PEER REVIEWED
In light of the various nuclear power plant
accident, military and terrorism scenarios that have entered the world
consciousness over the past two generations, there is tremendous practical
value to understanding how to neutralize or reduce the damaging (and lethal) effects
of radiation on the body. Despite substantial research over the past three
decades, no chemical compounds have been found to be perfectly safe and
effective for this purpose. Research on the use of plant extracts to protect
against radiation exposure is not widely known, however the low toxicity and
minimal side effects of many plant products are well known in both western science
and traditional medicine. Examining the benefits of plant-based therapy with
rigorous quantitative science is beginning to gain a toehold in the field of
radioprotection. There are now a number of mechanism-based reports of
substantial in vitro and in vivo activity from plant
preparations [reviewed in March 2007: Jagetia (2007) Journal of Clinical
Biochemistry and Nutrition. 40: 74-81]. By presenting the following
peer-reviewed study, we hope to add to that body of evidence and to stimulate
further research in this neglected arena.
Abstract
The present investigation reports the radiomodulatory
effect of Rosmarinus officinalis (rosemary) leaf extract against
radiation-induced hematological alterations in Swiss albino mice at various
post-autopsy intervals (i.e., between 24 hours to day 30). Treatment of animals
with rosemary extract (1000 mg/ kg body wt) prior to irradiation was found to
delay the onset of mortality and reduced the symptoms of radiation sickness
such as ruffled hairs, lethargy, anorexia and diarrhea in comparison to
radiation alone treated animals. Rosemary treated experimental groups exhibited
a dose dependent rise (9 < 6 < 3 Gy) in the number of leucocytes (i.e.,
lymphocytes, monocytes, basophils, eosinophils and neutrophils) by the 30th day
post autopsy interval in comparison to the control. Irradiation resulted in a
significant increase in lipid peroxidation levels (p< 0.01, p< 0.001) and
a reduction in glutathione levels (p<0.05, p<0.001) in blood as observed
in radiation alone treated animals. Conversely, treatment of mice with rosemary
extract exhibited a significant decrease (p< 0.01, p< 0.001) in lipid
peroxidation level and an increase (p< 0.05, p<0.001) in glutathione
content.
Introduction
Radiation protection is at a cross-road after
radiation incidents and unacceptable tragedies such as those at Chernobyl and Three
Mile Island. Radiation induced damage to the normal tissues can be partially
reduced by the use of radioprotectors that reduce the damaging effects of
radiation, including radiation-induced lethality (4,22,38). Various workers
have investigated the potential application of radioprotective chemicals in the
event of planned and unplanned exposure (i.e., clinical oncology, radiation
site cleanup, military scenarios, radiological terrorism, radiation accidents,
etc.) (18,25,40).
A substantial amount of research has been carried out
in the field of chemical radioprotection during the last few decades; however,
no safe and ideal synthetic radioprotectors are available to date. Recently,
interest has generated in developing the potential drugs of plant origin for
the amelioration of radiation effects. Plants and their products are well known
to have an advantage over the synthetic compounds in terms of their potential
low/no toxicity at the effective dose with minimum or no side effects (3,9,33,34,41).
However, the use of medicinal plants suffers from lack of robust scientific
evidence to support their use. Therefore, studies supporting or rejecting their
role in the treatment of various health disorders is of great need (3).
Rosemary (Rosmarinus officinalis), belonging to
the family Lamiaceae, is a common medicinal and aromatic plant grown in many
parts of the world. It is indigenous to Southern Europe, particularly on the
dry rocky hills of the Mediterranean region. Rosemary is used as a culinary
herb, a beverage drink, as well as in cosmetics; in folk medicine it is used as
a tonic and stimulant, analgesic, antirheumatic, carminative, diuretic, expectorant,
anti-epileptic, anti-spasmodic in renal colic, dysmenorrhoea, for relief of
respiratory disorders, effects on human fertility, and the stimulation of hair
growth (1). Rosemary has been shown to be safe in toxicity studies in animal
models when added as an antioxidant to food (35).
Since time immemorial, the plant has been used
traditionally by people for curing various health disorders around the world.
Caribs of Guatemala use rosemary to cure various human diseases (12). Rosemary
has been described as a wonder-drug in literature and in various medieval drug
monographs as well (36,44). Thus, wide acceptability and diverse
pharmacological and anti-oxidative properties of the plant stimulated us to
evaluate the radio modulatory effect of Rosmarinus officinalis in Swiss
albino mice exposed to various doses of gamma radiation.
Materials and Methods
Animal care and handling
Male Swiss albino mice (Mus musculus), 6-8
weeks old, weighing 20-24 g., from an inbred colony were used for the present
study. The animals were provided standard mice feed (procured from Hindustan
Lever Ltd., India) and water ad libitum and were maintained under controlled
conditions of temperature and light (Light: dark, 10 hrs: 14 hrs.). Four
animals were housed in a polypropylene cage with locally procured paddy husk (Oryza
sativa) as bedding throughout the experiment. Tetracycline-containing water
(0.13 mg/ml) was provided once a fortnight and was given as a preventive
measure against infections. Animal care and handling were performed according
to the guidelines set by the World Health Organization (WHO), Geneva, Switzerland
and the INSA (Indian National Science Academy), New Delhi, India. The
Departmental Animal Ethical Committee approved the present study.
Irradiation
The cobalt teletherapy unit (Co-60) at the Cancer
Treatment Centre, Radiotherapy Department, SMS Medical College & Hospital, Jaipur,
India, was used for irradiation. Unanaesthetized animals were restrained in
well-ventilated Perspex boxes and exposed to various doses of gamma radiation
(i.e., 3, 6 and 9 Gy) at a distance (SSD) of 80 cm from the source at a dose
rate of 0.85 Gy/min.
Taxonomic description of the plant
Rosemary is an evergreen shrub growing to 1.5 m by 1.5
m at a medium rate. The leaves of rosemary are about 1 inch long, linear,
revolute, dark green above and paler and glandular beneath, with camphoraceous
aromatic odour. The scented hermaphrodite flowers are small and pale blue. Much
of the active volatile principle resides in their calyces. There are various
other varieties of the plant, but the green-leaved variety is the kind used
medicinally (13).
Preparation of plant extract
The identification of the plant Rosmarinus
officinalis (family: Lamiaceae) was done by a botanist, Dr Deepak Acharya,
(Voucher Specimen No: DDC/2001/DEPTBT/ACHARYA2430) of the Department of
Botany, Danielson College, Chhindwara, Madhya Pradesh (India). The non-infected
leaves of the plant were collected, carefully cleaned, shade dried and powdered
in a grinder. The plant material was prepared by extracting 200 gm of leaf
powder with double distilled water by refluxing for 36 hrs (12 hrs. x 3) at 55
± 5°C. Pellets of the extract were obtained by evaporation of its liquid
contents in the incubator. An approximate yield of 22 % extract (w/w) was
obtained.
The required dose for treatment was prepared by
dissolving the drug pellets in double distilled water and administered by oral
gavage with a micropipette (100 µl/ animal) at a dose of 1000 mg/ kg body
wt./animal (1000 mg of 22% of original plant weight). Henceforth, rosemary leaf
extract will be called RE.
Experimental Design
Optimum dose determination
A dose selection of Rosmarinus officinalis (RE)
was done on the basis of a drug tolerance study. For this purpose, various
doses of RE extract (100, 200, 400, 800, 1000, 1500 and 2000 mg/kg body wt.)
were tested for their tolerance (once in a day for 5 consecutive days) in Swiss
albino mice. One hour after the last administration of RE, mice were exposed to
8 Gy gamma irradiation. All these animals were then observed for 30 days for
scoring signs of radiation sickness or mortality. Thus, the optimum tolerated
dose of RE (1000 mg/ kg b. wt.) was determined and used for further detailed
experimentation (Fig 1).
The LD50/30 and Dose Reduction Factor
The efficacy of any protective agent is evaluated by
the determination of its dose reduction factor (DRF). The DRF of R.
officinalis extract (RE) based on LD50/30 survival experiment
was calculated after irradiating a large number of Swiss albino mice to
different doses of gamma rays in the presence (experimental) or absence
(control) of RE. The percentage of mice surviving at each radiation dose till
30 days following such exposures was used to construct survival dose response
curves. Regression analysis was done to obtain LD50/30, and dose
reduction factor was computed as:
DRF =
|
LD50/30 of
Experimental Animals
|
LD50/30 of
Control Animals
|
The LD50/30 values for control and
experimental animals obtained from the survival data are 6.85 and 10.47
respectively. The dose reduction factor of R. officinalis against
radiation treatment was calculated on the basis of the survival experiment and was
measured as 1.53.
Modification of radiation response
A total of 48 animals used for the experiment were assorted into 4 groups. Mice of group 1 (sham irradiated)
were orally administered double distilled water (DDW) at a dose of 1000 mg/ kg
body weight, volume equal to RE. Animals belonging to group 2 (RE alone) were
given daily rosemary extract at a dose of 1000 mg/ kg/ animal for 5 consecutive
days, one hour before irradiation. Animals of group 3 (radiation control) were
exposed to various doses of gamma rays alone (i.e., 3, 6 and 9 Gy) one hour
after DDW treatment on day 5. Group 4 (RE experimental) received RE (1000 mg/
kg body wt./ animal) as in group 2. One hour after last administration of RE,
mice were exposed to various doses of gamma rays, (i.e., 3, 6 and 9 Gy),
respectively. These animals were observed daily for any sign of sickness,
morbidity, behavioral toxicity and mortality. A minimum of 6 animals from each
group were necropsied on days 1, 3, 5, 10, 20 and 30 post-treatment intervals
to study hematological and biochemical parameters.
Hematological study
For the study, blood was collected from the orbital
sinus of animals from each group in a vial containing 0.5 M EDTA. Differential
leucocyte counts (lymphocytes, monocytes, basophils, eosinophils and neutrophils)
were determined by adopting standard procedures. The number of each type of
leucocytes (lymphocytes, monocytes, basophils, eosinophils and neutrophils) and
the total number of leucocytes counted were recorded. The percentage of each
type of leucocyte was calculated by the formula:
Number of
type of leucocyte
|
× 100
|
Total number
of leucocytes counted
|
Biochemical determinants
Biochemical alterations were studied in animals of all
the groups at one hour post- exposure to gamma radiation. The level of
glutathione (GSH) was determined in blood by the method of Beutler et al.
(5). The lipid peroxidation (LPx) level in the serum was measured by the assay
of thiobarbituric acid reactive substances (TBARS) according to the method of
Okhawa et al. (30).
Statistical
analysis
The result for all the groups at various necropsy
intervals were expressed as mean ± standard error of the mean (S. E. M.) to
evaluate whether the mean of the sample drawn from experimental (RE
experimental) deviated significantly from respective control (Irradiation
control). Student’s ‘t’ test was used by the method of Bourke et al. (7).
The significance level was set at different levels as p<0.05, p<0.01 and
p<0.001.
Results
Data are presented in Tables 1-3 and Figures 1-4. The
radioprotective effect of rosemary leaf extract (RE) was studied in mice
treated with 1000 mg/ kg body wt. RE before exposure to 3, 6 and 9 Gy of gamma
radiation. No noticeable signs of behavioral change, sickness or mortality were
observed in Sham irradiated/ RE-treated group. Animals exposed to 3 and 6 Gy
gamma rays alone did not show mortality throughout the experimental period, but
slight laziness was observed in some animals. Animals exposed to 9 Gy gamma
rays exhibited epilation, ruffled hair, watering of eyes, weight loss and
became lethargic. No animal could survive in the 9 Gy irradiated alone group
beyond day 10. Animals pretreated with RE did not exhibit mortality or any
symptoms of radiation sickness. General health, activeness, food and water
intake were found to be normal in RE pretreated irradiated animals.
After whole body exposure to different doses of gamma
radiation (i.e., 3, 6 and 9 Gy), lymphocyte percentages remained significantly
lower than normal, and could not regain a normal value even by the last day of
autopsy interval (day 30). No significant changes in monocytes, eosinophils and
basophil counts were registered in any of the groups. However, monocytes
followed a pattern similar to lymphocytes. Following irradiation, a significant
increase above normal was observed in neutrophil counts. A normal value could
not be restored in any of the irradiated groups till day 30 post-exposure.
Daily administration of 1000 mg/ kg of RE for 5
consecutive days rendered recovery in the different types of leucocytes (i.e.,
lymphocytes, monocytes, eosinophils, basophils and neutrophils) in comparison
to irradiated alone groups, and values close to normal were registered in a
dose dependent manner (3> 6> 9 Gy) by post-treatment day 30.
Biochemical determinants
There was no significant difference observed in the
levels of glutathione (GSH) and lipid peroxidation (LPx) in the blood content
of sham irradiated (group 1) or RE alone treated animals (group 2). In
concomitant treatment of RE and radiation (group 4), GSH was found
to be further lowered than the radiation treated group. A significant elevation
in the values of blood GSH as compared to group 3 was estimated in RE
experimental animals. An increase in LPx levels above normal was evident in
serum of irradiated mice, while a significant decrease in such values was
evident in the RE pretreated irradiated group.
Discussion
Radiation injuries are manifested as a result of
enhanced production of free radicals due to oxidative stress. Exposure to
radiation causes ionization of molecules in the cells, which sets off
potentially damaging reactions via free radical production (23). Free
radical mediated processes and oxidative stress have been implicated in the
pathogenesis of the aging process and various diseases such as atherosclerosis,
liver damage, arthritis, cancer, and neurodegenerative disorders (2). The
prevailing view is that intake of antioxidant nutrients can reduce the risk of
free radical-related health problems and may prove to be protective against
ionizing irradiation.
The present study revealed that the number of
lymphocytes declined in a dose dependent manner after exposure to 3, 6 and 9 Gy
gamma irradiation. A rapid depression was observed at early intervals which may
be attributed to direct destruction of such cells in peripheral blood of mice (31).
No significant changes in monocytes, eosinophils and basophils were observed
after whole body exposure to different doses of gamma radiation. It may be
attributed to the fact that mature granulocytes are radioresistant whereas
lymphocytes are extremely sensitive to radiation (8). Furthermore, neutrophil
granules altered inversely as compared to lymphocytes. These cells exhibited an
early rise while lymphocytes and monocytes declined soon after exposure thus
showing an opposite behavior. This can be explained by an abortive rise
phenomenon as described earlier by workers (26,39).
According to Hall (14), by the time the number of
circulating cells in the blood reaches minimum value as the mature circulating
cells begin to die off, the supply of new cells from the depleted precursor
population becomes inadequate to replace these, thereby making radiation
effects become apparent. Also, this abrupt increase may have appeared due to an
abortive rise phenomenon (6,26) or can be interpreted as stimulation effect (11).
Hastening the maturation of granulocyte precursors in bone marrow and their
release into general circulation can be attributed to a rise in neutrophil
counts (28,42).
It is evident from the present study that administration
of RE reduced radiation sickness and mortality, and provided protection to
differential leucocytes counts (i.e., lymphocytes, monocytes, basophils,
eosinophils and neutrophils) in the peripheral blood of mice from the damaging
gamma radiation. It has been observed that rosmarinic acid (found in rosemary)
is effective in relation to blood circulation and to improve hemodynamics in
occlusive arterial diseases (1). Rosemary has been found to contain certain
antioxidative (32) and free radical scavenging activity (45) in its active
compounds like caffeic acid, carnosolic acid, chlorogenic acid, rosmanol,
rosmarinic acid, carnosol, different diterpenes (16,43), rosmari- diphenol,
rosmariquinone (17) and other natural antioxidants such as ursolic acid,
alkaloid rosmaricine and glucocolic acid (21). In a recent study, carnosic acid
was found to render protection to UVA irradiated human skin fibroblasts (29).
The basic effect of radiation on cellular membranes is
believed to induce lipid peroxidation (LPx) by the production of free radicals
that have the potential to damage DNA and cause cell death (24,27). The level
of radiation-induced LPx increased considerably in a dose dependent manner in
the entire group 3 irradiated animals, whereas a decrease in the values was
observed in RE-treated group 4. The inhibition observed in the LPx level in
blood of RE administered animals may have been responsible for the observed
radioprotection by plant extract. This view is supported by the investigation of
an anti-lipoperoxidant activity of young sprouts of Rosmarinus officinalis
that significantly reduced the formation of malondialdehyde in rat hepatocytes1
(9). Sotelo-Felix et al. proposed that carnosol could scavenge free
radicals induced by carbon tetrachloride, consequently avoiding the propagation
of lipid peroxides in the liver of mice (37).
Studies conducted by Haraguchi et al. report an
inhibition of superoxide and lipid peroxidation by 4 diterpenoids from
rosemary, i.e. carnosic acid, carnosol, rosmanol and epirosmanol (15). Del Bano
et al. investigated the efficacy of carnosic acid, carnosol and
rosmarinic acid (active constituents of rosemary) and found these to be
radioprotective against chromosomal damage induced by γ-rays (10).
The exact mechanism of action of rosemary is yet to be elucidated; however, it
may act as a protective by scavenging free radicals triggered by radiation.
Glutathione (GSH) is one of the antioxidant enzymes
that act as the first line of defense against pro-oxidant stress, thus
performing as a free radical scavenger. Oral administration of DDW or RE did
not significantly influence the endogenous GSH level in blood. In the present
study, GSH levels were found to be lower in the blood of irradiated alone
animals than that observed in the RE pre-treated mice. The levels of GSH were
found to be elevated in the blood of mice after RE administration.
One of the mechanisms of RE protection against
radiation can be an elevation in the glutathione level that is mediated through
the modulation of cellular antioxidant level. Rosmarinic acid has been
experimentally found to have a significant antioxidant role through free
radical scavenging activity (1). Kilic et al. observed that lipid
peroxidation starts as soon as the endogenous GSH gets exhausted, and the
addition of GSH stops further peroxidation promptly (20). Increase in the GSH
concentration, towards normal, could have resulted in reduced levels of LPx,
thereby protecting against damage caused by radiation in the RE pre-treated
irradiated group.
The mechanism of the radioprotective action of Rosmarinus
officinalis leaf extract in this animal model may thus be its free radical
scavenging activity and its ability to thus protect cellular molecules from
oxidative damage. Furthermore, it inhibited lipid peroxidation and modulated
GSH levels in blood of these Swiss albino mice. The activity of rosemary may
also be attributed to stimulating or protecting hematopoiesis in bone marrow
and a subsequent increase of hematological constituents in the peripheral
blood. Since significant protection was obtained at a non-toxic low dose, RE
may have an advantage over the known radioprotectors. Further investigations
are in progress to study the exact mechanism of action and clinical
applicability of R. officinalis in radioprotection.
Table 1: Variation in differential leucocyte
counts (DLC) of 3 Gy irradiated Swiss albino mice at various autopsy
intervals
|
Post-Irradiation Interval
|
Treatment Group
|
Lymphocytes
(%)
|
Monocytes
(%)
|
Eosinophils
(%)
|
Basophils
(%)
|
Neutrophils
(%)
|
Day 1
|
Irradiation Control
|
48.4±0.10c
|
2.2±0.50b
|
1.4±0.66
|
0.8±0.33
|
47.2±0.79c
|
RE Experimental
|
54.8±3.16c
|
2.6±0.82
|
2.0±0.21
|
0.6±0.35
|
40.0±1.50c
|
Day 3
|
Irradiation Control
|
52.0±1.56c
|
2.0±0.44c
|
1.8±0.52
|
0.6±0.21
|
43.6±0.91c
|
RE Experimental
|
57.8±1.28b
|
2.4±0.35
|
1.6±0.59
|
0.4±0.17
|
37.8±1.88c
|
Day 5
|
Irradiation Control
|
58.2±1.38b
|
1.6±0.92b
|
2.4±0.21
|
0.4±0.21
|
37.4±1.65c
|
RE Experimental
|
59.6±2.16
|
1.8±1.01
|
2.6±0.40
|
0.8±0.17
|
35.2±1.80
|
Day 10
|
Irradiation Control
|
58.6±1.50b
|
1.8±0.40b
|
1.8±0.53
|
0.4±0.33
|
36.4±1.30c
|
RE Experimental
|
61.6±1077
|
1.4±0.21
|
2.2±0.48
|
0.6±0.35
|
34.2±1.86c
|
Day 20
|
Irradiation Control
|
56.8±3.28c
|
1.6±0.45c
|
2.0±0.33
|
0.8±0.28
|
38.8±1.95
|
RE Experimental
|
59.2±1.92
|
2.4±0.77
|
1.6±0.17
|
0.6±0.33
|
36.2±1.62
|
Day 30
|
Irradiation Control
|
58.8±2.42a
|
1.8±0.87c
|
2.0±0.48
|
0.6±0.21
|
36.0±1.08c
|
RE Experimental
|
64.8±1.34
|
2.8±0.63
|
2.2±0.77
|
0.4±0.17
|
30.2±0.60
|
Sham-irradiated
|
67.2±2.16
|
3.2±0.40
|
2.8±0.17
|
0.8±0.21
|
25.4±0.52
|
RE alone
|
67.4±1.34
|
3.4±0.22
|
3.2±0.54
|
0.8±0.12
|
25.2±0.45
|
|
Table 2: Variation in differential leucocyte
counts (DLC) of 6 Gy irradiated Swiss albino mice at various autopsy
intervals
|
Post-Irradiation Interval
|
Treatment Group
|
Lymphocytes
(%)
|
Monocytes
(%)
|
Eosinophils
(%)
|
Basophils
(%)
|
Neutrophils
(%)
|
Day 1
|
Irradiation Control
|
51.8±1.68c
|
2.4±0.66
|
2.2±0.35a
|
0.2±0.33
|
43.4±1.03c
|
RE Experimental
|
54.4±2.18
|
2.2±0.35
|
2.6±0.66
|
0.6±0.17
|
40.2±1.80b
|
Day 3
|
Irradiation Control
|
53.8±2.77b
|
2.4±0.17a
|
1.8±0.63
|
0.4±0.21a
|
41.6±1.63c
|
RE Experimental
|
51.6±1.46
|
2.2±0.44
|
1.4±0.95
|
0.2±0.35
|
44.6±1.85
|
Day 5
|
Irradiation Control
|
58.2±0.92
|
1.2±0.21c
|
2.6±0.96
|
0.4±0.33
|
39.6±1.10c
|
RE Experimental
|
56.4±1.94
|
1.6±0.33
|
2.2±1.06
|
0.4±0.28
|
39.4±1.79
|
Day 10
|
Irradiation Control
|
50.0±3.35c
|
1.4±0.78a
|
2.2±0.33a
|
0.6±0.21
|
45.8±1.51c
|
RE Experimental
|
57.2±2.60
|
1.0±0.25
|
1.2±0.56a
|
0.4±0.17
|
40.2±0.93b
|
Day 20
|
Irradiation Control
|
54.6±2.14c
|
1.2±0.48b
|
1.8±0.45a
|
0.2±0.35
|
42.2±0.40c
|
RE Experimental
|
57.8±2.60
|
2.2±0.46a
|
2.0±0.48
|
0.8±0.28
|
37.2±0.79c
|
Day 30
|
Irradiation Control
|
52.2±1.66c
|
1.6±0.60a
|
2.0±0.53
|
0.6±0.10
|
43.6±0.80c
|
RE Experimental
|
63.8±1.37
|
1.8±0.43
|
2.0±1.16
|
0.4±0.28
|
31.8±1.03
|
Sham-irradiated
|
67.2±2.16
|
3.2±0.40
|
2.8±0.17
|
0.8±0.21
|
25.4±0.52
|
RE alone
|
67.4±1.34
|
3.4±0.22
|
3.2±0.54
|
0.8±0.12
|
25.2±0.45
|
|
Table 3: Variation in differential leucocyte
counts (DLC) of 9 Gy irradiated Swiss albino mice at various autopsy
intervals
|
Post-Irradiation Interval
|
Treatment Group
|
Lymphocytes
(%)
|
Monocytes
(%)
|
Eosinophils
(%)
|
Basophils
(%)
|
Neutrophils
(%)
|
Day 1
|
Irradiation Control
|
52.0±2.38
|
3.0±0.44
|
2.8±0.21
|
0.2±0.21
|
42.0±0.92
|
RE Experimental
|
54.0±1.32
|
2.0±0.52
|
2.6±0.40
|
0.2±0.17
|
41.2±1.61
|
Day 3
|
Irradiation Control
|
51.8±1.44
|
2.8±0.21
|
2.6±0.17
|
0.2±0.21
|
42.6±2.95
|
RE Experimental
|
53.8±0.96
|
2.0±0.63
|
2.0±0.25
|
0.4±0.43
|
41.8±1.13
|
Day 5
|
Irradiation Control
|
56.0±2.24
|
2.4±0.48
|
3.0±0.35
|
0.4±0.17
|
38.2±2.51
|
RE Experimental
|
58.2±1.86
|
2.6±0.95
|
1.8±0.84
|
0.2±0.35
|
37.2±1.08
|
Day 10
|
Irradiation Control
|
55.8±1.45
|
2.4±0.71
|
2.8±0.65
|
0.8±0.28
|
38.2±2.57
|
RE Experimental
|
66.4±2.56
|
2.2±0.45
|
1.4±1.12
|
0.6±0.33
|
29.4±1.68
|
Day 20
|
Irradiation Control
|
N.S.
|
N.S.
|
N.S.
|
N.S.
|
N.S.
|
RE Experimental
|
65.6±1.73
|
2.2±0.77
|
1.4±1.08
|
0.2±0.21
|
30.4±3.68
|
Day 30
|
Irradiation Control
|
N.S.
|
N.S.
|
N.S.
|
N.S.
|
N.S.
|
RE Experimental
|
64.2±1.16
|
2.0±0.71
|
1.2±0.82
|
0.4±0.17
|
31.4±3.83
|
Sham-irradiated
|
67.2±2.16
|
3.2±0.40
|
2.8±0.17
|
0.8±0.21
|
25.4±0.52
|
RE alone
|
67.4±1.34
|
3.4±0.22
|
3.2±0.54
|
0.8±0.12
|
25.2±0.45
|
|
(Click to enlarge)
Fig 1: Days of survival of mice pretreated with different
doses of RE and exposed to 8 Gy gamma radiation
(Click to enlarge)
Fig 2: Variations in the body weight of mice at
post-irradiation intervals treated with (experimental) or without Rosemary
extract (RE) treatment
(Click to enlarge)
Fig 3: Variations in the glutathione (GSH) and lipid
peroxidation (LPx) level in peripheral blood of mice with / without Rosemary
extract (RE) treatment
(Click to enlarge)
Fig 4: Survival dose-response curve for determination of
LD50/30 (Survival data collected for treatment of radiation doses
and calculated by regression analysis)
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