Institute of Experimental Pharmacology,

Slovak Academy of Sciences, Bratislava, SK

Pathophysiology and Pharmacology

of Reactive Oxygen Species (ROS)
Adaptation by
Wiwi J

Lab of Anaesthesiology & Intensive Treatment FK Unibraw/

RS Saiful Anwar - Malang

Reactive Oxygen Species as Mediators

of Tissue Injury, Diseases
and their Pharmacology
1

The Beginnings

1775 Joseph Priestley:

- discovery of O2

- observation of toxic effect of O2

1900 - Gomberg:
discovery of triphenylmethyl radical

Until 1950 60 :

minimal attention was given to biological

actions of free radicals and reactive
oxygen species
2

Evidence on the existence of ROS

1954 - Gerschman et al. : Recognition of

similarities between radiation and oxygen toxicity

1969 - McKord and Fridovich: Discovery of
superoxide dismutase and suggestion of the existence
of endogenous superoxide
1973 - Babior et al.: Recognition of the
relationship between superoxide production and
bactericidal activity of neutrophils
1981 - Granger et al.: recognition of the
relationship between local ROS production and
ischemia/reperfusion induced gut injury
3

 Free radicals have one or more unpaired electrons

in their outer orbital, indicated in formulas as [].

As a consequence they have an increased reactivity
with other molecules. This reactivity is determined
by the ease with which a species can accept or
donate electrons.
The prevalence of oxygen in biological systems
means that oxygen centered radicals are the most
common type found.
O2 acts in a process that is central to metabolism in
aerobic life, as a terminal electron acceptor, being
reduced to water. Transfer of electron to oxygen
yields the reactive intermediates.
4

 The term reactive oxygen species (ROS) rather

than oxygen radicals is now generally preferred

because singlet oxygen (its one form), hydrogen
peroxide,
hypochlorous
acid,
peroxide,
hydroperoxide and epoxide metabolites of
endogenous lipids and xenobiotics have chemically
reactive oxygen containing functional groups, but
are not radicals and do not necessarily interact
with biological tissues via radical reactions.
Molecular oxygen is a biradical, having two
unpaired electrons of parallel spin. As it is a
terminal electron acceptor being reduced to water,
oxygen acts in processes that are central to
metabolism in aerobic life.
5

4O2

XO

4O2
4e-

4e-

at presence
of NADPHO

Cytochrome Cascade

4O2-

4O2.-

4e4O224e-

SOD

4O234e2H2O2 + 2O2

CAT

cytochrome c-oxidase

Reactive Oxygen Metabolite

Cascade (ROM Cycle)

4O2416H+

2H2O +

4O2 + 4H + 4e
+

O2

2H2O + 3O2

8H O

4O2 + 16H2 + + 16e8H2O

(yields to production of ATP)

Half-life of some reactive species

Reactive species

Half-life
(s)

Physiol conc.
(mol/l)

Hydroxyl radical (OH)

10-9
Alcoxyl radical (RO)
10-6
Singlet oxygen (1O2)
10-5
0.05 1.0
Peroxynitrite anion (ONOO-)
7
Peroxyl radical (ROO)
1 - 10
Nitric oxide (NO)
minutes/hours
Semiquinone radical
spontan. hours/days
Hydrogen peroxide (H2O2)
(accelerated by enzymes)

Superoxide anion (O2-)

Hypochlorous acid (HOCl)

spontan. hours/days

10-9
10-9 - 10-7
10-12 - 10-11

(by SOD accel. to 10-6)

dep. on substrate

 ROS

present in mammalian tissues have both

endogenous and exogenous origin. Their
production is essential to normal function or
metabolism of most mammalian cells.
Approximately, 90% of all oxygen consumed by
mammalian cells is catalytically reduced by four
electrons to yield two molecules of water. It is now
clear that oxygen may also be reduced by less than
four electrons in enzymatic and nonenzymatic
reactions.
ROS are, however, also destructive unless tightly
controlled. Mammalian cells have developed a
battery of defenses to prevent and repair the
injuries caused by oxidative stress.
8

Origin of ROS
Generation in mammalian organism
Sources
endogenous
exogenous
prostaglandin synth.
respiratory chain
autooxidation

radiation, ultrasound
cigarette smoke
drugs

.NO

O2

e-

OONO-

O2.-

e-

heat
pesticides
infections
hyperoxia, exercise
air pollution (NOx, O3)

e-

.OH

H2O2

HOONO

NO2.

e-

H2O

O2.- O2 Fe2+ Fe3+ H+

FREE RADICAL S
phagocytes
oxyhemoglobin
oxidative enzymes
accumul reduced.metab.

H+

Myeloperoxidase

ClH2O

HOCl

O2 + Cl-

H 2 O 2 H2 O

Enzymatic sources of ROS

Xanthine oxidase
Hypoxanthine + 2O2 Xanthine + O2.- + H2O2
NADPH oxidase
NADPH + O2 NADP+ + O2.Amine oxidases
R-CH2-NH2 + H2O + O2 R-CHO + NH3 + H2O2
Myeloperoxidase
Hypohalous acid formation
H2O2 + X- + H+ HOX + H2O
NADH oxidase reaction
Hb(Mb)-Fe3+ + ROOH Compound I + ROH
Compound I + NADPH NAD+ Compound II
Compound II + NADH NAD+ E-Fe3+
NAD+ O2 NAD+ + O2.Aldehyde oxidase
2R-CHO + 2O2 2R-COOH + O2.Dihydroorotate dehydrogenase
Dihydroorotate + NAD+ O2 NADH + O2.- + Orotic acid

10

Effect of superoxide dismutase (SOD) on reduction of

cytochrome c by O2.- which is produced by xanthine
oxidase (XO) in the presence of xanthine (X)
no SOD

Absorbance

550 nm

0.06
0.04

XO 2 mU/ml
0.02
0.00

+ SOD 200 U/ml

-0.02

X100 mol/l
0

100

200

300

400

500

600

700

800

900

time (s)
Institute of Experimental Pharmacology, SASc, Bratislava, SK

11

Nonenzymatic sources of ROS

and autooxidation reactions
Fe2+ + O2 Fe3++ O2.Hb(Mb)-Fe2+ + O2 Hb(Mb)-Fe3++ O2.Catecholamines + O2 Melanin + O2.Reduced flavin
Leukoflavin + O2 Flavin semiquinone + O2.Coenzyme
Q-hydroquinone + O2 Coenzyme Q (ubiquinone) + O2 .Tetrahydropterin + 2 O2 Dihydropterin + 2 O2.12

 Until the 1960s, free radicals were not considered

particularly relevant for mammalian physiology

and pathology.
The discoveries of the existence of superoxide
dismutase (SOD) activity in mammalian cells in 1969
by McCord and Fridovich and association of
bactericidal activity of neutrophils with production
of the superoxide radical (O2.-) by Babior and
coworkers in 1973, linked free radicals to numerous
physiological and pathophysiological processes.
One decade later, in 1981, Granger and coworkers
established a hypothesis on the role of these reactive
species in the reperfusion injury after intestinal
ischemia.
13

ROS are tightly controlled

resulting in a physiological balance between their

production and elimination

Enzymes: SOD (c, m) ,

GPX (c, m), CAT (c, p)
Non-enzyme antioxidants:
vitamines (E,A,C), thiols,
phenols, ceruloplasmin,
transferrin, uric acid,
albumin, etc.

ROS:
O2, H2O2 , 1O2

OH, HOCl

c-cytosolic, m-mitochondrial, p-peroxisomal

14

Biological antioxidant defense mechanisms

b. Free radical scavengers (antioxidants)

Vitamin E (-tocopherol) O2.-, .OH, LPO
Reduced ascorbic acid

in high concentrations of O2.-, .OH, LPO

Low m.w. thiols (e.g. cystein)
Large m.w. thiols (e.g. albumin)

LPO

)
(Cu/Zn-Mn

E
Fe

H2O2

c. Removal of Fe and Cu
Ferritin, transferrin, lactoferrin (Fe)
Ceruloplasmin (Cu, Fe)
Serum albumin (Cu)
H2O + O2

2+

Vit
am
in

utases

- superoxide dismutase (SOD)

- ceruloplasmin
H2O2 - glutathion peroxidase (GTPx)
- catalase (CAT)
Organic hydroperoxides - GTPx
Disulphide - GTPx
Oxidised ascorbate - GTPx

O2.dism
Superoxide

a. Catalytic free radical removal

O2.- - spontaneous dismutation

Catal
ase

Defense mechanisms in the organism

Fe3+

OH

Ferritin

GS
HPe
rox
GSH
ida
GSSG

se

H2O

15

Under pathological condition

the physiological balance is lost

Enzymes: SOD,GPX,CAT
Non-enzyme antioxidants:
vitamines (E,A,C), thiols,
uric acid, ceruloplasmin,
transferrin, phenols,
albumin, etc.

ROS:
O2, H2O2 , 1O2,

OH, HOCl

16
Consequences are shown in the next panels

 Disbalance between production and elimination of

ROS
develops
during
inflammation,
ischemia/reperfusion, altered metabolism, action
of drugs, pollutants, etc.
Such disbalance causes pathology of brain, heart,
vessels, gut, airways, muscle, parenchy- matous
organs (liver, kidney, pancreas), eye, skin, joints,
etc.
Exposure of the tissues to ROS in a variety of
biological systems has documented their ability to
damage lipids, proteins and DNA. The resulting
damage potentiated by increased free intracellular
Ca2+ causes activation/deacti-vation of various
enzyme systems and cell injury or death.
17

Mechanisms of ROS induced cell injury

LIPIDS

Lipid
peroxidation

DNA damage
PROTEINS

Oxidation of thiols
Carbonyl formation

Damage to Ca2+ and

other ion transport
systems
Membrane
damage

DNA

Schiff bases

SUGARS

Poly ADP Altered gene

ribosylation expression

Instability to maintain
normal ion gradients

Amadori products
Depletion of ATP
and NAD(P)(H)

Activation/Deactivation of
various enzyme systems

AGEs

(Advanced glycation
end products)

Cell injury

18

Involvement of ROS in APOPTOSIS

NOXA
(trauma, hypoxia
metabolic insufficiency
activation of excitatory receptors)

under homeostatic
control to a certain
limit

Ion disbalance
ROS generation

Mitochondrial failure

caspase/calpaine
activation

Bcl-2 / Bax disbalance

CELL DEATH (necrosis / apoptosis)
19

 Currently it is believed that free radicals are

definitely participating in several health

disorders.

There are different pathologic conditions

where extracellular, intracellular or both

ROS act at least in part.

However, in spite of the extensive studies our

knowledge concerning the role and action of

free radicals and ROS is still incomplete and
changing.

20

Pathological conditions that may have a free radical

component and sites of ROS actions
Hypo-, hyperoxygenation
&
Immune ReperfusionCataractogenesis
after
reactions
ischemia
Chemical
Radiation injury
cancerogenesis
Ageing &
Cancer
senile dementia
Diabetes
Atherosclerosis
Parkinsonism
Iron, drug
& chemical
Smoking,
toxicity
air pollutants
Inflammatory
&
reactions drug induced
reactions

FREE
RADICALS

intracellular

Intracellular &
extracellular

extracellular
21

ROS generation during ischemia and reperfusion

ATP
I
S
AMP
Xanthine dehydrogenase
C
H
Adenosine
E
Ca2+ proteases
M
Inosine
I
A Hypoxanthine+Xanthine oxidase O2.- activated chemoattractants
REOXYGENATION
Cl-

H2O2

MPO
HOCl

O2.-

Fe2+
.

OH

Tissue damage

Extravasated
neutrophils

Circulating
neutrophils

Fe3+
Neutrophil
activators

Chemoattractants
22

ROS in the sequence of events in

STROKE
HYPOXIA
ATP depletion
Cell depolarization
( Mg block of NMDA rec.)
Excitatory aminoacid release
Ca2+ influx into the cells
Activation of phosholipases,
proteinkinases, proteases,
endonucl., phosphatases etc.
ROS generation
ONOO- generation
Devastatory effect in cells

Slow accumul. Ca2+ in mitochondria

MPT pore opening in mitochondria
H+ gradient collapse in mitochondria

NEURONAL DEATH
Therapeutic interventions: cyklosporine (specific MPT pore inhibitor),
antioxidants (lazaroids, deferoxamine, SOD in liposomes, allopurinol) 23

Frequent targets of ROS

airways

gut

O2-

OH

HOCl

H2O2

brain &
nerves

heart &
vessels
24

 ROS affect different tissues and tissue

components.
They affect e.g. not only the smooth
muscle cells, but also their epithelium,
endothelium, innervation, membrane
lipids, receptors, transmitter systems,
prostanoid production, Ca2+ homeostasis, etc.
25

longitudinal muscle

untreated

10 mN

5 min

whole ileum

whole
ileum

Effects of H2O2 on guinea pig

0.5 mmol/l H2O2
ileum

1mol/l Hi

atropine & guanethidine

26
treated
Institute of Experimental Pharmacology, SASc, Bratislava, SK

Effects of various ROS on guinea pig trachea

H2O2

OH (H2O2 + FeSO4)

1 mN
2 min

OH (FeSO4 + Ascorbic acid)

O2 - (Xanthine + Xanthine oxidase)
27

Institute of Experimental Pharmacology, SASc, Bratislava, SK

Proposed mechanisms of ROS actions in airways

In physiological conditions
O2.-

O2.-

No changes

H2O2

Dominant
contraction

OH

Diameter
of trachea

Dominant
relaxation

Resting
tone

Initial
Late
change
change
of tone
of tone
after ROS after ROS

SOD- superoxide dismutase; Cat catalase;

LMWAO low molecular weight antioxidants

epithelium
smooth muscle

OH

Diameter
of trachea

H2O2

In pathological conditions

Resting
tone

Dominant
contraction

Long-lasting
contraction

Intensive
contraction

Initial
Late
change
change
of tone
of tone
after ROS after ROS

28
Institute of Experimental Pharmacology, SASc, Bratislava, SK

 NO reacting with O2- gives rise to

unstable peroxynitrite, which decomposes also to the most toxic OH.

Because of the large energy gain of the
reduction of OH to H2O, this radical
reacts
instantaneously
with
any
biological molecule in its immediate
environment by abstracting hydrogen
atom.
29

Production of ROS
in endothelium and neutrophils
OH

OONO-

l-arginine
e
n
d
o
t
h
e
l
i
u
m

O2
ATP

NADPH

XDH

NO

AMP adenosine

inosine

hypoxanthine
O2
XO
uric acid O2OH

NO

NOS

H 2 O2

O2

NADPH
NADPH
oxidase

O2H2O2

OH

NADP+

MPO
HOCl

MPO

n
e
u
t
r
o
p
h
i
l
30

Institute of Experimental Pharmacology, SASc, Bratislava, SK

Effects of ROS on the endothelium and development

of atherosclerosis
atherosclerotic lesion
recruitment of
macrophages

cell proliferation

release of growth factor

active oxygen,
collagenase, elastase,
lipases, proteases

Plasma

adherence
of platelets
endothelial cells

Monocyte

Membrane LDL
damage

Intima
Fatty Streak

activated
oxygen
iron/copper

Oxidatively modified LDL

apoB-bound 4-hydoxynonenal, oxidized lipids,

fatty acid hydroperoxides

Foam
cells

Tissue
macrophages

Monocyte

31
based on J.P. Kehrer (1993)

Diseases that may have ROS related pathogenesis

Airways

Normobaric hyperoxic injury

Bronchopulmonary dysplasia
Idiopathic pulmonary fibrosis
Respiratory distress syndromes (ARDS, IRDS)
Emphysema
Chronic bronchitis & asthma bronchiale
Asbestosis
Inhaled pollutants, smoke, chemicals (e.g. paraquat,
bleomycin) & oxidants (e.g. SO2, NOx, O3)

Gut

Ischemia/reperfusion
Crohns disease
Ulcerative colitis & necrotizing enterocolitis
Gastric & intestinal ulcers
Chemicals (e.g. NSAID)

32

II
Heart and vessels

Ischemia/reperfusion (after infarction, transplantation)

Chemicals (e.g. ethanol, doxorubicin)
Atherosclerosis/hypertension
Selenium deficiency
Vasculitis

Brain and nerves

Hyperbaric hyperoxic injury

Parkinson's disease
Alzheimers disease (details see in the next panel)
Amyotrophic lateral scleroses
Neuropathies (e.g. diabetic)
Neurotoxins (e.g. 6-hydroxydopamine, MPTP)
Vitamin E deficiency
Neuronal ceroid lipofuscinoses
Traumatic injury/hemorrhage/inflammation
Ischemia/reperfusion
HIV-dementia
Multiple sclerosis

33

III
Blood

Chemicals (e.g. phenylhydrazine, primaquine, sulphonamides,

lead)
Protoporphyrine photooxidation
Malaria
Anemias (sickle cell, favism)

Liver

Ischemia/reperfusion
Chemicals (e.g. halogenated hydrocarbons, quinones, ethanol,
acetaminophen)
Accumulation of iron or copper
Endotoxin

Kidney

Autoimmune nephrosis (inflammation, e.g.

glomerulonephritis)
Chemicals (e.g. aminoglycosides, heavy metals)

34

IV
Pancreas
Acute & chronic pancreatitis
Diabetes mellitus

Eye
Retinopathy of prematurity
Photic retinopathy
Cataracts
Laser photoablation

Skin
Radiation (solar, ionising)
Thermal injury
Chemicals (photosensitizers, e.g. tetracyclines)
Contact dermatitis
Porphyria

35

V
Muscle

Muscular dystrophy
Multiple sclerosis
Exercise

Others
Aging
Pregnancy and newborn complications
Radiation injury
Cancer
Chemicals (e.g. alloxan, iron overload, radiosensitizers)
Autoimmune diseases (e.g. rheumatoid arthritis, lupus
erythematodes)
Inflammation (in general)
36

Potential antioxidant therapy I

Inhibitors

of ROS synthesis

NADPH-oxidase Inhibitors
Flavoprotein inhibitors
(FAD analogs, antibodies of cytP450 reductase)
Agents forming complexes with Fe2+ in cyt b
(butylisocyanide, imidazole, pyridine)
Mg2+(enabling FAD binding),Fe2+ ,Cu2+ chelators
(bathophenantroline, EDTA, EGTA, deferoxamine,
bilirubin)
Thiol reagents (N-ethylmaleimide, 1-naphtol, 1,4naphtoquinone)
NADPH analogs (NADPH 2,3-dialdehyde)
Inhibitors of metabolism of AA and PLA2
IMAO (Deprenyl)
Others (corticosteroids, diphenyliodonium)
Inhibitors of xanthine oxidase (tungsten, oxypurinol,
allopurinol, pterinaldehyde, folic acid)
Antibodies against leukocytes

37

II
Agents

supporting and complementing enzymatic

protective systems

Superoxide

dismutase (SOD)
SOD (Lip-SOD,PEG-SOD)
Copper diisopropylsalicylate

SOD mimetics
Catalase (Cat)
Cat (Lip-CatTP, Peg-CatTP)
Glutathionperoxidase (GTPx)
GSH, GSH methylester, GSH diethylmaleate
Low m.w. thiols (e.g. cystein)
High m.w. thiols (e.g. albumin)
L-2-oxothiazidolidine-4-carboxylate
N-acetylcysteine
Ebselen
Selenium
Lactoperoxidase & DT-diaphorase

38

III

Drugs interfering with iron and copper metabolism

(deferoxamine, hemopexine, ferritin, transferrin, lactoferrin,

ceruloplasmin, serum albumin)

Antioxidants

Vitamins and their analogues (vitamin E, vitamin C, carotenoids,

oxycarotenoids)
Phenol derivatives (eugenol, guajacol, probucol, N,N-diphenylphenylendiamine)
Flavone derivatives (flavonoids, isoflavonoids, allirazine, green tea)
Indol derivatives (stobadine, carvedilol, melatonin, -carbolines)
Xanthine derivatives (allopurinol, oxypurinol, uric acid)
21-amino steroids (lazaroids)
Antiinflammatory drugs (piroxicam, flufenamic acid, mefenamic
acid, hydroquinone, sulindac, fenylbutazone, indomethacin,
ibuprofen, naproxen, levamisole, sulfasalazine, acetylsalicylic
acid)
Hypolipidemics (lovastatin)
Proteins (albumin)
39

IV
Agents

containing sulfur (cysteine, cysteamine, GSH,

dithiothreitol, N-acetylcysteine, ACE inhibitors,
dimethylthiourea, thiourea, thiomalate, hypotaurine, taurine,
penicillamine, 2-amino-2-thiazole, dihydrolipoate,
-mercaptopropionyl glycine, N-2-mercaptopropionyl glycine,
-mercaptoethanole, D,L-methionine, other low and high m.w.
thiols)
Nitroso compounds ( .NO, nitrosopine)
Other drugs ( -adrenolytics, H2-antihistaminics, calcium channel
blockers, pentoxyphylline, carbanilates, urea, bilirubin, glucans,
manitol, glucose, 2-methylaminochromans, DMSO, BHT, BHA,
2-MEA, etoxiquin, -lipoic acid, Zn2+)
40

V
Inhibition

of O2.- formation

Nonsteroid antiflogistics

Antiasthmatics
( adrenomimetics, corticoids, methylxanthines)
Prostaglandins
Flavonoids
Antibiotics
(e.g. minocycline)
Antimalarics
Inhibitors of ACE
Dipyridamol

41

VI/a
Scavenging
Scavenging

or removal of ROS
of generated O2.-

Scavenging

HOCl

Scavenging

or quenching of 1O2

Flavonoids & other natural products

Vitamins E, C, A( -carotene)
Synthetic analogs of PGB2
Dipyridamol
Pentoxiphylline
Antibiotics
.NO donors
5-acetylsalicylic acid
Uric acid
Uric acid
Taurine, hypotaurine
Silymarine
-carotene
Vitamin E
Stobadine

42

VI/b
Scavenging
Removal

or removal of ROS

of H2O2

Catalase (not working in the presence of .NO)

N-acetylcysteine

Elimination

of OH.

Manitol
Thiourea
Stobadine
Melatonin
Probucol
5-acetylsalicylic acid
Lazaroids
DMSO, DMTU, BHT
Uric acid
Glucose
43

 Scavenging

VI/c

or removal of ROS
Lipid oxidation chain breaking antioxidants
(anti LO. and LOO.)
Bilirubin
Vitamins E
Vitamin C
-carotenoids and oxycarotenoids
Stobadine
Melatonin
-lipoic acid
Uric acid
Lazaroids
BHT, BHA
Ehoxyquin
2-methylaminochroman

Some positive results from preclinical and clinical studies with the
pyridoindole STOBADINE, which possesses significant antioxidant,
mainly hydroxyl radical scavenging, lipid oxidation chain breaking
and singlet oxygen quenching properties, as an example, are 44
presented in the following panels.

% of Max. Relax.

Protection by STOBADINE (STB) of the acetylcholine

induced relaxation in rat aortic rings
caused by reversible occlusion of aorta in vivo (I/R)
H

H3C

35
30
25
20
15
10
5
0

CH3

N
H

.2HCl

Sham
I ischemia; R reperfusion
P<0.05 I/R vs Sham
P<0.05 I/R+STB vs I/R

I/R

I/R + STB

STOBADINE
45

Institute of Experimental Pharmacology, SASc, Bratislava, SK

STOBADINE (STB) effect on experimetal

myocardial infarction (MI) in dogs

MI area %

25
20

15

10
5

0
C

STB

3hr occlusion of the

anterior descendent
branch of the left
coronary artery
Stobadine (1 mg/kg iv)
given 30 min after the
occlusion
Reduction of the
infarction area by 28%
( P < 0.05)

46SK
Institute of Experimental Pharmacology, SASc, Bratislava,

STOBADINE effect on transmission in rat

hippocampal slices during hypoxia/reoxygenation

A-control , B-hypoxia, C- reoxygenation in untreated slices,

D-reoxygenation in stobadine treated slices

47
Institute of Experimental Pharmacology, SASc, Bratislava, SK

Antianginal effect of STOBADINE (STB)

Phase II clinical study

N
um
ber/ week

8
6
4
2
0

Attacks
Nitroglycerine tbl.
STB

-1

*
*

*
*

3 4
w eeks

Patients with angina

pectoris (stable and effort)
(n = 13)
Effect of 4 week treatment
with STOBADINE (up to
100 mg/day p.o.)
Significant decrease in the
No. of anginal attacks
Significant (*
( P<0.05)
decrease in the No. of
selfadministered
nitroglycerine tablets
48

Institute of Experimental Pharmacology, SASc, Bratislava, SK

Therapeutic relevance of the use of antioxidants

DISEASE
ANTIOXIDANT THERAPEUTIC
I
SUCCESS
Cardiovascular

Carotenoids
Ascorbic acid
Tocopherols
Selen
Probucol
Flavonoids

Newborn hypoxia induced

injuries

Penicillamine
Tocopherols

+
+

Ischemia/Reperfusion
(of the heart, brain,
gut, kidney)

SOD, SOD+CAT
Lipoic acid
Allopurinol
Tocopherols
Deferoxamine

Transplantation and
tissue preservation

SOD, SOD+CAT
Tocopherols
Ascorbic acid

+
+
+
+
+

49

II

DISEASE

ANTIOXIDANT THERAPEUTIC
SUCCESS

Intravascular hemorrhage

Tocopherols
Ascorbic acid

+
+

Platelets aggregation

Flavonoids
Stobadine

+
+

Hemochromatosis

Deferoxamine

++

Head trauma
(details see in the next

Lazaroids
Stobadine derivatives
Phenyl-butyl-nitones

+
+

Subarrachnoidal
hemorrhage

Lazaroids

Respiratory distress
syndromes (IRDS, ARDS)

SOD
Allopurinol
Tocopherols

+
+
+

Bronchial asthma

SOD+CAT
Thiols
Tocopherols

panel)

50

III

DISEASE

ANTIOXIDANT THERAPEUTIC
SUCCESS

Flu (cold)

Ascorbic acid

Retrolental fibroplasia

Tocopherols

Cataract

Tocopherols

Inflammatory diseases of the 5-aminosalicylates

gut (IBD)
Sulfasalazine
Sulfapyridine
SOD+CAT
Glucans

+
+
+

Hepatopathies

Lipoic acid
Silymarin
Stobadine

Paracetamol intoxication

N-acetylcysteine

+
++

Chemical poisonings

Glutathione
Deferoxamine

+
+

Photosensibilization

Carotenoids
Tretionin

++
+

51

IV

DISEASE

ANTIOXIDANT THERAPEUTIC
SUCCESS

Rheumatoid arthritis

SOD
Penicillamine
Deferoxamine

Parkinsomism

Tocopherols

Wilsons disease

Penicillamine

++

Cerebro-vascular spasms

Carotenoids
Tocopherols
Thiols

Cancer

SOD+CAT
Carotenoids
Ascorbic acid
Tocopherols
Selen
Flavonoids
Thiols

52

ROS in the sequence of events in

NEUROTRAUMA
TRAUMA
Excitatory aminoacid release (GLU)
Ca2+ influx into the cells
Activ. of inflam. cascade
Protease/lipase activation
(PAF, eikosanoids,
Cell depolarization
cytokines, PMN activ.)
( Mg block of NMDA rec.)
ROS generation
Na+influx
cell devastation
Edema
NEURONAL DEATH
TRIAD : EXCITOTOXICITY, Ca-OVERLOAD, OXIDATIVE STRESS

Therapeutic interventions: -SH donors (N-acetylcysteine), lazaroids,

steroids, deferoxamine, SOD, vitamines A,E,C, pyridoindoles,
stobadine, PBN, flavonoids (quercetine), PAF antag. (BN 520210)

53

Conclusions I

ROS act by:

increase of membrane lipid peroxidation
increase of prostaglandin production
increase of intracellular free calcium
alteration of conductivity of ion channels
alteration of enzyme activity
alteration of release/action of neurotransmitters
reduction of half-life of biologically active
substances

damage of proteins

damage of DNA, genes and protein synthesis

damage of carbohydrates
Mechanisms of ROS action differ in various biological tissues.
Their actions depend upon condition of the tissue itself, the 54
corresponding epithelium, endothelium, innervation, etc.

Conclusions II
The effects of ROS could be prevented or
stopped by:

reduction of their generation

- elimination of undesirable physical and chemical

influences
- protection of tissues from chronic inflammation
- protection of tissues from ischemia

their elimination

interaction with their effects

- substitution with antioxidant enzymes

- substitution with non-enzyme antioxidants
and scavengers
- protection of cells from intracellular free
calcium accumulation and its effects

55

Conclusions III
Therapeutic success with the use of
antioxidants, quenchers and scavengers
There are

Promising clinical results

e.g. in photosensibilization, paracetamol intoxication,

hemochromatosis

Controversial clinical results

e.g. in ischemia/reperfusion, subarachnoidal hemorrhage,

respiratory distress syndromes

Minimal therapeutic effects

e.g. in asthma bronchiale, cancer

56

Thank you
Matur nuwun

57