Radio-protective Potential of Rosemary (Rosmarinus Officinalis) against Effects of Ionising Radiation

Abstract

Several herbs have been screened for their antioxidative activity using chemiluminescence method. Rosemary
contains phenolic diterpenes, phenolic acids and flavonoids which protect cells and their organic constituent
molecules from free radical radiation- induced oxidative damage. Effects of 0.1 and 0.2 g/Kg body wt. of
Rosemary extract on radiation – induced morbidity and mortality in mice exposed to 10Gy of gamma radiation
were studied for the characterization of high efficiency protection exhibited by Rosemary extract.

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Radio-protective Potential of Rosemary (Rosmarinus Officinalis)
against Effects of Ionising Radiation
MARIA MARIS1, DAN A. MARIS1, SILVIU JIPA2,3*, TRAIAN ZAHARESCU3, LAURA MONICA GORGHIU2
1„Ovidius” University of Constanþa, Faculty of Dental Medicine, 7 Ilarie Horonca Str., 900684, Constanta, Romania
2 Valahia University of Targoviste, Faculty of Sciences and Arts, 18-24 Unirii Av., 130082, Targoviste, România
3 INCDIE ICPE CA, Department of Radiation Processing, 313 Splaiul Unirii, 030138, Bucharest, România
Several herbs have been screened for their antioxidative activity using chemiluminescence method. Rosemary
contains phenolic diterpenes, phenolic acids and flavonoids which protect cells and their organic constituent
molecules from free radical radiation- induced oxidative damage. Effects of 0.1 and 0.2 g/Kg body wt. of
Rosemary extract on radiation – induced morbidity and mortality in mice exposed to 10Gy of gamma radiation
were studied for the characterization of high efficiency protection exhibited by Rosemary extract.
Keywords: rosemary, chemoluminescence, antioxidative activity, ionizing radiation, morbidity, mortality
The human organisms could be exposed to ionizing
radiation in clinical, military or industrial applications.
Gamma irradiation causes oxidative stress generating
reactive oxygen species such as hydroxyl (HO·) radicals,
superoxide radicals (O2 ) and hydrogen peroxide (H2O2).
These free radicals can easily damage the structural and
functional components of cells such as lipids, proteins and
nucleic acids, causing oxidative injury and disturbance in
cellular metabolism. By definition, radioprotectors are
chemical compounds that have the ability to reduce the
oxidative effects of ionizing radiation on normal tissues.
Numerous studies have examined the radio-protective
effects of antioxidant compounds, generally known as free
radical scavengers, which protect cells from free radical
damage. Various plant extracts such as Ginkgo biloba [1],
Panax ginseng [2], Mentha piperita [3], Zingiber officinale
[4], Camellia sinensis [5], Hippophae rhammoides [6],
Allium sativum [7], Agaricus blazei a.s.o., have been
investigated to evaluate their radio-protective effects.
The present study deals with the radio-protective activity
of ethanolic extract prepared from Rosmarinus officinalis
in terms of antioxidative activity and on Swiss albino mice
post irradiation morbidness and death rate, respectively.
Experimental part
The dried Rosemary plant (10 g) and the extracting
solvent (ethanol) were placed in an Erlenmeyer flask
(250mL); the ratio of plant material and extracting solvent
was 1:10 w/v.
Maceration was performed for 120 h at room
temperature, by permanent shaking.
* email: jipasilviu@yahoo.comTel.: (+40) 01 / 3143508 int. 2153
The liquid extract was separated from the plant material
by filtration, and the solvent was evaporated under vacuum.
The solid extract has been used for paraffin modification
(0.25 wt %). Similarly, paraffin samples of extracts from
other radioprotective extracts from plants have been
obtained (fig. 1).
Isothermal chemiluminescence determinations were
performed in air at 1600C by a Lumipol-3 instrument. The
meanings of kinetic that are evaluated in this paper are
presented in figure 1 and table 1.
The radio-protective capability of the Rosemary extract
was analysed on a batch of 60 white mice. All mice were
adults, ages over 15 weeks, both genders, weight ranging
between 20 and 30 g each. The location chosen for the
experiment provided a constant environmental temperature
of 21±2oC, an atmospheric pressure of 759±4 mm Hg
and natural lighting. For a period of four days the mice were
given tetracycline to prevent infection.
Fig. 1. Kinetic parameters from typical chemiluminogram
Table 1
CL PARAMETERS AND THEIR MEANING
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The tested animals were divided into three experimental
batches, each equal in number. The mice batches were
kept in distinct conteiners, marked for identification.
For seven days the experimental batches were subject
to a food diet in the following way:
- batch 1 – was given a standard diet for 7 days;
- batch 2 – the daily diet included and average Rosemary
extract dose of 0.1 g / Kg body wt. for 7 days;
- batch 3 – the mice were given and average Rosemary
extract dose of 0.2 g / Kg body wt. for 7 days.
After seven days, the mice batches were irradiated with
137Cs γ radiation at dose of 10 Gy using a GAMATOR M-38-
2 source. Then, the batches were monitored to observe
the occurrence of morbidity and death rate.
Results and discussions
Figure 2 illustrates the isothermal chemiluminescence
curves (1600C, air) of the paraffin stabilized with various
plant extracts which prove radio-protection effect. The
antioxidant efficiency in stabilizing an organic substrate is
given by the increased value of the induction time as well
as by the lower value of the oxidation rate in comparison
with the unprotected sample. The curve describing the
contribution of Rosemary to the inhibition of substrate
oxidation presents the biggest induction time of the
samples. This feature denotes the exceptional efficiency
of Rosemary extract in the prevention of oxidation.
Table 2 shows the oxidation kinetic parameters of the
samples presented in figure 2.
The longer is the induction time, the stronger is the
antioxidant activity. The analysis of the data in table 2 clearly
shows that the Rosemary extract has the highest
antioxidant activity. The researchers [9] showed that the
antioxidant activity of the Rosemary extract is due, mainly
(over 90%), to some phenolic diterpenes such as carnosic
acid, carnosol, rosmanol, isorosmanol, rosmadial,
epirrosmanol, rosmaridiphenol, rosmariquinone etc (fig. 3)
[9]. Most of these diterpenes act as oxidative chain
breakers.
Carnosic acid has its potent antioxidant activity based
on oxidation cascade (fig. 4): after its molecules interacted
with oxidizing material, it is turned to carnosol. Carnosol
also extracts a free radical becoming rosmanol. Rosmanol
continues the free radical scavenging until galdosol is
formed, and further continues the scavenging process [10,
11].
Fig. 2. Isothermal CL curves (1600C, air) for paraffin stabilized (0.25%
wt%) with various plant extracts which prove a radioprotection effect:
(1) control; (2) Garlic; (3) Ginseng; (4) Agaricus; (5) Mint;
(6) Sea-buckthorn; (7) Ginkgo; (8) Ginger; (9) Green Tea; (10)
Rosemary
Table 2
KINETIC OXIDATION PARAMETERS (CL) FOR PARAFFIN
DOPED WITH VARIOUS RADIOPROTECTIVE PLANT EXTRACTS
Fig. 3. Phenolic diterpenes in Rosemary composition
The caffeic and rosmarinic acids associated with
flavonoids represent other important source of Rosemary
antioxidants.
Ginkgo (Ginkgo biloba)
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The ionizing radiation occurred during exposure induce
the lipid peroxidation chain reactions, which damage DNA
and cell. The phenolic antioxidant contents in the
composition of Rosemary extract, taken before irradiation,
significantly reduce lipid peroxidation [12] (scheme 1).
Another hypothesis considers the increased level of
glutathione in the blood for Rosemary pre-treated animals;
this prevents lipid peroxidation [12-15]. Rosemary extract
proved to be an effective radio-protector in animals
exposed to gamma irradiation dose of 10 Gy.
Figure 4. Oxidation cascade reactions of carnosic acid
Scheme 1 Lipid oxidation
The reactions of these compounds with peroxy radicals
follow the scheme:
The outstanding antioxidant activity of Rosemary
extract is based on its radio-protective capability which
results from the morbidity and mortality analysis of the
mice fed with Rosemary extract and further irradiated with
γ radiation of 137Cs at the dose of 10 Gy. Morbidity watched
out for the pathological skin modifications (hair loss) and
the appearance of toxic signs (diarrhoea) with the mice
batches under study. Due to the increase in the Rosemary
extract amount given to the mice batches, a delay in the
occurrence time of mice morbidity and mortality was
recorded as shown in figure 5.
Fig. 5. The delay in the mice morbidity and mortality after γ 137Cs
(10Gy) irradiation due to the increase of the administrated
Rosemary dose
Conclusions
Rosemary extract shows a remarkable activity in
stabilization of organic substrate.
Rosemary extract could offer protection against the
effects of ionizing radiation because of their ability to
scavenge free radicals.
Phenolic diterpenes, caffeic and rosmarinic acids
associated with flavonoids in Rosemary extract suppress
lipid peroxidation and stop oxidative DNA damage and so
it may be useful as radio-protective agent.
Chemiluminescence has proved its versatility in fast and
accurate assessment degradation study.
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Manuscript received: 22.10.2009