Comparison of micronuclei frequency in bone marrow cells of three rat lines
The aim of this paper is to compare the spontaneous and induced with cyclophosphamide micronucleus indexes in bone marrow cells of the Sprague Dawley, Lewis and Wistar rat lines. Five experimental groups were formed (10 animals of each sex and of each line, in every group). The first group was used...
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Інститут клітинної біології та генетичної інженерії НАН України
2013
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| Цитувати: | Comparison of micronuclei frequency in bone marrow cells of three rat lines / D.F. Arencibia-Arrebola, L.A. Rosario-Fernandez, Y.E. Suarez-Fernandez, A. Vidal-Novoa // Цитология и генетика. — 2013. — Т. 47, № 2. — С. 70-76. — Бібліогр.: 26 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1859980909405536256 |
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| author | Arencibia-Arrebola, D.F. Rosario-Fernandez, L.A. Suarez-Fernandez, Y.E. Vidal-Novoa, A. |
| author_facet | Arencibia-Arrebola, D.F. Rosario-Fernandez, L.A. Suarez-Fernandez, Y.E. Vidal-Novoa, A. |
| citation_txt | Comparison of micronuclei frequency in bone marrow cells of three rat lines / D.F. Arencibia-Arrebola, L.A. Rosario-Fernandez, Y.E. Suarez-Fernandez, A. Vidal-Novoa // Цитология и генетика. — 2013. — Т. 47, № 2. — С. 70-76. — Бібліогр.: 26 назв. — англ. |
| collection | DSpace DC |
| container_title | Цитология и генетика |
| description | The aim of this paper is to compare the spontaneous and induced with cyclophosphamide micronucleus indexes in bone marrow cells of the Sprague Dawley, Lewis and Wistar rat lines. Five experimental groups were formed (10 animals of each sex and of each line, in every group). The first group was used as the negative control (intact animals), the second one was exposed to oral administration of drugs; other conditions were the same as for the other groups. The third group was treated with 2 % Tween 65 and the fourth group was treated with 0.9 % NaCl. Both substances were administered by oral way to 2 ml/kg during 14 days. The fifth group was treated intraperitoneally with strong mutagen cyclophosphamide in the dose of 50 mg/kg (10 ml/kg in solution), on 48th and 24th hours before euthanasia. The Sprague Dawley line (both sexes) was significantly different from the other lines. Rats of this line had lower index of spontaneous formation of micronuclei, higher index of cyclophosphamideinduced micronucle formation, percent of micronucleated erythrocytes in bone marrow and the index of cytotoxicity. The results obtained make it possible to identify the most appropriate line of rats as model animals for studies of genotoxicity. It will allow also to obtain more accurate estimates of genotoxicity of various substances.
Сравнивали спонтанные и индуцированные циклофосфамидом микроядерные индексы в клетках костного мозга трех линий крыс – Sprague Dawley, Льюис и Вистар. Сформировали пять экспериментальных групп (по 10 животных каждого пола и линии в каждой группе). Первую группу использовали в качестве негативного контроля (интактные животные), вторая служила контролем влияния перорального приема препаратов. Все условия содержания были одинаковыми для всех групп животных. Крыс третьей группы обрабатывали 2 % Tween 65, а четвертой группы – 0,9 % NaCl. Оба вещества вводили перорально в дозе 2 мл/кг в течение 14 дней. Животных пятой группы обрабатывали внутрибрюшинно циклофосфамидом в дозе 50 мг/кг (10 мл/кг раствора), за 48 и 24 ч до эвтаназии. Линия Sprague Dawley (оба пола) существенно отличается от других. Крысы этой линии имели низкие показатели спонтанного образования микроядер и высокий индекс при индукции циклофосфамидом, а также процент эритроцитов с микроядрами в костном мозге и индекс цитотоксичности. Полученные результаты позволяют определить наиболее подходящие линии крыс в качестве модельных животных для изучения генотоксичности. Это также позволит получать более точные оценки генотоксичности различных веществ. Порівнювали спонтанні та індуковані циклофосфамідом мікроядерні індекси в клітинах кісткового мозку трьох ліній щурів – Sprague Dawley, Люіс і Вистар.
Сформували п’ять експериментальних груп (по 10 тварин кожної статі і лінії в кожній групі). Першу групу використали як негативний контроль (інтактні тварини), друга слугувала контролем впливу перорального прийому препаратів. Всі умови утримання були однаковими для всіх груп тварин. Щурів третьої групи обробляли 2 % Тween 65, а четвертої групи – 0,9 % NaCl. Обидві речовини вводили перорально в дозі 2 мл/кг впродовж 14 днів. Тварин п’ятої групи обробляли внутрішньочеревно циклофосфамідом в дозі 50 мг/кг (10 мл/кг розчину), за 48 і 24 год до евтаназії. Лінія Sprague Dawley (обидві статі) істотно відрізнялась від інших. Щури цієї лінії мали низькі показники спонтанного утворення мікроядер і високий індекс при індукції циклофосфамідом, а також відсоток еритроцитів з мікроядрами в кістковому мозку та індекс цитотоксичності. Отримані результати дозволяють визначити найбільш відповідні лінії щурів як модельних тварин для вивчення генотоксичності. Це також дозволить отримувати точніші оцінки генотоксичності різних речовин.
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| first_indexed | 2025-12-07T16:26:18Z |
| format | Article |
| fulltext |
70 ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2013. ¹ 2
D.F. ARENCIBIA-ARREBOLA 1,
L.A. ROSARIO-FERNANDEZ 2, Y.E. SUAREZ-FERNANDEZ 3,
A. VIDAL-NOVOA 4
1 Finlay Institute, 17 Avenue, Playa Municipality, Havana, Cuba
E-mail: darencibia@finlay.edu.cu
2 Institute of Pharmacy and Food Science, Havana, Cuba
3 Medicine Veterinary Faculty UNAH, Cuba
4 Faculty of Biology, Havana University, Cuba
COMPARISON OF MICRONUCLEI
FREQUENCY IN BONE MARROW CELLS
OF THREE RAT LINES
The aim of this paper is to compare the spontane-
ous and induced with cyclophosphamide micronucleus
indexes in bone marrow cells of the Sprague Dawley,
Lewis and Wistar rat lines. Five experimental groups
were formed (10 animals of each sex and of each line,
in every group). The first group was used as the nega-
tive control (intact animals), the second one was exposed
to oral administration of drugs; other conditions were
the same as for the other groups. The third group was
treated with 2 % Tween 65 and the fourth group was
treated with 0.9 % NaCl. Both substances were admin-
istered by oral way to 2 ml/kg during 14 days. The fifth
group was treated intraperitoneally with strong mutagen
cyclophosphamide in the dose of 50 mg/kg (10 ml/kg in
solution), on 48th and 24th hours before euthanasia. The
Sprague Dawley line (both sexes) was significantly dif-
ferent from the other lines. Rats of this line had lower
index of spontaneous formation of micronuclei, higher in-
dex of cyclophosphamide-induced micronucle formation,
percent of micronucleated erythrocytes in bone marrow
and the index of cytotoxicity. The results obtained make
it possible to identify the most appropriate line of rats as
model animals for studies of genotoxicity. It will allow
also to obtain more accurate estimates of genotoxicity of
various substances.
Introduction. Usually for testing genotoxicity of
different substances researchers use different lines
of rats. Due to genetic differences between lines
their results cannot be compared. That is why the
correct choosing of model organisms remains the
very important problem [1]. Comparative studies
of spontaneous and induced genome damages in
different lines of rats could provide researchers
with efficient way of choosing the most appropri-
ate genetic lines of rats for their tests.
One of popular test of genotoxicity is the
micronucleus assay in bone marrow cells. This
test is easy to perform and it provides research-
ers with clear information on the rate of genome
damages in bone marrow cells. This approach al-
lows to register in vivo an ability of chemical sub-
stances to induce chromosome breaks [2]. The
increasing of frequency of polychromatic eryth-
rocytes with micronuclei indicates the respective
invreasing of chromosome damages [2, 3]. The
detection of micronucleated erythrocytes can
also be performed in species, whose spleen can-
not eliminate the micronucleated erythrocytes or
species sufficiently sensitive to detect agents that
cause numeric or structural chromosome aberra-
tions [2, 3]. The micronucleus assay must include
the positive control with mutagenic substance –
usually cyclophosphamide [2]. This mutagen is
an alquilant agent that forms monoadducts and
cross-connections between chains [3] and does
not possess the specificity dependent on the cell
cycle [4]. The aim of this article is to compare
the spontaneous and cyclophosphamide induced
micronucleus indexes of Sprague Dawley, Lewis
and Wistar rat lines.
Materials and methods. Animals and environ-
mental conditions. Sprague Dawley, Lewis and
Wistar rat lines, young adults (6–8 weeks of age)
of both sexes with body mass about 180–210 g at
the end of the quarantine were used in our work.
They were held under temperature 23 ± 2 °C,
relative humidity 60 ± 10 % and 12 h cycles of
light-darkness. The access to the water and the
food (CENPALAB) was ad libitum. These condi-
tions were common for all groups. We adhered
to the ethical principles for the investigation with
laboratory animals according to the Canadian
Advice of Care Animal [5].
Doses and treatment procedures. All the sub-
stances were administered in the «two days» shed-
ule and the concentrations were adjusted weekly
according to the body mass increasing. The ani-© D.F. ARENCIBIA-ARREBOLA, L.A. ROSARIO-FERNANDEZ,
Y.E. SUAREZ-FERNANDEZ, A.VIDAL-NOVOA, 2013
71
Comparison of micronuclei frequency in bone marrow cells of three rat lines
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2013. ¹ 2
mals were randomly distributed among 5 groups
with 10 rats of each sex of each line per group.
The first group was used as the negative control
(intact animals), the second one was exposed to
oral administration of drugs (technique control
group). The third experimental group was treated
with 2 % Tween 65 (carrying substance 1) which
is used as carrier of many nonpolar hydrophobic
substances, useful as tensoactive agent [6]; the
fourth group was treated with 0.9 % NaCl (carry-
ing sustance 2), useful as solvent of most hydro-
philic substances [7]. Both substances were pre-
pared for 2 hours before using and were applied
per os to 2 ml/kg during a period of 14 days.
The fifth group was treated intraperitoneally with
cyclophosphamide (Ledoxina®. Lemery. CORP)
previously dissolved in 0,9 % NaCl, in the dose
of 50 mg/kg [8]. The solution of cyclophospha-
mide was used immediately after being prepared
and applied to animals (in dose 10 ml/kg), on the
48th and 24th hours before euthanasia [8].
Clinical observations. We performed two daily
clinical observations, during 8:30–10:30 a.m. and
during 3:00–4:30 p.m. During each observation
we had investigated the general clinical state of
the animal, including the palpation for the detec-
tion of lesions, possible breathing affectations, of
the nervous, cardiovascular, gastrointestinal sys-
tem, state of the skin, hair, mucous and eyes col-
oration.
Euthanasia method. All animals were sacrificed
with ether. We provided this process in a way that
the animals did not show pain signs or suffering.
Bone marrow cells micronucleus assay. The mi-
cronucleus assay was performed according to the
modified protocol [9, 10]. One femur per animal
was extracted and the medullar cavity was washed
out with 3 mL of fetal bovine serum. The ob-
tained cells were centrifuged at 200 g for 10 min;
and the pellet was spreaded onto the slides [9].
The slides (minimum 2 slides per animal) were
dried 24 h under ambient temperature, fixed in
absolute methanol (5 min) and stained with 5 %
Giemsa for 12–15 min [10]. The analysis was
performed by three independent observers, us-
ing a microscope Olympus BH-2 (100× lens with
immersion).
The polychromatic erythrocytes (PE) and
normochromatic erythrocytes (NE) were count-
ed among 2000 cells/animal. The young eryth-
rocytes are the PE, microscopically they are ob-
served as blue cells and the normochromatic
erythrocytes (adults) are observed as pink [2, 9].
The percent of PE with micronucleus (MN-EP)
was calculated in 2000 PE/animal. Later the cy-
totoxicity index (PE/NE) was calculated for the
total population of erythrocytes per group and
total number of micronucleus (MN) [10]. In this
work we presented the average results of three
independent observers.
Statistical analysis. We calculated the aver-
age (X) and standard deviation (S.D) of studed
variables. The comparisons between controls and
cyclophosphamide-treated (CF) groups for the
same line, among different lines, sex and ob-
servers were performed with ANOVA with sig-
nificance level = 0.05. The categorical variables
(total number of micronuclei) were analysed by
Chi-Square, with significance level of = 0.01.
All the analyses were performed with STATIS-
TICA (version 6) (StatSoft, Inc, 2003).
Permission of ethics committee. The authors
declare that this work was made on the base of
good practices of preclinical laboratory that are
present in the national regulation of protocols of
research, approved in the Cuban republic. When
this research began it was obtained the protocol
approval in writing.
Results and discussion. We did nît observe any
systemic damages of the whole organism of animals.
We did nît observe also animals with symptoms of
intoxication. The damages were noted at the level
of somatic cells. It let us conclude that the doses
and the schedule of treatments were appropriate.
The groups of ñyclophosphamide-treated ani-
mals differed from other groups of rats, while
groups of negative controle, groups of control
of the treatment way and two solvent substances
did not differe each from other in the same sex
groups and lines [7, 11]. The results obtained for
Sprague Dawley rats are in good accordance with
our previous works [12, 13].
The low (but significant) intoxication (PE/
NE) was noted for Sprague Dawley rats of both
sexes. No differences were found among sexes of
the same group and rat line (Table 1) [7, 11]. In
other studies differences among sexes were not
observed also [14–16].
In Sprague Dawley line the PE/NE is equal
1.18 ± 0.06; the genotoxicity index was about
Table 1
The cytotoxicity indexes (PE/NE) and percentage of PE with micronucleus in bone marrow
of Sprague Dawley, Lewis and Wistar rats
Groups n Sex MNR PE/NET MN–PE (%)T
Sprague Dawley Rats
Negative Control (Intact)
Technique Control
Vehicle Substance 1
Vehicle Substance 2
Cyclophosphamide 1
10
10
10
10
10
10
10
10
10
10
F
M
F
M
F
M
F
M
F
M
28a
26a
32a
27ac
29a
30a
25a
22a
250**a
241**ac
1,21 ± 0,05
1,17 ± 0,04
1,19 ± 0,06a
1,20 ± 0,03a
1,21 ± 0,02a
1,19 ± 0,04a
1,18 ± 0,04a
1,18 ± 0,06a
0,89 ± 0,04*a
0,90 ± 0,03*a
0,20 ± 0,03a
0,18 ± 0,04a
0,22 ± 0,01a
0,19 ± 0,05a
0,20 ± 0,02a
0,21 ± 0,02a
0,17 ± 0,05a
0,15 ± 0,06a
1,74 ± 1,03*a
1,68 ± 0,92*ac
Lewis Rats
Negative Control (Intact)
Technique Control
Vehicle Substance 1
Vehicle Substance 2
Cyclophosphamide1
10
10
10
10
10
10
10
10
10
10
F
M
F
M
F
M
F
M
F
M
46b
48b
48b
47b
49b
50b
42 b
44 b
295**b
291**b
1,10 ± 0,03b
1,11 ± 0,03b
1,11 ± 0,02b
1,09 ± 0,03b
1,12 ± 0,02b
1,13 ± 0,02 b
1,10 ± 0,03 b
1,13 ± 0,02 b
0,82 ± 0,02*b
0,81 ± 0,02*b
0,31 ± 0,04b
0,33 ± 0,03b
0,33 ± 0,05b
0,32 ± 0,06b
0,34 ± 0,06b
0,34 ± 0,05b
0,29 ± 0,07b
0,30 ± 0,07b
2,03 ± 1,48*b
2,00 ± 1,08*b
Wistar Rats
Negative Control (Intact)
Technique Control
Vehicle Substance 1
Vehicle Substance 2
Cyclophosphamide1
10
10
10
10
10
10
10
10
10
10
F
M
F
M
F
M
F
M
F
M
53b
47bc
54b
49bc
50b
55bc
51b
46bc
292**bc
298**b
1,11 ± 0,02b
1,10 ± 0,04b
1,12 ± 0,03b
1,14 ± 0,02b
1,10 ± 0,03b
1,11 ± 0,03b
1,10 ± 0,03b
1,12 ± 0,04b
0,79 ± 0,03*b
0,80 ± 0,02*b
0,36 ± 0,03b
0,33 ± 0,02b
0,37 ± 0,02b
0,34 ± 0,04b
0,34 ± 0,05b
0,38 ± 0,03b
0,35 ± 0,03b
0,32 ± 0,05b
2,01 ± 1,56*b
2,05 ± 1,16*b
Inducation. PE – Polychromatic Erythrocytes. NE – Normochromatic Erythrocytes. MN – Micronucleus.
Count in 2000 cells/animal of 10 animal/group/sex/line. T Analyzed variable using the ANOVA test (X: average;
SD: Standard Deviation). R Analysed variable, 2 non parametric test. 1 Intraperiytoneal treatment. *p < 0,05
(comparison with the negative control group in the same genetic line and sex, ANOVA test). **p < 0,01
(comparison with the negative control group in the same genetic line and sex, Chi Square ( 2) non parametric
test). a, b – Different letters it differs when comparing among lines keeping in mind the same variable in the
same experimental group and sex. c – Comparison between sexes in the same experimental group and rat lines.
D.F. Arencibia-Arrebola, L.A. Rosario-Fernandez, Y.E. Suarez-Fernandez, A.Vidal-Novoa
72 ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2013. ¹ 2
73ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2013. ¹ 2
Comparison of micronuclei frequency in bone marrow cells of three rat lines
Table 2
Variation of data obtained by three observers
Groups Sex
MNR PE/NET MN–PE (%)T
Ob1 Ob2 Ob3 Ob1 Ob2 Ob3 Ob1 Ob2 Ob3
Sprague Dawley Rats
NC
TC
VS1
VS2
CF
F
M
F
M
F
M
F
M
F
M
25a
24a
27a
22a
26a
33a
20a
20a
241a
235a
30a
24a
34a
29a
27a
30a
29b
19a
249ab
248a
29a
30a
35b
30b
34b
27a
26ab
27b
260b
240a
1,26b
1,14a
1,18a
1,22a
1,26b
1,16a
1,17a
1,15a
0,79a
0,94a
1,18a
1,19a
1,23a
1,20a
1,19a
1,20a
1,18a
1,20a
0,98a
0,77a
1,19a
1,18a
1,16a
1,19a
1,18a
1,21a
1,19a
1,19a
0,90a
0,99a
0,15a
0,22a
0,25a
0,17a
0,16a
0,22a
0,16a
0,15a
1,82b
1,55a
0,19a
0,15b
0,22a
0,19a
0,20a
0,24a
0,15a
0,13a
1,56a
1,62a
0,26a
0,17a
0,19a
0,21a
0,24a
0,17a
0,20a
0,17a
1,84a
1,87a
Lewis Rats
NC
TC
VS1
VS2
CF
F
M
F
M
F
M
F
M
F
M
50a
53a
43a
43a
43a
42a
40a
47a
286a
280a
43a
46a
46a
45a
55b
52ab
46a
45a
289ab
288ab
45a
45a
55a
53a
49ab
56b
40a
40a
310b
305b
1,10a
1,17b
1,08a
1,15a
1,09a
1,08a
1,12a
1,15a
0,66a
0,90b
1,13a
1,10ab
1,11a
1,07a
1,10a
1,16a
1,11a
1,14a
0,92b
0,83ab
1,07a
1,06a
1,14a
1,06a
1,17a
1,15a
1,07a
1,10a
0,88ab
0,70a
0,29ab
0,36a
0,38a
0,29a
0,34a
0,25a
0,34a
0,30a
2,09ab
2,50b
0,27a
0,34a
0,28a
0,30a
0,32a
0,38ab
0,26a
0,33a
1,76a
1,84a
0,37b
0,29a
0,34a
0,37a
0,36a
0,39b
0,27a
0,27a
2,25b
1,67a
Wistar Rats
NC
TC
VS1
VS2
CF
F
M
F
M
F
M
F
M
F
M
42ab
42a
55a
46a
44a
51a
49a
44a
285a
299a
50b
49a
48a
49a
56b
58a
51a
45a
296a
300a
37a
50a
59a
52a
50ab
56a
53a
49a
295a
295a
1,10a
1,07a
1,13a
1,14a
1,11a
1,10a
1,07a
1,13a
0,87b
0,83a
1,13a
1,14a
1,09a
1,17a
1,12a
1,15a
1,12a
1,16a
0,90b
0,78a
1,10a
1,09a
1,14a
1,11a
1,07a
1,08a
1,11a
1,07a
0,60a
0,80a
0,35a
0,29a
0,39a
0,34a
0,33a
0,37a
0,32a
0,33a
1,94a
2,09a
0,35a
0,37a
0,37a
0,37a
0,35a
0,36a
0,36a
0,32a
2,04a
2,00a
0,39a
0,33a
0,35a
0,32a
0,34a
0,42a
0,37a
0,31a
2,05a
2,06a
Inducation. NC – Negative Control. TC – Technique Control. VS1 – Vehicle Substance 1. VS2 – Vehicle
Substance 2. CF – Cyclophosphamide. O – number of observers. PE – Polychromatic Erythrocytes. NE –
Normochromatic Erythrocytes. MN – Micronucleus. Count in 2000 cells/animal of 10 animal/group/sex/line.
T Analyzed variable using the ANOVA test (X: average; SD: Standard Deviation, p < 0,05). RAnalyzed variable
using the 2 non parametric test (p < 0,01). a, b – Different letters it differs when comparing the results of each
observer in the same variable, rat line, sex and experimental group.
74 ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2013. ¹ 2
D.F. Arencibia-Arrebola, L.A. Rosario-Fernandez, Y.E. Suarez-Fernandez, A.Vidal-Novoa
0.15 ± 0.06 % of polychromatic erythrocytes with
micronucleus. The response of Sprague Daw-
ley line to the ñyclophosphamide was high, but
smaller than the clastogenicity results obtained in
both sexes of Lewis and Wistar lines. The total
MN level in Sprague Dawley rats was lower than
in animals of other lines. On the other hand,
the ñyclophosphamide revealed its biggest clas-
togenic effect in the males of Wistar rats [11,
16]. Significant differences between sexes were
not observed. The different response of rat lines
on ñyclophosphamide is the result of their dif-
ferences in drug metabolism [7, 17, 18]. Íepatic
enzymes are fundamental in the first phase of
xenobiotic metabolism that participates in the
ñyclophosphamide metabolism in the liver [7,
11, 17–21]. The differences in expressions of en-
zymes lead to differences in their metabolisms
[20, 21]; different concentrations of the metabo-
lites, active mutagene levels and so on.
The comparison of data of three independent
observers is shown in the Table 2. In general the
results demonstrated little variation of the vari-
ables and studied experimental groups. These
data are in good correspondence with results of
other investigators [23, 24] where high repeata-
bility and consistency in their micronucleus assay
in bone marrow cells were demostrated. Thus, it
may be recommended to explore genotoxic ef-
fects of drugs at the chromosome level [9].
Being based on the data obtained, we sup-
pose that the Sprague Dawley rat line is more
genetically stable than the other lines. This line
revealed the lower spontaneous indexes and ac-
ceptable cyclophosphamide-induced indexes [25,
26]. These results are important also due to het-
erogeneity of response of the Sprague Dawley
rats, similar to those in human populations [25,
26]. Futher, we suggest to use Wistar rats in ex-
periments with cyclophosphamide intraperitoneal
treatment as positive control. In our experiments
Wistar line was the most sensitive to the muta-
genic action of cyclophosphamide and this line
may be good biomodel to detection of mutagenic
effects of similar substances.
Conclusions. The Sprague Dawley rats of both
sexes differ significantly from the other lines in
lower spontaneous indexes and acceptable in-
duced with cyclophosphamide indexes in the
bone marrow cells micronucleus assay.
The authors thank to the technical personnel of
the animal facilities installation from Research and
Development Vicepresidency of the Finlay Institute,
Cuba.
D.F. Arencibia-Arrebola, L.A. Rosario-Fernandez,
Y.E. Suarez-Fernandez, A. Vidal-Novoa
ÑÐÀÂÍÅÍÈÅ ×ÀÑÒÎÒÛ ÌÈÊÐÎßÄÅÐ
 ÊËÅÒÊÀÕ ÊÎÑÒÍÎÃÎ ÌÎÇÃÀ
ÒÐÅÕ ËÈÍÈÉ ÊÐÛÑ
Ñðàâíèâàëè ñïîíòàííûå è èíäóöèðîâàííûå
öèêëîôîñôàìèäîì ìèêðîÿäåðíûå èíäåêñû â êëåò-
êàõ êîñòíîãî ìîçãà òðåõ ëèíèé êðûñ – Sprague
Dawley, Ëüþèñ è Âèñòàð. Ñôîðìèðîâàëè ïÿòü
ýêñïåðèìåíòàëüíûõ ãðóïï (ïî 10 æèâîòíûõ êàæ-
äîãî ïîëà è ëèíèè â êàæäîé ãðóïïå). Ïåðâóþ ãðóï-
ïó èñïîëüçîâàëè â êà÷åñòâå íåãàòèâíîãî êîíò-
ðîëÿ (èíòàêòíûå æèâîòíûå), âòîðàÿ ñëóæèëà êîíò-
ðîëåì âëèÿíèÿ ïåðîðàëüíîãî ïðèåìà ïðåïàðàòîâ.
Âñå óñëîâèÿ ñîäåðæàíèÿ áûëè îäèíàêîâûìè äëÿ
âñåõ ãðóïï æèâîòíûõ. Êðûñ òðåòüåé ãðóïïû îá-
ðàáàòûâàëè 2 % Tween 65, à ÷åòâåðòîé ãðóïïû –
0,9 % NaCl. Îáà âåùåñòâà ââîäèëè ïåðîðàëüíî â
äîçå 2 ìë/êã â òå÷åíèå 14 äíåé. Æèâîòíûõ ïÿ-
òîé ãðóïïû îáðàáàòûâàëè âíóòðèáðþøèííî öèêëî-
ôîñôàìèäîì â äîçå 50 ìã/êã (10 ìë/êã ðàñòâîðà),
çà 48 è 24 ÷ äî ýâòàíàçèè. Ëèíèÿ Sprague Dawley
(îáà ïîëà) ñóùåñòâåííî îòëè÷àåòñÿ îò äðóãèõ.
Êðûñû ýòîé ëèíèè èìåëè íèçêèå ïîêàçàòåëè
ñïîíòàííîãî îáðàçîâàíèÿ ìèêðîÿäåð è âûñîêèé
èíäåêñ ïðè èíäóêöèè öèêëîôîñôàìèäîì, à òàêæå
ïðîöåíò ýðèòðîöèòîâ ñ ìèêðîÿäðàìè â êîñòíîì
ìîçãå è èíäåêñ öèòîòîêñè÷íîñòè. Ïîëó÷åííûå ðå-
çóëüòàòû ïîçâîëÿþò îïðåäåëèòü íàèáîëåå ïîäõî-
äÿùèå ëèíèè êðûñ â êà÷åñòâå ìîäåëüíûõ æè-
âîòíûõ äëÿ èçó÷åíèÿ ãåíîòîêñè÷íîñòè. Ýòî òàêæå
ïîçâîëèò ïîëó÷àòü áîëåå òî÷íûå îöåíêè ãåíîòîê-
ñè÷íîñòè ðàçëè÷íûõ âåùåñòâ.
D.F. Arencibia-Arrebola, L.A. Rosario-Fernandez,
Y.E. Suarez-Fernandez, A. Vidal-Novoa
ÏÎвÂÍßÍÍß ×ÀÑÒÎÒÈ Ì²ÊÐÎßÄÅÐ
 Ê˲ÒÈÍÀÕ Ê²ÑÒÊÎÂÎÃÎ ÌÎÇÊÓ
ÒÐÜÎÕ Ë²Í²É ÙÓвÂ
Ïîð³âíþâàëè ñïîíòàíí³ òà ³íäóêîâàí³ öèêëî-
ôîñôàì³äîì ì³êðîÿäåðí³ ³íäåêñè â êë³òèíàõ ê³ñò-
êîâîãî ìîçêó òðüîõ ë³í³é ùóð³â – Sprague Daw-
ley, Ëþ³ñ ³ Âèñòàð. Ñôîðìóâàëè ï’ÿòü åêñïåðè-
ìåíòàëüíèõ ãðóï (ïî 10 òâàðèí êîæíî¿ ñòàò³ ³
ë³í³¿ â êîæí³é ãðóï³). Ïåðøó ãðóïó âèêîðèñòàëè
ÿê íåãàòèâíèé êîíòðîëü (³íòàêòí³ òâàðèíè), äðóãà
ñëóãóâàëà êîíòðîëåì âïëèâó ïåðîðàëüíîãî ïðè-
éîìó ïðåïàðàò³â. Âñ³ óìîâè óòðèìàííÿ áóëè îäíà-
êîâèìè äëÿ âñ³õ ãðóï òâàðèí. Ùóð³â òðåòüî¿ ãðóïè
75
Comparison of micronuclei frequency in bone marrow cells of three rat lines
ISSN 0564–3783. Öèòîëîãèÿ è ãåíåòèêà. 2013. ¹ 2
îáðîáëÿëè 2 % Òween 65, à ÷åòâåðòî¿ ãðóïè –
0,9 % NaCl. Îáèäâ³ ðå÷îâèíè ââîäèëè ïåðîðàëüíî
â äîç³ 2 ìë/êã âïðîäîâæ 14 äí³â. Òâàðèí ï’ÿòî¿ ãðóïè
îáðîáëÿëè âíóòð³øíüî÷åðåâíî öèêëîôîñôàì³äîì
â äîç³ 50 ìã/êã (10 ìë/êã ðîç÷èíó), çà 48 ³ 24 ãîä äî
åâòàíà糿. ˳í³ÿ Sprague Dawley (îáèäâ³ ñòàò³) ³ñòîò-
íî â³äð³çíÿëàñü â³ä ³íøèõ. Ùóðè ö³º¿ ë³í³¿ ìàëè
íèçüê³ ïîêàçíèêè ñïîíòàííîãî óòâîðåííÿ ì³êðî-
ÿäåð ³ âèñîêèé ³íäåêñ ïðè ³íäóêö³¿ öèêëîôîñ-
ôàì³äîì, à òàêîæ â³äñîòîê åðèòðîöèò³â ç ì³êðî-
ÿäðàìè â ê³ñòêîâîìó ìîçêó òà ³íäåêñ öèòîòîê-
ñè÷íîñò³. Îòðèìàí³ ðåçóëüòàòè äîçâîëÿþòü âèçíà-
÷èòè íàéá³ëüø â³äïîâ³äí³ ë³í³¿ ùóð³â ÿê ìîäåëüíèõ
òâàðèí äëÿ âèâ÷åííÿ ãåíîòîêñè÷íîñò³. Öå òàêîæ
äîçâîëèòü îòðèìóâàòè òî÷í³ø³ îö³íêè ãåíîòîêñè÷-
íîñò³ ð³çíèõ ðå÷îâèí.
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/HRV (Za stvaranje Adobe PDF dokumenata najpogodnijih za visokokvalitetni ispis prije tiskanja koristite ove postavke. Stvoreni PDF dokumenti mogu se otvoriti Acrobat i Adobe Reader 5.0 i kasnijim verzijama.)
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/NLD (Gebruik deze instellingen om Adobe PDF-documenten te maken die zijn geoptimaliseerd voor prepress-afdrukken van hoge kwaliteit. De gemaakte PDF-documenten kunnen worden geopend met Acrobat en Adobe Reader 5.0 en hoger.)
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| id | nasplib_isofts_kiev_ua-123456789-126533 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 0564-3783 |
| language | Russian |
| last_indexed | 2025-12-07T16:26:18Z |
| publishDate | 2013 |
| publisher | Інститут клітинної біології та генетичної інженерії НАН України |
| record_format | dspace |
| spelling | Arencibia-Arrebola, D.F. Rosario-Fernandez, L.A. Suarez-Fernandez, Y.E. Vidal-Novoa, A. 2017-11-25T19:26:06Z 2017-11-25T19:26:06Z 2013 Comparison of micronuclei frequency in bone marrow cells of three rat lines / D.F. Arencibia-Arrebola, L.A. Rosario-Fernandez, Y.E. Suarez-Fernandez, A. Vidal-Novoa // Цитология и генетика. — 2013. — Т. 47, № 2. — С. 70-76. — Бібліогр.: 26 назв. — англ. 0564-3783 DOI: 10.3103/S0095452713020023 https://nasplib.isofts.kiev.ua/handle/123456789/126533 The aim of this paper is to compare the spontaneous and induced with cyclophosphamide micronucleus indexes in bone marrow cells of the Sprague Dawley, Lewis and Wistar rat lines. Five experimental groups were formed (10 animals of each sex and of each line, in every group). The first group was used as the negative control (intact animals), the second one was exposed to oral administration of drugs; other conditions were the same as for the other groups. The third group was treated with 2 % Tween 65 and the fourth group was treated with 0.9 % NaCl. Both substances were administered by oral way to 2 ml/kg during 14 days. The fifth group was treated intraperitoneally with strong mutagen cyclophosphamide in the dose of 50 mg/kg (10 ml/kg in solution), on 48th and 24th hours before euthanasia. The Sprague Dawley line (both sexes) was significantly different from the other lines. Rats of this line had lower index of spontaneous formation of micronuclei, higher index of cyclophosphamideinduced micronucle formation, percent of micronucleated erythrocytes in bone marrow and the index of cytotoxicity. The results obtained make it possible to identify the most appropriate line of rats as model animals for studies of genotoxicity. It will allow also to obtain more accurate estimates of genotoxicity of various substances. Сравнивали спонтанные и индуцированные циклофосфамидом микроядерные индексы в клетках костного мозга трех линий крыс – Sprague Dawley, Льюис и Вистар. Сформировали пять экспериментальных групп (по 10 животных каждого пола и линии в каждой группе). Первую группу использовали в качестве негативного контроля (интактные животные), вторая служила контролем влияния перорального приема препаратов. Все условия содержания были одинаковыми для всех групп животных. Крыс третьей группы обрабатывали 2 % Tween 65, а четвертой группы – 0,9 % NaCl. Оба вещества вводили перорально в дозе 2 мл/кг в течение 14 дней. Животных пятой группы обрабатывали внутрибрюшинно циклофосфамидом в дозе 50 мг/кг (10 мл/кг раствора), за 48 и 24 ч до эвтаназии. Линия Sprague Dawley (оба пола) существенно отличается от других. Крысы этой линии имели низкие показатели спонтанного образования микроядер и высокий индекс при индукции циклофосфамидом, а также процент эритроцитов с микроядрами в костном мозге и индекс цитотоксичности. Полученные результаты позволяют определить наиболее подходящие линии крыс в качестве модельных животных для изучения генотоксичности. Это также позволит получать более точные оценки генотоксичности различных веществ. Порівнювали спонтанні та індуковані циклофосфамідом мікроядерні індекси в клітинах кісткового мозку трьох ліній щурів – Sprague Dawley, Люіс і Вистар. Сформували п’ять експериментальних груп (по 10 тварин кожної статі і лінії в кожній групі). Першу групу використали як негативний контроль (інтактні тварини), друга слугувала контролем впливу перорального прийому препаратів. Всі умови утримання були однаковими для всіх груп тварин. Щурів третьої групи обробляли 2 % Тween 65, а четвертої групи – 0,9 % NaCl. Обидві речовини вводили перорально в дозі 2 мл/кг впродовж 14 днів. Тварин п’ятої групи обробляли внутрішньочеревно циклофосфамідом в дозі 50 мг/кг (10 мл/кг розчину), за 48 і 24 год до евтаназії. Лінія Sprague Dawley (обидві статі) істотно відрізнялась від інших. Щури цієї лінії мали низькі показники спонтанного утворення мікроядер і високий індекс при індукції циклофосфамідом, а також відсоток еритроцитів з мікроядрами в кістковому мозку та індекс цитотоксичності. Отримані результати дозволяють визначити найбільш відповідні лінії щурів як модельних тварин для вивчення генотоксичності. Це також дозволить отримувати точніші оцінки генотоксичності різних речовин. The authors thank to the technical personnel of the animal facilities installation from Research and Development Vicepresidency of the Finlay Institute, Cuba. ru Інститут клітинної біології та генетичної інженерії НАН України Цитология и генетика Оригинальные работы Comparison of micronuclei frequency in bone marrow cells of three rat lines Сравнение частоты микроядер в клетках костного мозга трех линий крыс Порівняння частоти мікроядер в клітинах кісткового мозку трьох ліній щурів Article published earlier |
| spellingShingle | Comparison of micronuclei frequency in bone marrow cells of three rat lines Arencibia-Arrebola, D.F. Rosario-Fernandez, L.A. Suarez-Fernandez, Y.E. Vidal-Novoa, A. Оригинальные работы |
| title | Comparison of micronuclei frequency in bone marrow cells of three rat lines |
| title_alt | Сравнение частоты микроядер в клетках костного мозга трех линий крыс Порівняння частоти мікроядер в клітинах кісткового мозку трьох ліній щурів |
| title_full | Comparison of micronuclei frequency in bone marrow cells of three rat lines |
| title_fullStr | Comparison of micronuclei frequency in bone marrow cells of three rat lines |
| title_full_unstemmed | Comparison of micronuclei frequency in bone marrow cells of three rat lines |
| title_short | Comparison of micronuclei frequency in bone marrow cells of three rat lines |
| title_sort | comparison of micronuclei frequency in bone marrow cells of three rat lines |
| topic | Оригинальные работы |
| topic_facet | Оригинальные работы |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/126533 |
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