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HY DRO GEN-RICH PBS PRO TECTS CUL TURED HU MAN
CELLS FROM ION IZ ING RA DI A TION-IN DUCED
CEL LU LAR DAM AGE
by
Liren QIAN 1, Bailong LI 1, Fei CAO, Yuecheng HUANG, Shulin LIU,
Jianming CAI*, and Fu GAO*
De part ment of Ra di a tion Med i cine, Fac ulty of Na val Med i cine,
Sec ond Mil i tary Med i cal Uni ver sity, Shang hai, P. R. China
Sci en tific pa per
UDC: 614.8.086.5:66.094.1
DOI: 10.2298/NTRP1001023Q
Hydroxyl rad i cals play an im por tant role in ion iz ing ra di a tion-in duced cel lu lar dam age, while
hy dro gen can se lec tively re duce hydroxyl rad i cals in vi tro. This study was de signed to test the
hy poth e sis that hy dro gen-rich PBS may be an ef fec tive radioprotective agent in vi tro. Com -
pared to cells pretreated with out hy dro gen, we dem on strated that treat ing cells with hy dro -
gen-rich PBS be fore ir ra di a tion could sig nif i cantly in hibit IR-in duced apoptosis, in crease vi -
a bil ity of human in tes ti nal crypt cells, sig nif i cantly in crease en dog e nous an ti ox i dant, and
de crease malondialdehyde and 8-hydroxydeoxyguanosine con cen tra tions of human lym pho -
cyte AHH-1 cells. It is con cluded that hy dro gen has a po ten tial as an ef fec tive and safe
radioprotective agent.
Key words: hy dro gen, ionizing ra di a tion, radioprotection, apoptosis, re ac tive ox y gen spe cies
IN TRO DUC TION
Hy dro gen is the most abun dant chem i cal el e -
ment. It is a col or less, odor less, non-me tal lic, taste -
less, highly flam ma ble di atomic gas which was con -
sid ered as a physiological in ert gas. Hy dro gen is
sel dom re garded as an im por tant agent in med i cal us -
age. Howerver, Ohsawa et al. [1] found that mo lec u -
lar hy dro gen could se lec tively re duce cytotoxic re ac -
tive ox y gen spe cies, such as ·OH and ONOO– in vi tro
and ex ert ther a peu tic an ti ox i dant ac tiv ity in a rat mid -
dle ce re bral ar tery oc clu sion model.
Re ac tive ox y gen spe cies (ROS) or re ac tive ni -
tro gen spe cies (RNS), such as hydroxyl rad i cal (·OH),
superoxide an ion (O2-), hy dro gen di ox ide (H2O2), ni -
tric ox ide (NO), peroxynitrite (ONOO-), ap pear to
play a crit i cal role in ce re bral, myo car dial and hepatic
ischemia-reperfusion injuries, trans plan ta tion in ju -
ries, and other in ju ries [2-4]. It has also been dem on -
strated that H2 is ef fec tive in the prevention of these in -
ju ries. How ever, the po ten tial ef fect of hy dro gen gas
on an other dam age type in which free radicals play an
im por tant role is largely ig nored. That type is the dam -
age in duced by ir ra di a tion.
Approximately 65% of the DNA dam age is
caused by the in di rect ef fect of free rad i cals, such as
hydroxyl rad i cals (·OH), that are formed from the
radiolysis of surrounding wa ter mol e cules and that
suc ces sively at tack DNA [5]. Lipid peroxidation
(LPO) is also con sid ered as a crit i cal event dur ing ion -
iz ing ra di a tion in duced dam age [6]. Apart from ge -
netic dam age and lipid peroxidation, ROS can also al -
ter the bal ance of en dog e nous pro tec tive sys tems such
as glutathione and en zy mic an ti ox i dant de fence sys -
tems [7] . The en dog e nous an ti ox i dant defences are in -
ad e quate to re duce the ra di a tion-in duced free rad i cal
changes. Ap pro pri ate an ti ox i dant in ter ven tion seems
to in hibit or re duce free rad i cal tox ic ity and thus of fers
pro tec tion against ra di a tion.
There fore, we rea soned that hydrogen might be
pro tec tive against det ri men tal ef fects of ra di a tion.
How ever, the ap pli ca tion of H2 gas in ha la tion is not
con ve nient and may be dan ger ous be cause the gas is
in flam ma ble and ex plo sive. On the other hand, H2 gas
sat u rated PBS, which is called hy dro gen-rich PBS, is
easy to ap ply and safe. In the cur rent study, we dem on -
strated that hy dro gen-rich PBS treat ment can pro tect
human cells from g-ra di a tion in vi tro.
L. Qian, et al.: Hy dro gen-Rich PBS Pro tects Cul tured Hu man Cells from ...
Nu clear Tech nol ogy & Ra di a tion Pro tec tion: Year 2010, Vol. 25, No. 1, pp. 23-29 23
*Cor re spond ing au thors; e-mails: cjm882003@ya hoo.com.cn
(J. Cai); gaofu_2002@ya hoo.com.cn (F. Gao)
1L. Qian and B. Li con trib ute equally to this paper
MA TE RI ALS AND METH ODS
Prep a ra tion of hy dro gen-rich PBS
Hy dro gen was be ing dis solved in PBS for 6
hours un der high pres sure (0.4 MPa) in or der to reach a
su per sat u rated level by us ing hy dro gen-rich wa -
ter-pro duc ing ap pa ra tus which was pro duced by our
de part ment. The sat u rated hy dro gen PBS was stored
un der the at mo spheric pres sure at 4 °C in an alu mi num
bag with no dead vol ume. Hy dro gen-rich PBS was
freshly pre pared ev ery week, which en sured that a
con cen tra tion of more than 0.6 mmol/L was main -
tained. Gas chro ma tog ra phy was used to con firm the
con tent of hy dro gen in PBS by the method de scribed
by Ohsawa, et al. [1].
Hy dro gen treat ment of cul tured cells
Hu man lym pho cyte AHH-1 cells and in tes ti nal
crypt HIEC cells were main tained in RPMI 1640
(Invitrogen) with 10% fe tal bo vine se rum and 1% pen -
i cil lin-strep to my cin-glutamine at 37 °C in a 5% CO2
hu mid i fied cham ber. For radioprotective stud ies, cells
were treated with different vol ume of hy dro gen-rich
PBS and ac cord ingly we added dif fer ent vol ume of
PBS in or der to ob tain the de sired con cen tra tion of H2
and make the fi nal vol ume of the me dium the same.
Then, the treated cells were im me di ately ir ra di ated
with dif fer ent doses of g-ray, de pend ing on the re -
quire ment of the pres ent study.
Ir ra di a tion
Co balt-60 gamma rays in irradiation cen ter
(Fac ulty of Na val Med i cine, Sec ond Mil i tary Med i cal
Uni ver sity, China) were used for the ir ra di a tion pur -
pose. Cells (with or with out hy dro gen pre-treat ment)
were ex posed to dif fer ent doses of ra di a tion, de pend -
ing on the re quire ment of the pres ent study.
Clonogenic sur vival
Col ony-form ing as say was per formed as pre vi -
ously de scribed [8]. Briefly, cal cu lated num bers of
cells were plated to en able nor mal iza tion for plat ing
ef fi cien cies. Pretreated cells were then ir ra di ated with
0, 2, 4, 6, or 8 Gy. Af ter in cu bat ing for 7 days, the
plates were fixed with 70 % EtOH and stained with 1%
meth y lene blue. Col o nies con sist ing of >50 cells were
counted un der mi cro scope. The sur vival frac tions
were cal cu lated as (num ber of col o nies / num ber of
cells plated) / (num ber of col o nies for cor re spond ing
con trol / num ber of cells plated).
Lac tate dehydrogenase (LDH) leak age as say
LDH leak age as say was car ried out us ing LDH
cytotoxicity de tec tion kit (Nanjing KeyGen Biotech.
Co. Ltd.) ac cord ing to the pro to col in the user’s man -
ual. Cells were pretreated with hy dro gen-rich PBS and
the fi nal con cen tra tion of H2 was main tained above
0.3 mmol/L. The cells were immediately ex posed to
gamma ra di a tion and then trans ported to an ice bucket.
Af ter 4 hour time pe riod we ana lysed the con tent of
LDH in cell sus pen sion.
Apoptosis as says for cul tured cells
Apoptosis was de ter mined by Annexin V-APC
and propidium io dide stain ing us ing apoptosis de tec -
tion kit (Bipec Biopharma). Treated cells were in cu -
bated with Annexin V-APC for 15 min utes at 4 °C and
propidium io dide for 5 min utes at room tem per a ture.
The cells were then an a lyzed by flow cytometry. Al ter -
na tively, apoptosis was de ter mined by Hochest33258,
flourescein diacetate (FDA) and propidium io dide
stain ing. The treated cells were washed with PBS
twice, and then stained with 40 mg/L flourescein
diacetate, 20 mg/L Hoechst33258 at room tem per a ture
for 15 min utes, and stained with 20 mg/L propidine io -
dine at room tem per a ture for 5 minutes. The cel lu lar
mor phol ogy was ob served us ing Olym pus BX60 flu o -
res cent mi cro scope equipped with Retiga 2000R dig i tal
cam era. The av er age per cent age of apoptotic cells was
cal cu lated in 5-7 ran domly se lected high power fields
(HPF).
De ter mi na tion of malondialdehyde (MDA)
superoxide (SOD) glutathione (GSH)
MDA is a break down prod uct of the ox i da tive
deg ra da tion of cell mem brane lipids and is gen er ally
con sid ered an in di ca tor of lipid peroxidation. SOD is a
scav en ger of superoxide, and GSH is an im por tant cel -
lu lar non-en zy matic an ti ox i dant. In the pres ent study,
4 hours after trans porting the ir ra di ated cells to an ice
bucket, the con cen tra tions of MDA, SOD, and GSH
were mea sured, re spec tively, by us ing the MDA,
SOD, GSH as say kit (Nanjing KeyGen Biotech. Co.
Ltd.) ac cord ing to the pro to cols in the user’s man ual.
De ter mi na tion of 8-OHdG con cen tra tion
Half an hour af ter the ir ra di a tion, the 8-OHdG
con cen tra tion was mea sured by us ing hu man 8-OHdG
elisa kit (Nanjing KeyGen Biotech. Co. Ltd.) ac cord -
ing to the pro to col in the user’s man ual. Briefly af ter
that, the treated cells were lysed by a cell lysis buffer.
Af ter the cen tri fuge, we added sus pen sion to plate
L. Qian, et al.: Hy dro gen-Rich PBS Pro tects Cul tured Hu man Cells from ...
24 Nu clear Tech nol ogy & Ra di a tion Pro tec tion: Year 2010, Vol. 25, No. 1, pp. 23-29
wells coated with hu man 8-OHdG an ti body, and se -
quen tially treated them with biotinylated anti-lgG and
streptavidin-HRP. Af ter that, we added TMB sub strate
so lu tion, and TMB sub strate changed color into blue at
HRP en zyme-cat a lyzed. At the ef fect of acid, the color
fi nally be come yel low. The in ten sity of this col ored
prod uct is di rectly pro por tional to the con cen tra tion of
8-OHdG. Mea sur ing the op ti cal den sity (OD) at 450
nm with a microtiter plate reader, we cal cu lated hu man
8-OHdG con cen tra tion by the stan dard curve.
STA TIS TI CAL ANAL Y SIS
Data are ex pressed as means ±S. E. M. (stan dard
er ror of the mean) for each ex per i ment. The num ber of
sam ples is in di cated in the de scrip tion of each ex per i -
ment. Sta tis ti cal anal y sis was per formed by us ing one
way anal y sis of vari ance. Be tween groups, vari ance
was de ter mined us ing the Stu dent-Newman-Keuls post
hoc test. A P value of less than 0.05 was con sid ered to be
sta tis ti cally sig nif i cant.
RE SULTS
Hy dro gen-rich PBS in creases clonogenic
sur vival of ir ra di ated HIEC cells
To study radioprotective ef fects of H2 in a cell cul -
ture, we ex am ined the vi a bil ity of the ir ra di ated hu man
in tes ti nal crypt (HIEC) cells us ing a clonogenic sur -
vival as say. Pre treat ment of HIEC cells with 0.1-0.4
mmol/L H2 be fore ir ra di a tion sig nif i cantly in creased
cell sur vival as com pared to the cells treated with ra di a -
tion alone at all ex am ined doses (up to 8 Gy), fig. 1(a).
Hy dro gen-rich PBS de crease cel lu lar lactate
dehydrogenase (LDH) leak age of ir ra di ated
HIEC cells
Be side the cell vi a bil ity, we also determined
LDH ac tiv i ties to es ti mate cel lu lar LDH leak age from
dam aged cells. The re sult in di cated that pre treat ment
with 0.3 mM H2 be fore ir ra di a tion sig nif i cantly de -
creased LDH leak age of HIEC cells which were ex -
posed to dif fer ent doses of g-ra di a tion, fig. 1(b). This
re sult too was con sis tent with the re sult ob tained by
cell vi a bil ity ob ser va tion.
Hy dro gen-rich PBS at tenu ates apoptosis in
ir ra di ated HIEC cells
To de ter mine the ra di a tion-in duced apoptosis of
the ir ra di ated HIEC cells, we an a lyzed the treated cells
by us ing Annexin V-APC and propidium io dide stain -
ing in flow cytometry as say. The early apoptotic cells
de creased when pretreated with hy dro gen-rich PBS as
com pared to the cells pretreated with PBS, fig. 2(a)
and (b), 10.2% vs. 21.5%, re spec tively). We fur ther
eval u ated the mor phol ogy of dy ing cells us ing
Hochest33258, flourescein diacetate and propidium
io dide stain ing. The ir ra di ated HIEC cells pretreated
with hy dro gen-rich PBS dem on strated a pro tec tive ef -
fect with the re duced num ber of apoptotic cells to
26.1% as com pared to 49.3% in PBS-pretreated ir ra di -
ated cells, fig. 2(c) and (d). These data sug gest that H2
can at ten u ate apoptosis in ir ra di ated HIEC cells.
Treating AHH-1 cells with hydrogen be fore
ir ra di a tion could in crease endogenous
an ti ox i dant sta tus
The lev els of en zy matic an ti ox i dants (SOD) and
the ac tiv i ties of non-en zy matic an ti ox i dant (GSH) are
shown in fig. 3. Gamma ir ra di ated lym pho cytes showed
a sig nif i cant de crease in the lev els of both en zy matic
and non-en zy matic an ti ox i dant sta tus when com pared
to hy dro gen-rich PBS pretreated groups. The re sults in -
di cated that pre treat ment with H2 could re store the an ti -
ox i dant sta tus.
L. Qian, et al.: Hy dro gen-Rich PBS Pro tects Cul tured Hu man Cells from ...
Nu clear Tech nol ogy & Ra di a tion Pro tec tion: Year 2010, Vol. 25, No. 1, pp. 23-29 25
Fig ure 1. The treated HIEC cells were ir ra di ated with 0, 1,
2, 4, and 8 Gy and plated for clonogenic sur vival as say and
for LDH leak age as say; the sur viv ing frac tions (a) and
changes in the lev els of LDH in nor mal, g-ir ra di ated and
H2 pretreated lym pho cytes from three ex per i ments (b) are
shown; values are given as mean ±S. E. M. *P < 0.05
Hydrogen-rich PBS pro tects lipids and
nu clear DNA of AHH-1cells from
peroxidation in duced by ra di a tion
As shown in fig. 4(a), cel lu lar MDA con cen tra -
tion at 4 hours af ter the ir ra di a tion in the H2 group was
sig nif i cantly lower than that of the con trol group. This
re sult in di cated that H2 could pro tect lipids from
peroxidation in duced by ra di a tion. As shown in fig.
4(b), H2 com pa ra bly de creased the con cen tra tion of
8-OHdG rel a tive to the con trol group, in di cat ing that
H2 can pro tect DNA from peroxidation in duced by ra -
di a tion.
DIS CUS SION
This study shows that hy dro gen can sig nif i -
cantly pro tect hu man cells from ion iz ing ra di a tion. In -
ha la tion of H2 was re ported to pro tect ce re bral [1],
myo car dial [2], and hepatic [9] I/R in jury in an i mal
mod els in sev eral re cent stud ies. Also, it is re ported
that hy dro gen in ha la tion ame lio rates ox i da tive stress
in trans plan ta tion in duced in tes ti nal graft in jury [3].
Since most of the ion iz ing ra di a tion-in duced cel lu -
lar dam age is caused by hydroxyl rad i cals, we spec u late
that the radioprotective ef fect may re sult from its rad i cal
ox y gen spe cies (ROS) scav eng ing ef fect. It was re ported
L. Qian, et al.: Hy dro gen-Rich PBS Pro tects Cul tured Hu man Cells from ...
26 Nu clear Tech nol ogy & Ra di a tion Pro tec tion: Year 2010, Vol. 25, No. 1, pp. 23-29
Fig ure 2. Hy dro gen-rich PBS at tenu ates ra di a tion-in duced apoptosis in HIEC cells. The treated cells were col lected 24
hours af ter the ir ra di a tion, stained with Annexin V-APC and propidium io dide and an a lyzed by flow cytometry. The rep -
re sen ta tive di a grams of dis tri bu tion of the stained cells (a) and a bar graph of apoptotic cells ex pressed as a per cent of to tal
cells are shown. Val ues are given as mean ±S. E. M. (n = 4); *P < 0.01 (b); cells were stained with FDA, Hoechst33258 and
PI 24 hours af ter ir ra di a tion and apoptotic cells were counted in mul ti ple ran domly se lected fields. The rep re sen ta tive mi -
cro graphs (c) and a bar graph of apoptotic cells ex pressed as a per cent of to tal cells are shown; values are given as mean
±S. E. M. (n = 4); *P < 0.01 (d)
that the ef fect of free rad i cal scav en gers could ame lio rate
the ox i da tive in ju ries due to ion iz ing ra di a tion [10, 11].
The sulfhydryl com pound amifostine (WR-2721),
which is the only radioprotectant reg is tered in use for hu -
mans, has shown good radioprotective ef fects [12].
How ever, when it was ad min is tered by in jec tion, it
caused many neg a tive ef fects such as vom it ing, hy per -
ten sion, nau sea, and other side ef fects caused by the tox -
ic ity [13, 14]. Some other radioprotectors, such as nat u ral
an ti ox i dants, vi ta min E, flavonoids and oth ers, have
fewer toxic side ef fects but also a lower de gree of pro tec -
tion com pared to thiol agents [13], and cytokines and
immunomodulators should be used with low ra di a tion
doses or in com bi na tion with rad i cal scav en gers and an ti -
ox i dants [15]. How ever, it is phys i o log i cally safe for hu -
mans to in hale hy dro gen at a rel a tively low con cen tra -
tion, be cause hy dro gen is con tin u ously pro duced by
co lonic bac te ria in the body and nor mally cir cu lates in
the blood [16]. It is a highly diffusible gas which could
elim i nate hydroxyl rad i cal [17]. Dis solv ing H2 in sol -
vents such as PBS makes this ex plo sive gas more safe for
clin i cal use.
Rad i cal ox y gen spe cies O2– and H2O2 are de tox -
i fied by an ti ox i dant de fense en zymes, un like ·OH and
ONOO–, which so far could not be detoxified by any
an ti ox i dant de fense en zyme. Hy dro gen gas se lec -
tively re duces these two det ri men tal ROS [1]. A
hydroxyl rad i cal is the most re ac tive prod uct of re ac -
tive ox y gen spe cies gen er ated in cells. Cellular
macromolecules, such as DNA, pro teins, and lipids,
can eas ily re act with hydroxyl rad i cals to ex ert a
cytotoxic ef fect.
An ti ox i dant en zymes (SOD) are im por tant in
pro vid ing pro tec tion from ra di a tion ex po sure [18] and
glutathione (GSH) par tic i pates non-en zy mat i cally in
pro tec tion against ra di a tion dam age [19]. En dog e nous
an ti ox i dants are a group of sub stances which could
sig nif i cantly in hibit or de lay ox i da tive pro cesses [20].
A num ber of harms can re sult from a re duc tion of the
ac tiv ity of these sub stances. DNA is one of the ma jor
L. Qian, et al.: Hy dro gen-Rich PBS Pro tects Cul tured Hu man Cells from ...
Nu clear Tech nol ogy & Ra di a tion Pro tec tion: Year 2010, Vol. 25, No. 1, pp. 23-29 27
Fig ure 3. Changes in the ac tiv i ties of SOD and GSH in nor mal, g-ir ra di ated, and H2 pretreated lym pho cytes; values are
given as mean ±S. E. M. (n = 4); *P < 0.01.
Fig ure 4. Hy dro gen-rich PBS sig nif i cantly de creased the lev els of MDA, a marker of ox i da tive stress (a), and ox i da tive
DNA dam age as sessed by 8-OHdG immunoreactivity. 8-OHdG con cen tra tion in nor mal, g-ir ra di ated, and H2 pretreated
groups (b) half an hour af ter the ir ra di a tion are shown; rel a tive to the con trol group, H2 sig nif i cantly de creased the con -
cen tra tion of 8-OHdG; val ues are mean ±S. E. M. (n = 6); *P < 0.01.
tar gets of free rad i cals, and 8-OHdG is formed from
deoxyguanosine in DNA by hydroxyl free rad i cals
[21]. Also, mem brane lipids are the ma jor tar gets of
free rad i cals [22]. The in crease in the lev els of lipid
peroxidation prod ucts such as malondialdehyde and
TBARs is the in di ca tion of mem brane lipid dam age
[23]. In our study, we found that the pretreatment of
hy dro gen-rich PBS prior to ra di a tion ex po sure in -
creased the an ti ox i dant sta tus at both en zy mic and
non-en zy mic lev els and de creased the levels of MDA
and 8-OHdG com pared with the cells pretreated with -
out hy dro gen-rich PBS. We may con clude that the in -
creases of the an ti ox i dant sta tus have fur ther de -
creased the at tack of free rad i cals on biomolecules
in clud ing DNA and mem brane lipids and thereby de -
creased the del e te ri ous ef fects of ra di a tion on cells.
In con clu sion, hy dro gen-rich PBS could pro tect
hu man cells from ra di a tion. This radioprotective ef -
fect may re sult from its rad i cal ox y gen spe cies scav -
eng ing ef fect. Dis solv ing hy dro gen in so lu tion (PBS,
wa ter, sa line) makes it safer and more con ve nient to
use in clinic. We be lieve that hy dro gen gas, es pe cially
hy dro gen-rich so lu tion, may give us more hope for
greater pro tec tion from ir ra di a tion.
AC KNOWL EDGE MENTS
This work was sup ported by a grant from the Na -
tional Nat u ral Sci ence Foun da tion of China (No.
30770503).
REF ER ENCES
[1] Ohsawa, I., Ishikawa, M., Takahashi, K., Watanabe,
M., Nishimaki, K., Yamagata, K., Katsura, K.,
Katayama, Y., Asoh, S., Ohta, S., Hy dro gen Acts as a
Ther a peu tic An ti ox i dant by Se lec tively Re duc ing
Cytotoxic Ox y gen Rad i cals, Nat Med., 13 (2007), pp.
688-694
[2] Hayashida, K., Sano, M., Ohsawa, I., Shinmura, K.,
Tamaki, K., Kimura, K., Endo, J., Katayama, T.,
Kawamura, A., Kohsaka, S., Makino, S., Ohta, S.,
Ogawa, S., Fukuda, K., In ha la tion of Hy dro gen Gas
Re duces In farct Size in the Rat Model of Myo car dial
Ischemia-Reperfusion In jury, Biochem Biophys Res
Commun, 373 (2008), 1, pp. 30-35
[3] Buchholz, B. M., Kaczorowski, D. J., Sugimoto, R.,
Yang, R., Wang, Y., Billiar, T. R., McCurry, K. R.,
Bauer, A. J., Nakao, A., Hy dro gen In ha la tion Ame lio -
rates Ox i da tive Stress in Trans plan ta tion In duced In -
tes ti nal Graft In jury, Am J. Trans plant, 8 (2008), 10,
pp. 2015-2024
[4] Abraini, J. H., Gardette-Chauffour, M. C., Mar ti nez,
E., Rostain, J. C., Lemaire, C., Psychophysiological
Re ac tions in Hu mans dur ing an Open Sea Dive to
500 m with a Hy dro gen-He lium-Ox y gen Mix ture, J.
Appl Physiol, 76 (1994), 3, pp. 1113-1118
[5] Ward, J. F., DNA Dam age Pro duced by Ion iz ing Ra di -
a tion in Mam ma lian Cells, Iden ti ties, Mech a nisms of
For ma tion, and Reparability, Prog Nu cleic Acid Res
Mol Biol., 35 (1988), pp. 95-125
[6] Agrawal, A., Kale, R. K., Ra di a tion In duced
Peroxidative Dam age, Mech a nism and Sig nif i cance,
In dian J. Exp Biol., 39 (2001), 4, pp. 291-309
[7] Prasad, N. R., Menon, V. P., Vasudev, V., Pugalendi,
K. V., Radioprotective Ef fect of Sesamol on
Gamma-Ra di a tion In duced DNA Dam age, Lipid
Peroxidation and An ti ox i dants Lev els in Cul tured
Hu man Lym pho cytes, Tox i col ogy, 209 (2005), 3, pp.
225-235
[8] Yazlovitskaya, E. M., Ed wards, E., Thotala, D., Fu,
A., Osusky, L. K., Whetsell, Jr., W. O., Boone, B.,
Shinohara, E. T., Hallahan, D. E., Lith ium Treat ment
Pre vents Neurocognitive Def i cit Re sult ing from Cra -
nial Ir ra di a tion, Can cer Res, 66 (2006), 23, pp.
11179-11186
[9] Fukuda, K., Asoh, S., Ishikawa, M., Yamamoto, Y.,
Ohsawa, I., Ohta, S., In ha la tion of Hy dro gen Gas Sup -
presses Hepatic In jury Caused by Ischemia/Reperfusion
through Re duc ing Ox i da tive Stress., Biochem Biophys
Res Commun, 361 (2007), 3, pp. 670-674
[10] Shirazi, A., Ghobadi, G., Ghazi-Khansari, M.. A
Radiobiological Re view on Melatonin, A Novel
Radioprotector (To kyo), J. Radiat Res, 48 (2007), 4,
pp. 263-272
[11] Yan, S. X., Hong, X. Y., Hu, Y., Liao, K. H., Tempol,
One of Nitroxides, is a Novel Ul tra vi o let-A1 Ra di a -
tion Pro tec tor for Hu man Der mal Fibroblasts, J.
Dermatol Sci., 37 (2005), 3, pp. 137-143
[12] Gosselin, T. K., Mautner, B., Amifostine as a
Radioprotectant, Clin J. Oncol Nurs, 6 (2002), 3, pp.
175-176
[13] Hosseinimehr, S. J., Trends in the De vel op ment of
Radioprotective Agents, Drug Discov To day, 12
(2007), 19-20, pp. 794-805
[14] Genvresse, I., Lange, C., Schanz, J., Schweigert, M.,
Harder, H., Possinger, K., Späth-Schwalbe, E.,
Tolerability of the Cytoprotective Agent Amifostine
in El derly Pa tients Re ceiv ing Che mo ther apy: a Com -
par a tive Study, Anticancer Drugs, 12 (2001), 4, pp.
345-349
[15] Herodin, F., Drouet, M., Cytokine-Based Treat ment
of Ac ci den tally Ir ra di ated Vic tims and New Ap -
proaches, Exp Hematol, 33 (2005), 10, pp. 1071-1080
[16] Reth, M., Hy dro gen Per ox ide as Sec ond Mes sen ger in
Lym pho cyte Ac ti va tion, Nat Immunol, 3 (2002), 12,
pp. 1129-1134
[17] Labiche, L. A., Grotta, J. C., Clin i cal Tri als for
Cytoprotection in Stroke, Neuro Rx, 1 (2004), 1, pp.
46-70
[18] el-Nahas, S. M., Mattar, F. E., Mohamed, A. A.,
Radioprotective Ef fect of Vi ta mins C and E, Mutat
Res., 301 (1993), 2, pp. 143-147
[19] Kojima, S., Shimomura, H., Matsumori, S., Ef fect of
Pre-Ir ra di a tion with Low-Dose Gamma-Rays on
Chem i cally In duced Hepatotoxicity and Glutathione
De ple tion, Anticancer Res, 20 (2000), 3A, pp.
1583-1588
[20] Kalpana, K. B., Devipriya, N., Srinivasan, M.,
Menon, V. P., In ves ti ga tion of the Radioprotective Ef -
fi cacy of Hesperidin Against Gamma-Ra di a tion In -
duced Cel lu lar Dam age in Cul tured Hu man Pe riph -
eral Blood Lym pho cytes, Mutat Res, 676 (2009), 1-2,
pp. 54-61
[21] Kasai, H., Anal y sis of a Form of Ox i da tive DNA
Dam age, 8-hydroxy-2’-deoxyguanosine, as a Marker
of Cel lu lar Ox i da tive Stress Dur ing Carcinogenesis,
Mutat Res, 387 (1997), 3, pp. 147-163
[22] Serbinova, E., Kagan, V., Han, D., Packer, L., Free
Rad i cal Re cy cling and Intramembrane Mo bil ity in
the An ti ox i dant Prop er ties of Al pha-Tocopherol and
L. Qian, et al.: Hy dro gen-Rich PBS Pro tects Cul tured Hu man Cells from ...
28 Nu clear Tech nol ogy & Ra di a tion Pro tec tion: Year 2010, Vol. 25, No. 1, pp. 23-29
Al pha-Tocotrienol, Free Radic Biol Med, 10 (1991),
pp. 263-275
[23] Dubner, D., Gisone, P., Jaitovich, I., Perez, M., Free
Rad i cals Pro duc tion and Es ti ma tion of Ox i da tive
Stress Re lated to Gamma Ir ra di a tion, Biol Trace Elem
Res, 47 (1995), pp. 265-270
Re ceived on Sep tem ber 3, 2009
Ac cepted on Feb ru ary 8, 2010
L. Qian, et al.: Hy dro gen-Rich PBS Pro tects Cul tured Hu man Cells from ...
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Liren ]EN, Bailung LI, Fei CAO, @u~eng HUANG,
[ulin LIU, \enming CAI, Fu GAO
PBS OBOGA]EN VODONIKOM [TITI ^OVE^IJE ]ELIJE U
KULTURI OD ZRA^EWEM NANETIH OZLEDA
Vodoni~ni radikali imaju zna~ajnu ulogu u indukciji }elijskih ozleda dejstvom
jonizuju}eg zra~ewa. Molekularni vodonik mo`e selektivno da smawi koncentraciju
hidroksilnih radikala in vi tro. Takve osobine ~ine ga potencijalnim radioprotektorom. U ovom
radu testirana su radioprotektivna svojstva vodonika in vi tro kori{}ewem PBS oboga}enog
vodonikom. Tretirawe }elija s PBS oboga}enim vodonikom, pre ozra~ivawa, zna~ajno smawuje
apoptozu indukovanu jonizuju}im zra~ewem, pove}ava vitalnost i pre`ivqavawe ozra~enih
kripti~nih }elija tankog creva (HIEC), pove}ava koncentraciju endogenih antioksidanata i
smawuje koncentraciju 8-hidroksideoksiguanozin-malondialdehida u limfocitima (AHH-1
}elije). Iz dobijenih rezultata mo`e se zakqu~iti da vodonik poseduje zna~ajnu osobinu
radioprotektora: efikasan je i nema ne`eqenih svojstava.
Kqu~ne re~i: vodonik, jonizuju}e zra~ewe, za{tita od zra~ewa, apoptoza, reaktivne
jjjjjjjjjjjjjjj.jjjjjjjjkiseoni~ne vrste
