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THE ORALLY EFFECTIVE MIXTURE OF SOD AND GLIADIN GliSODin® PROTECTS AGAINST OXIDATIVE DNA DAMAGE

Authors:

Abstract

bounded (tail moment from 0.08 (0.06-0.12) to 0.11 (0.07-0.23), p=0.020) the otherwise marked increase (Vehicle: tail moment from 0.11 (0.09-0.13) to 0.43 (0.40-0.73), p=0.063) in DNA strand breaks after HBO exposure (p=0.005 GliSODin® vs. Vehicle after HBO). GliSODin® also reduced oxidative DNA damage related to surgical stress and ischemia-reperfusion after aortic clamping (tail moment from 0.09 (0.08-0.11) to 0.12 (0.11-0.14), p= 0.023) in comparison to Vehicle (tail moment from 0.12 (0.11-0.15) to 0.48 (0.35-0.60). p=0.031) resulting in a statistically significant intergroup difference 2 hrs after declamping (p= 0.021, GliSODin® vs. Vehicle). While there was no intergroup difference in the baseline values of isoprostane levels (p=0.391), these values significantly increased both in hepatic venous (p=0.003) and portal venous samples (0.006). No such effect was observed in GliSODin® group. Neither SOD nor catalase activities were significantly affected by GliSODin® ingestion (baseline values GliSODin® vs. Vehicle, p= 0.289 ) or by stress related to surgery and aortic cross clamping (GliSODin® vs. Vehicle p= 0.312).
THE ORALLY EFFECTIVE MIXTURE OF SOD AND GLIADIN
GliSODin
®
PROTECTS AGAINST OXIDATIVE DNA DAMAGE
Background:
Generation of oxygen-derived radicals has been demonstrated to be the major mechanismof ischemia-reperfusion induced damage [1]. Reactive oxygen
species interact with DNA leading to structural alteration [2]. Recently, we showed that GliSODin
®
, an orally effective mixture of SOD and wheat gliadin,
protected against hyperbaric oxygen (HBO)-induced DNA damage assessed by single cell gel electrophoresis (comet assay) [4]. Since DNA damage was less
marked than in previous studies [3], we investigated whether GliSODin
®
is also protective when during more severe oxidative stress.
Materials and Methods:
After 2 weeks of feeding once a day with 1250 IU of GliSODin
®
(n = 9) or vehicle (n = 5) blood samples were taken from swine (body weight 50 (47-53) kg). DNA
damage (tail moment in the alkaline version of the comet assay) was evaluated in isolated lymphocytes (Ficoll gradient) before and after HBO exposure (2 hrs
at 4 bar O
2
) (GliSODin
®
n=9, Vehicle n=5). Comet assay was performed as well in whole blood samples taken at different time points (before and after 30
minutes thoracic aortic cross-clamping as well as 2 and 4 hours after declamping) during thoracic aortic surgery (GliSODin
®
n=8, Vehicle n =7). Plasma 8-
isoprostane (8-epi Prostaglandin F
2
) concentrations as a direct marker of lipid peroxidation were determined using an enzyme immunoassay kit (Cayman
Chemicals, Ann Arbor, MI) in portal (PV) and hepatic venous (HV) blood before and after aortic cross clamping. Antioxidant enzyme SOD (RANSOD kit, Randox
Laboratories Ltd, U.K.) and catalaseactivity (assayed by a method in which the disappearance of peroxide is followed spectrophotometrically) were assessed
on whole blood samples taken during aortic cross clamping and onlymphocytes isolated before and after HBO exposure. Data are median (range). After
exclusion of normal distribution data using a Kolmogorov -Smirnov-test, time dependent differences within groups during aortic surgery were analyzed with a
Friedman repeated measures ANOVA and, if appropriate, by a Dunns test. Differences before and after HBO exposure were analyzed with a Wilcoxon signed
rank test. Intergroup differences were analyzed with Mann-Whitney rank sum test.
Results
There was no difference in DNA damage before exposure to HBO (p=0.255 GliSODin
®
vs. Vehicle). GliSODin
®
bounded (tail moment from 0.08 (0.06-0.12) to
0.11 (0.07-0.23), p=0.020) the otherwise marked increase (Vehicle: tail moment from 0.11 (0.09-0.13) to 0.43 (0.40-0.73), p=0.063) in DNA strand breaks after
HBO exposure (p=0.005 GliSODin
®
vs. Vehicle after HBO).
GliSODin
®
also reduced oxidative DNA damage related to surgical stress andischemia-reperfusion after aortic clamping (tail moment from 0.09 (0.08-0.11) to
0.12 (0.11-0.14), p= 0.023) in comparison to Vehicle (tail moment from 0.12(0.11-0.15) to 0.48 (0.35-0.60). p=0.031) resulting in a statistically significant
intergroupdifference 2 hrs after declamping (p= 0.021, GliSODin
®
vs. Vehicle). While there was no intergroup difference in the baseline values of isoprostane
levels (p=0.391), these values significantly increased both in hepatic venous (p=0.003) and portal venous samples (0.006). No such effect was observed in
GliSODin
®
group. Neither SOD nor catalaseactivities were significantly affected by GliSODin
®
ingestion (baseline values GliSODin
®
vs. Vehicle, p= 0.289 ) or by
stress related to surgery and aortic cross clamping (GliSODin
®
vs. Vehicle p= 0.312).
COMET ASSAY CLAMPING
Start of
surgery
Before
clamping
Before
declamping
2 hrs after
declamping
COMET ASSAY LYMPHOCYTES IN HBO
Before
HBO
After
HBO
Tail moment
Ethidiumbromide-
stained nuclei after
performance of the
comet assay.
A)nucleus without
detectable DNA
damage,
B) severely
damaged nucleus with
pronounced ,"comet
tail".
*: before clamping vs2 hrs after declamping
Conclusions:
Pretreatment with the new nutritional formula of SOD-wheat gliadin (GliSODin
) allows to prevent oxidative DNA damage related to HBO treatment or
ischemia-reperfusion injury.
The unaltered SOD activities after oral SOD ingestion are probably due to relatively low SOD supplementation when compared to total blood SOD pool
[3,5]. The effect of SOD, thus, most likely results from an immune response and through a nitric oxide dependent mechanism [5].
References:
1
MacCord JM. Oxygen-derived free radicals in postischemictissue injury. N Eng J Med 1985; 312:159-163
2
Cooke MS, Evans MD, Dizdaroglu M, Lunec J Oxidative DNA damage: mechanisms, mutation, and disease. FASEB J 2003; 17:1195-1214
3
Muth CM, GlenzY, Klaus M, RadermacherP, Speit G, Leverve XM. Influence of an orally effective mixture of SOD and wheat gliadinon
hyperbaricoxygen-related cell damage. Free RadicRes 2004;38:927-932
4
Speit G, DennogC, Radermacher P, Rothfuss A. Genotoxicityof hyperbaric oxygen. MutatRes2002;512:111-119
5
Vouldoukis I, LacnaD, Kamate C, CosteP, CalendaA, Mazier D, Conti M, Dugas B. Antioxidant and antiinflammatory
properties of a Cucumis meloLC extract rich in superoxide dismutase activity. J Enthnopharmacol2004;94:67-75
* p<0.05 vs baseline. # p<0.05 vs Vehicle
* p<0.05 vs baseline. # p<0.05 vs Vehicle
2091
(2012-2148)
2264*
(1946-2691)
1853
(1568-2028)
Vehicle
1958
(1428-2604)
2339
(1990-2637)
2309
(1700-2419)
GliSODin
PORTAL
VEIN
1842
(1655-1999)
2188*
(1837-2645)
1762
(1528-1898)
Vehicle
1716
(1356-1993)
1797
(1540-2203)
1945
(1289-2097)
GliSODin
HEPATIC
VEIN
4 hrs after
declamping
2 hrs after
declamping
before
clamping
8-ISOPROSTANE
pg / g protein
86
(72-87)
80
(71-87)
82
(74-86)
Vehicle
81
(77-87)
83
(79-92)
80
(77-88)
GliSODin
Catalase
[kU/gHb]
1.4
(1.3-1.6)
1.5
(1.4-1.7)
1.4
(1.3-1.5)
Vehicle
1.6
(1.3-1.8)
1.6
(1.4-1.7)
1.6
(1.3-1.7)
GliSODin
SOD
[U/gHb]
4 hrs after
declamping
2 hrs after
declamping
before
clamping
4.2
(3.8-4.6)
2.8
(2.3-3.5)
1.3
(1.1-1.5)
Vehicle
3.9
(3.5-4.3)
2.7
(2.5-3.2)
1.2
(1-1.3)
GliSODin
After
HBO
4.2
(3.8-4.8)
3.1
(2.5-3.8)
1.3
(1.0-1.4)
Vehicle
3.4
(3.2-3.8)
2.2
(1.9-2.8)
1.2
(0.7-1.3)
GliSODin
Before
HBO
SODt
[U/gHb]
SOD CuZn
[U/gHb]
SOD Mn
[U/gHb]
M. Albicini
1,4
, J. Kick
2
, B. Hauser
1,5
, U. Ehrmann
1
, X. Leverve
6
, P. Radermacher
1
, G. Speit
3
, C.M. Muth
1,3
1
Sektion Anästhesiologische Pathophysiologieu. Verfahrensentwicklung,
2
Abt. Thorax- und Gefäßchirurgie,
3
Abt. Humangenetik,
Universitätsklinikum, Ulm, Germany;
4
Istituto di Anestesiologiae Rianimazione dell'Universitadegli Studi di Milano: AziendaOspedaliera, Polo UniversitarioSan Paolo, Milano, Italy
5
Aneszteziológiai és IntenzívTerápiás Klinika, Semmelweis Egyetem, Budapest, Hungary.
6
Laboratoire de Bioénergétique Fondamentaleet Appliquée, Université Joseph Fourier, Grenoble, France;
B. Hauser was supportedby the
Alexander von HumboldtFoundation
... In addition, this SOD-gliadin complex has been shown to • Inhibit oxidative stress (Albicini et al., 2005;Nakajima et al., 2009;Vouldoukis et al., 2004) • Inhibit ultraviolet oxidative stress (Mac-Mary, Sainthillier, Courderotmasuyer, Creidi, & Humbert, 2007) • Promote immune modulation (Okada et al., 2006) • Inhibit vascular inflammation Results of a few studies (Arent, DiFabio, Greenwood, Pellegrino, & Williams, 2005;Hong, Hong, Chang, & Cho, 2004) suggested that the melon extract (Cucumis melo LC) may attenuate peroxidative and inflammatory response to exercise in athletes. ...
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Article
Hyperbaric oxygen (HBO) treatment is applied as a therapy for a wide variety of diseases with symptoms caused by lack of oxygen in the target tissues. However, it is known that exposure to high concentrations of oxygen may lead to oxidative stress and cause cell and tissue damage. Oxygen toxicity and possible cancer-promoting effects of HBO therapy have been a matter of serious concern. Although a cancer-inducing effect of HBO was not found to date, recent studies clearly indicated an induction of oxidative DNA damage in blood cells of healthy subjects after HBO under therapeutic conditions. The biological significance of this finding has been investigated in a series of in vitro and in vivo tests. This review summarizes these studies and critically discusses potential adverse genetic effects of HBO therapy. Furthermore, since an induction of anti-oxidative defense mechanisms has been determined after HBO exposure, a modified treatment regimen of HBO therapy is proposed which avoids genotoxic effects.