1
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Keap1 ,
.
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. , ,
.
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-
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().
, PG1 (Peroxy Green 1) PC1 (Peroxy Crimpson 1) ( 10).
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,
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-
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() (HOCl). (PG1
PC1) , 22
.
, (confocal microscopy),
22
(EGF) (Miller, Tulyathan et al., 2007).
,
22
, . ,
22 ,
22 .

, GFP (Green Fluorescence
Protein), 22. , roGFP1
roGFP2 GFP
22 (Rhee, Chang et al., 2010).
,
. ,
, ,
.
22 , GFP
22. ,
22 ( k = 105-107 -1s-1)
GFP. ,
( 22) .
GFR YFR (Yellow Fluorescence Protein).
22
22
.

22
.
(bioenergetics) 1920, Otto Warburg :
, 22. , Keilin
Hartree 1945 (Keilin and Hartree, 1945) : 22
, .
, Britton Chance 1951:
22 , 22,
, . 1970
22 (Chance, Sies et al., 1979; McCord and
Fridovich, 1969).

.
.
Bedard, K. and Krause, K.H. 2007. The NOX family of ROS-generating NADPH oxidases:
physiology and pathophysiology. Physiol Rev 87(1) 245-313.
Brand, M.D. 2010. The sites and topology of mitochondrial superoxide production. Exp Gerontol
45(7-8) 466-472.
Breton-Romero, R. and Lamas, S. 2014. Hydrogen peroxide signaling in vascular endothelial cells.
Redox Biol 2 529-534.
Burgoyne, J.R., Mongue-Din, H., Eaton, P. and Shah, A.M. 2012. Redox signaling in cardiac
physiology and pathology. Circ Res 111(8) 1091-1106.
Physiol Rev 59(3) 527-605.
Finkel, T. 1998. Oxygen radicals and signaling. Curr Opin Cell Biol 10(2) 248-253.
Finkel, T. 2011. Signal transduction by reactive oxygen species. J Cell Biol 194(1) 7-15.
Forman, H.J., Ursini, F. and Maiorino, M. 2014. An overview of mechanisms of redox signaling. J
Mol Cell Cardiol 73 2-9.
Harman, D. 1956. Aging: a theory based on free radical and radiation chemistry. J Gerontol 11(3)
298-300.
Hur, W. and Gray, N.S. 2011. Small molecule modulators of antioxidant response pathway. Curr
Opin Chem Biol 15(1) 162-173.
Keilin, D. and Hartree, E.F. 1945. Properties of catalase. Catalysis of coupled oxidation of alcohols.
Biochem J 39(4) 293-301.
McCord, J.M. and Fridovich, I. 1969. The utility of superoxide dismutase in studying free radical
reactions. I. Radicals generated by the interaction of sulfite, dimethyl sulfoxide, and oxygen. J Biol
Chem 244(22) 6056-6063.
Miller, E.W., Tulyathan, O., Isacoff, E.Y. and Chang, C.J. 2007. Molecular imaging of hydrogen
peroxide produced for cell signaling. Nat Chem Biol 3(5) 263-267.
Paulsen, C.E. and Carroll, K.S. 2010. Orchestrating redox signaling networks through regulatory
cysteine switches. ACS Chem Biol 5(1) 47-62.
Ray, P.D., Huang, B.W. and Tsuji, Y. 2012. Reactive oxygen species (ROS) homeostasis and redox
regulation in cellular signaling. Cell Signal 24(5) 981-990.
Reth, M. 2002. Hydrogen peroxide as second messenger in lymphocyte activation. Nat Immunol
3(12) 1129-1134.
Rhee, S.G. 2006. Cell signaling. H2O2, a necessary evil for cell signaling. Science 312(5782) 1882-
1883.
Rhee, S.G., Chang, T.S., Bae, Y.S., Lee, S.R. and Kang, S.W. 2003. Cellular regulation by hydrogen
peroxide. J Am Soc Nephrol 14(8 Suppl 3) S211-215.
Rhee, S.G., Chang, T.S., Jeong, W. and Kang, D. 2010. Methods for detection and measurement of
hydrogen peroxide inside and outside of cells. Mol Cells 29(6) 539-549.
Schieber, M. and Chandel, N.S. 2014. ROS function in redox signaling and oxidative stress. Curr
Biol 24(10) R453-462.
Sies, H. 2014. Role of metabolic H2O2 generation: redox signaling and oxidative stress. J Biol Chem
289(13) 8735-8741.
Stone, J.R. and Yang, S. 2006. Hydrogen peroxide: a signaling messenger. Antioxid Redox Signal
8(3-4) 243-270.
Veal, E.A., Day, A.M. and Morgan, B.A. 2007. Hydrogen peroxide sensing and signaling. Mol Cell
26(1) 1-14.
Winterbourn, C.C. 2013. The biological chemistry of hydrogen peroxide. Methods Enzymol 528 3-
25.
1
22, ,

; .
1990, , (EPR, electron

.-
paramagnetic resonance), (2 )
( PDGF EGF)
. , 22,
( 22 2) ,
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22 .
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/.
(tyrosine receptors) -
,

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.-
. 22, , 2 ,

. 100-1,000
, . ,
.
2
22 ;
;
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. 22
,
-
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(-CH2-SOH),
.
, ,
( ).
,
(GSH),
(1 10 mM). ,

( ). ,
.
, 22,
(-SO2H) (CH2-SO3H). ,

, . ,
.
10 -
20 , ,

. ,
.
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( ) . ,
. ,
, ,

.
( )
. , , ,
: () -
, () , , ()
,
, ()
, Keap1
Keap1/Nrf2.

.

. ,
(cAMP, Ca2+ ...),
, .

.
(460-377 ..) ,

. , 20
, .
20 ,
,
, , ,
(Aravanis, Corcondilas et al., 1970; Gerber, 1994).
, ,
(Knoops, de Groot et al., 2004; Linos, Holmes et al., 2007;
Trichopoulou, Costacou et al., 2003; Willett, 2006).
,
.
,

(Visioli, Bogani et al., 2005).
(, ,
) ,

(DeWeerdt, 2011; Dilis and Trichopoulou, 2010; Simopoulos, 2005).
,
,

. ,

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, .
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) ,
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,
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Halliwell and Gutteridge, 2007) . ,
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.

;

.
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,
. ,
, ,
.
, in vitro ,
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.
.
,
. ,
.

:
v , k ,
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.
, . , ( OH)
(22).

, k 109 M-1s-1 ( 1) (Haag and Yao,
1992).

. ,
.
, ,

(, ).
- (RO.),
,
k 4 20x107 M-1s-1 ( 2) (Buettner, 1993).
,
, , k. ,
,
( ),
.
.
,
in vivo .
(
), ,
(
22 2 ) (Forman,
Davies et al., 2014; Ursini, Maiorino et al., 1995). , ,
,
.

,

(Finkel and Holbrook, 2000).

,
,
,

(labile iron) (Barbouti, Briasoulis et al., 2010; Barbouti,
Doulias et al., 2001; Doulias, Nousis et al., 2005; Kitsati, Fokas et al., 2012; Melidou, Riganakos et al., 2005;
Nousis, Doulias et al., 2005).
( )
(DNA, , ...)
.
,
, , , ,
, .

.

( ). ,
( 1). ,
,
. .
, RO .
1: () . 1, 2 3
. ()
-, ,
.
, , (1 2
) ,
. ,
. ,

. ,
- ,
( 2).

. , ,
.
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, 2, ,
, . ,
, (-) ().
2: --
. ,

2.
, ,
.
,
Nrf2,

. Nrf2
Keap1,
Nrf2 ( 3).
3: Keap1/Nrf2
. (-)
Keap1 ( Michael). Keap1
Nrf2, ,
DNA, .
Keap1 Nrf2,
. Nrf2
DNA (AREs, antioxidant response elements, EpREs, electrophile response
elements) , , ( )
. ,
( )
( ), ,
DNA (Forman, Davies et al., 2014).
,
,
. , .. -
-, (
). : ()
,
, ()

Nrf2.

IRP1 IRP2 (Iron Regulating Protein 1 2),
,
,
(Galaris and Pantopoulos, 2008; Pantopoulos, 2004).
, , , ...,
, ,
,
.
, ,
.
,
, .
,
.

, .
(
) .
,

. ,
( ) ,
(
) . , ,
.

Keap1/Nrf2

, ,
, .

Nrf2 (nuclear factor E2-related factor 2).
,
Keap1, . Keap1 27 ,
.
, Nrf2.
Keap1 Nrf2
, DNA, AREs (antioxidant
responsive elements) , , EpREs (electrophile responsive elements),

(Dickinson, Iles et al., 2003; Moinova and Mulcahy, 1998; Mulcahy and Gipp, 1995).
Nrf2 , ,
(SOD), (CAT), 1 (HO1), ,
(GSH-S-transferase), NAD(P)H: (NQO1),
.

,
Nrf2
(Hur and Gray, 2011). Nrf2
Keap1 Nrf2 (Dinkova-
Kostova, Holtzclaw et al., 2005). , ,
,
Nrf2. ,
, ,
,
,
.

, .
, ,
- . :
() , () ()
().
()
.
, .

(~100 ),
. , ,
. ,
( 4).
4:
.
, .

.
,
, , ,
(Kitsati, Fokas et al., 2012).
()
(Kitsati, Fokas et al., 2012).
.
, pH
,
.
() ,
,
. ,
- .

5,
: ()
, ()
(),
,
. ,
Keap1/Nrf2
.
5: .
, : ()
-, () , ,
, () ,
, ()
, Keap1 Nrf2.
ICPs, (Iron Chelating Phenols).

Aravanis, C., Corcondilas, A., Dontas, A.S., Lekos, D. and Keys, A. 1970. Coronary heart disease in
seven countries. IX. The Greek islands of Crete and Corfu. Circulation 41(4 Suppl) I88-100.
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Preedy and R. R. Watson eds. Olives and Olive Oil in Health and Disease Prevention. Oxford,
Academic Press. pp. 1103-1109.
490-498.
Buettner, G.R. 1993. The pecking order of free radicals and antioxidants: lipid peroxidation, alpha-
tocopherol, and ascorbate. Arch Biochem Biophys 300(2) 535-543.
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AP-1 binding complexes and elevates glutamate-cysteine ligase gene expression. FASEB J 17(3)
473-475.
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140(7) 1274-1279.
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electrophiles and oxidants that regulates the phase 2 response, is a zinc metalloprotein. Biochemistry
44(18) 6889-6899.
Doulias, P.T., Nousis, L., Zhu, B.Z., Frei, B. and Galaris, D. 2005. Protection by tropolones against
H2O2-induced DNA damage and apoptosis in cultured Jurkat cells. Free Radic Res 39(2) 125-135.
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408(6809) 239-247.
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drinking water contaminants. Environmental Science & Technology 26(5) 1005-1013.
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Oxford University Press.
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Mediterranean diet and survival in a Greek population. N Engl J Med 348(26) 2599-2608.
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110.

1
, ,

( ). ; .
v k ,
[] [] ( )
, .
, (.OH)
( 22).

, k 109 M-1s-1 ( 1):

. ,
.
,
, , (k). ,
,
( ),
.
.
in vivo
.
( ),
,
( 22 2
). , ,
.
2

,
;

, :
() -
,
() , ,
() ,
,
() ,
Keap1 Keap1/Nrf2.
,
.

1
Fenton
(free radical)
(free radical theory of aging)
(oxidative stress)
(spin)
2
2 (singlet 2)
(second messenger)
3
(endothelial derived relaxing factor)
(NO-synthase)
(NO synthases)
4
(respiratory burst)
(oxidative burst)
(chronic granulomatous disease)
5
(direct repair)
(sister chromatid exchange)
(chaperones)
(in situ)
(mismatch repair)
(base excision repair)
(nucleotide excision repair)
(non-homologous end-joining repair)
(homologous recombination repair)
(redox signaling)
(signal transduction)
. (. scavengers)
(conjugated dienes)
6
(disproportionation reaction)
(glutathione reductase)
(thioredoxin reductase)
(redox signaling)
(nitroblue tetrazolium)
(peroxyredoxines)
(heat shock protein)
7
(quench)
(disproportionation)
(free radical scavengers)
8
MHC 1 (major histocompatibility complex, class 1)
(hepcidin)
(haephestin)
(IRIDA, iron refractory iron deficiency anemia)
(LIP, labile iron pool)
(heme oxygenase)
(iron-regulatory proteins)
- (chaperones)
(iron regulating proteins)
(non transferrin bound iron)
(non transferrin bound iron)
(iron responsive elements)
(ferroportin)
(bilirubin)
(biliverdin)
9
(labile iron)
10
(labile iron) 6

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- -

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10: PG1 PC1 22.

1990, , (EPR, electron

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