Phytomedicine, Vol.

9: 99108, 2002
Urban & Fischer Verlag
http://www.urbanfischer.de/journals/phytomed

Phytomedicine

The evaluation of the radioprotective effect of Triphala

(an ayurvedic rejuvenating drug) in the mice exposed
to -radiation
G. C. Jagetia1, M. S. Baliga1, K. J. Malagi2 and M. Sethukumar Kamath2
1
2

Department of Radiobiology, Kasturba Medical College, Manipal, India

Department of Ayurveda, Kasturba Hospital, Manipal, India

Summary
The effect of 0, 5, 6.25, 10, 12.5, 20, 25, 40, 50 and 80 mg/kg b. wt. of aqueous extract of triphala (an
Ayurvedic herbal medicine) administrered intraperitoneally was studied on the radiation-induced
mortality in mice exposed to 10 Gy of -radiation. Treatment of mice with different doses of triphala consecutively for five days before irradiation delayed the onset of mortality and reduced the
symptoms of radiation sickness when compared with the non-drug treated irradiated controls.
The highest protection against GI (gastrointestinal) death was observed for 12.5 mg/kg triphala,
where a highest number of survivors were reported up to 10 days post-irradiation. While 10 mg/kg
triphala i.p. provided the best protection as evidenced by the highest number of survivors after 30
days post-irradiation in this group when compared with the other doses of triphala. Toxicity study
showed that triphala was non-toxic up to a dose of 240 mg/kg, where no drug-induced mortality
was observed. The LD50 dose i.p. of triphala was found to be 280 mg/kg b. wt. Our study demonstrates the ability of triphala as a good radioprotective agent and the optimum protective dose of
triphala was 1/28 of its LD50 dose.
Key words: Triphala, radiation, mice, survival, acute toxicity, radioprotection, Terminalia chebula Retz.,
Phyllanthus emblica Linn. or Emblica officinalis Gaertn.and Terminalia bellerica (Gaertn.) Roxb.

 Introduction
The search for radioprotectors started with the realization of the need for a safeguard against the military use
of atomic weapons. With the recognition that normal
tissue protection in radiotherapy is as important as the
destruction of the cancer cells, the focus of protection
research became more therapy oriented. The use of certain chemical agents may reduce the ill effects of radiation in such conditions. Patt et al. (1949) for the first
time observed that the pretreatment of rats and mice
with cysteine before exposure to radiation protected
them against the radiation-induced sickness and mortality. Subsequently, several chemical compounds were
synthesized and tested for their radioprotective ability
(Sweeny, 1979). Only sulphydryl compounds have

been found to be superior radioprotectors than the other

non-sulphydryl compounds but the major drawback of
these compounds has been their high toxicity at the optimum protective dose (Sweeny, 1979), which precluded their effective use in men. A turning point came with
the observation that s-2-(aminopropylamino) ethylphosphorothioic acid (WR-2721) showed substantial
and selective protection of normal tissues with little or
no protection to the solid tumors (Yuhas, 1980). However, it has also been found to be highly toxic at the optimum protective dose and the possibility of using it on
a daily basis was not feasible (Cairnie, 1983). Therefore, it is desirable to search other materials that are
less toxic and can offer high protection.
0944-7113/02/09/02-099 $ 15.00/0

100

G. C. Jagetia et al.

The herbal drugs offer an alternative to the synthetic

compounds and have been considered either non-toxic or
less toxic and this has given impetus to screen for their
radioprotective ability. The mechanism of action of
herbal drugs and their extract preparations differ in many
respects from that of the synthetic drugs or single substances (Wagner, 1999). It can be characterized as a polyvalent action and interpreted as additive or, in some
cases, potentiating. Studies carried out in the past decade
and a half have shown that the herbal preparations like
Liv. 52, protected mice against the radiation-induced
sickness, mortality, dermatitis, spleen injury, liver damage, decrease in the peripheral blood cell counts, prenatal
development, lipid peroxidation and radiation-induced
chromosome damage (Saini et al., 1984a, b; Saini and
Saini, 1985; Saini et al., 1985; Ganapathi and Jagetia,
1995; Jagetia and Ganapathi, 1989, 1991). The brahmarasayana, narasimharasayana, ashwagandharasayana,
and amrithaprasham, a group of herbal preparations used
to improve the general health, have also been reported to
reduce the radiation-induced lipid peroxidation in the
liver, and leucopenia in mice (Kumar et al., 1996).
Abana, an another herbal preparation, clinically used in
India as a cardioprotective agent has also been reported
to protect the mice bone marrow against the radiation-induced micronuclei formation (Jagetia and Aruna, 1997).
According to the Ayurvedic system of medicine, the
body is composed of tridosha or three humours, vata,
translated into wind, corresponds to mind and nervous
system, the pitta translated into fire or bile and is responsible for all metabolic transformations including
digestion and assimilation of the food, while kapha is
translated as water or mucus and it is responsible for
the anabolic functions such as development of muscle
and bone tissues. Triphala, a compound formulation of
the herbs, Terminalia chebula, Phyllanthus emblica or
Emblica officinalis and Terminalia bellerica has been
described in the Ayurveda as a tridoshic rasayan,
having balancing and rejuvenating effects on the three
constitutional elements that govern human life i.e.
vata, pitta and kapha by Charka (1,500B.C.) in the
Charaka Samihita (Sharma and Dash, 1998). In the
Ayurveda, the word rasayana, is a term used for a therapy that produces sturdiness of the body, the sense organs and the teeth, prevent wrinkles in skin, graying of
hair and promote the immune functions and intellect
and render longevity to life (Sharma and Dash, 1998).
Triphala, is one of the important rasayana drugs commonly used in the Ayurvedic system of medicine. This is
an antioxidant rich herbal formulation (Vaidya et al.,
1998; Jose and Kuttan, 1995; Naiwu et al., 1992, Takagi
and Sanashiro, 1996) that has been reported to treat anemia, jaundice, constipation, cough, asthma, fever, eye
diseases, chronic ulcers, leucorrhoea, pyorrhea (Nadkarni, 1976) and also assists in the weight loss

(Hashimoto and Nakajima, 1997). Triphala and/or its individual plant constituents have been reported to possess anti-bacterial (Nadkarni, 1976; Mehta et al., 1993;
Ahmad et al., 1998; Phadke and Kulkarni, 1989; Mehta
et al., 1993), antimalarial (Valsaraj et al., 1997), antifungal activity (Dutta et al., 1998; Valsaraj et al., 1997),
anti-cancer (Tokura and Kagawa, 1995), anti-mutagenic
(Rani et al., 1994; Niwa et al., 1995) anti-allergic (Takagi and Sanashiro, 1996) and anti-viral activities (Valsaraj et al., 1997; Badmaev and Nowakowski, 2000;
Yukaka et al., 1996; Kurokawa and Sato, 1995; Hozumi
and Oyama, 1997; El-Mekkawey and Meselhy, 1995) in
different study systems. Triphala is a cardio-tonic and
exerts its protective effect by improving the blood circulation, reducing the myocardial necrosis (Tariq et al.,
1977), serum cholesterol levels and strengthens the capillaries (Tariq et al., 1977; Hussain, 1975; Thakur, 1984;
Thakur et al., 1988). It is also hepatoprotective and improves the liver function (Gulati et al., 1995; Anand and
Singh, 1997). The decoction of triphala has been found
to treat leucorrhea in women (Singh and Londhe 1993).
It is an effective laxative and improves the gastrointestinal motility (Tamhane et al., 1997) thereby curing the
diseases of gastrointestinal tract (Nadkarni, 1976; Antarkar et al., 1980). Triphala has been reported to possess
anti-aging properties and improves the mental faculties
(Nadkarni, 1976; Antarkar et al., 1980). Triphala has
been found to potentiate the adrenergic function thereby
enabling the body to recover from stress. In addition, the
immunomodulatory property (Suresh and Vasudevan,
1994; Rege et al., 1999) may help in increasing the
bodys defence system resulting in the enhancement of
the body resistance against the diseases (Nadkarni,
1976). The diverse medicinal properties attributed to
triphala and its antioxidant properties stimulated us to
investigate the radioprotective activity of triphala.
The lesson from the experience with radioprotectors
world wide is that the animal studies with death as the
end point is the most confirmatory, because the 30 days
time period after lethal whole body irradiation clearly indicates the capacity of the drug, in test to modulate the recovery and regeneration of the gastrointestinal epithelium and the hemopoietic progenitor cells in the bone marrow, the two most radiosensitive organs that are essential
for sustaining of the life. The aim of the present study was
to evaluate the radioprotective effect of various doses of
triphala in the mice exposed to 10 Gy of whole-body
gamma radiation taking survival as the end point.

 Materials and methods

The animal care and handling was done according to the
guidelines set by the World Health Organization, Geneva, Switzerland and the INSA (Indian National Science

The evaluation of the radioprotective effect of Triphala

Academy, New Delhi, India). Eight to ten week old male
Swiss albino mice weighing 30 to 36 g were selected
from an inbred colony maintained under the controlled
conditions of temperature (23 2 C), humidity (50
5%) and light (10 and 14 h of light and dark, respectively). The animals were provided with the sterile food
and water ad libitum. Four animals were housed in a
polypropylene cage containing sterile paddy husk (procured locally) as bedding throughout the experiment.
Composition of the drug

As the name indicates, triphala (tri = three, phala =

fruits) is a mixture of fruits of three plants namely Terminalia chebula Retz. (Family Combretaceae), Terminalia bellerica (Gaertn.) Roxb. (Family Combretaceae) and Phyllanthus emblica Linn or Emblica officinalis Gaertn. (Euphorbiaceae) in powdered form in
equal proportions (1:1:1).
Preparation of the extract

The aqueous extract of different batches of triphala powder was prepared as described in the Ayurvedic text.
Briefly, 100 grams of the powder (Zandu Pharmaceuticals, India) was boiled in 1000 ml of DDW till the volume
was reduced to one fourth of the original (250 ml). The
extract was cooled, centrifuged using a cold centrifuge
(Sorvall RC-5B, USA) and the supernatant was collected
and was concentrated by evaporating its liquid contents.
An approximate 26% yield of the extract was obtained.
Preparation of the drug and mode of administration

The required amount of triphala extract (TE) was dissolved in sterile double distilled water (DDW) and administered intraperitoneally.
Determination of acute drug toxicity

The acute toxicity of TE was determined according to

Prieur et al (1973) and Ghosh (1984). Briefly, the ani-

101

mals were allowed to fast by withdrawing the food and

water for 18 h. The fasted animals were divided into
several groups and each group of animals was injected
with various doses viz. 200, 220, 240, 260, 280, 300,
350, 400, 450, 500, 750 and 1000 mg/kg body weight
(b.wt.) of freshly prepared extract of TE intraperitoneally. Animals were provided with food and water
immediately after the drug administration. Mortality of
the animals was observed up to 14 days post-drug treatment. Acute LD50 of the extract was calculated using a
computer program for probit analysis.
Effect of TE on the radiationinduced mortality

The animals were divided into the following groups:

DDW + irradiation group
The animals of this group were administered with
0.01 ml/g b.wt. of sterile double distilled water (DDW)
intraperitoneally.
TE + irradiation group
The animals of this group were injected intraperitoneally with 5, 6.25, 10, 12.5, 20, 25, 40, 50 and 80
mg/kg b.wt. of TE consecutively for five days (Jagetia
and Aruna 1997).
Irradiation

One hour after the last administration of DDW or TE

on the 5th day, the prostrate and immobilized animals
(achieved by inserting cotton plugs in the restrainer) of
both the groups were whole-body exposed to 0 (shamirradiation) or 10 Gy of 60Co gamma radiation (Gammatron, Siemens, Germany) in a specially designed
well-ventilated acrylic box. A batch of 6 animals was
irradiated each time at a dose rate of 1.33 Gy/min at a
source to animal distance (midpoint) of 102 cm. The
animals were monitored daily up to 30 days post-irradiation for the development of symptoms of radiation
sickness, and mortality. The statistical significance between the treatments was determined by Z test.

Table 1. Acute toxicity of the extract of triphala on the survival of mice.

Triphala
mg/kg

Mortality on different days post drug treatment

1

200
220
240
260
280
300
350
400
500
750
1000

1
1
3
6
7
9
10
10

2
2
4
2
3
1

1
3
2

10

11

12

13

14

% Survivors

Survivors/Total

100
100
100
70
50
0
0
0
0
0
0

10/10
10/10
10/10
7/10
5/10
10/10
10/10
10/10
10/10
10/10
10/10

G. C. Jagetia et al.

1
4a
5b
7c
6c
4a
4a
3
0
0
2
2

2
1

2
1
3
1
2
2
1

1
2

1
1
1

1
1

1
1

1
1

1
1

9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30
8

5
2
1

2
2
1
1

Fig. 1. Effect of various doses of triphala extract (TE) on the

radiation-induced mortality in mice exposed to 10 Gy of -radiation. a) 10 day and b) 30 day survival.

P < a = 0.02; b = 0.01 and c = 0.001

2
2

1
1
1
3

0
5
6.25
10
12.5
20
25
40
50
80

2
5

6 7
5
3 4
2
1

Mortality on different post-irradiation days

Triphala
(mg/kg)

Table 2. Effect of various doses of triphala on the survival of mice exposed to 10 Gy of -irradiation.

No. of
Survivors

25
12
12
12
12
12
12
12
12
12

Total

102

 Results
Acute Toxicity

The administration of different doses of TE viz. 200,

220, and 240 mg/kg b.wt. did not induce any mortality
during the whole observation period. However, a further increase in the drug dose up to 260 mg/kg b.wt. resulted in a 30% reduction in the survival of mice. An
increase in the dose of TE up to 280 mg/kg b.wt.
caused a 50% reduction in the survival of mice. 100%
mortality was observed at 300 mg/kg and thereafter up
to a dose of 1000 mg/kg b.wt. of TE (Table 1).

The evaluation of the radioprotective effect of Triphala

Effect of TE on the radiationinduced mortality

The mice were treated with different batches of 0, 5,

6.25, 10, 12.5, 20, 25, 40, 50 and 80 mg/kg b.wt. of
triphala extract, consecutively for 5 days before wholebody exposure to 10 Gy gamma radiation or not were
monitored daily up to 30 days post-irradiation for the
development of symptoms of radiation sickness, and
mortality. The effect of different doses of TE on the radiation-induced mortality is shown in Table 2 and Fig 1
and the results are expressed as percent survival.
The animals of DDW+irradiation group exhibited
signs of radiation sickness within 24 days after exposure to 10 Gy of -radiation. The main symptoms included reduction in the food and water intake, irritability, epilation, weight loss, emaciation, lethargy, diarrhea, and ruffling of hairs. Facial edema was also observed in a few animals between one and two weeks
after exposure. During the second week after exposure
there were a few cases of animals, exhibiting paralysis
and difficulty in locomotion. The first mortality in this
group was observed on day 4 and 80% of the animals
died within 10 days after irradiation, while 96% of the
animals died within 30 days of irradiation resulting
only in 4% survival by day 30 post-irradiation.
The daily administration of 5, 6.25, 10, 12.5, 20, 25,
40, 50 and 80 mg/kg b.wt. TE for five consecutive days
did not cause any drug-induced mortality. The pretreatment of mice with various doses of TE either delayed
or reduced the severity of radiation sickness. The onset
of radiation-induced mortality was also delayed in
TE+irradiation group when compared with the
DDW+10 Gy irradiation group. The longest delay was
observed for 20 mg/kg TE, where the first mortality
was observed by day 10 post-irradiation (Table 2),
while a shortest delay was observed for 80 mg/kg,
where the first mortality occurred on day 3 post-irradiation.
Treatment of mice with various doses of TE also had
an ameliorating effect on the gastrointestinal tract as
evidenced by an increase in the 10 day survival of
mice, where an increase of 4.58 fold was observed for
12.5 mg/kg, 4.16 fold for 6.25, 10, and 20 mg/kg TE,
3.33 for 25 mg/kg and 2.5 fold for 5 and 40 mg/kg TE
(Fig. 1). Majority of the animals (80%) of DDW+ irradiation group, died within 10 days after irradiation,
while the TE pre-treatment increased the 10-day survival significantly. A lowest mortality was observed in
the animals treated with 12.5 mg/kg TE before irradiation and this decline in mortality was significant (p <
0.001). The other doses of TE also reduced the mortality, in comparison with DDW+ irradiation, however, a
significant elevation in the 10-day survival was observed only for 6.25, 10, 12.5 and 20 mg/kg TE (p <
0.001) treatment. 5 and 25 mg/kg of TE also protected

103

mice against the radiation-induced mortality, however,

the differences were statistically non-significant.
Analysis of thirty day survival revealed a drug dose
dependent increase in the survival of irradiated mice up
to a dose of 10 mg/kg in the TE+irradiation group,
where a highest survival of 58.33% was observed as
compared to the DDW+irradiation, where only 4% animals survived at the end of 30 days (Table 2). A further
increase in the drug dose to 12.5 mg/kg resulted in
8.33% reduction in the survival, when compared with
the 10 mg/kg TE. Increase in TE doses further, resulted
in a consistent decline in the animal survival reaching a
nadir at 50 mg and 80 mg/kg, where no survivors could
be reported at the end of 30 days (Fig 1). TE administration before irradiation increased the survival significantly for 5 to 25 mg/kg (p < 0.02 to 0.001). However,
the number of survivors was highest (58.33%) after
pretreatment of mice with 10 mg/kg of TE, and hence it
was considered the optimum dose for radioprotection.
The optimum radioprotective dose of 10 mg/kg TE was
found to be 1/28 of the LD50 dose (280 mg/kg b.wt.),
which was far below the LD50 dose.

 Discussion
Ayurveda (in Sanskrit Ayu = life and veda = knowledge), the Indian system of medicine, dating back to
5000 years has been an integral part of Indian culture
and materia medica. The Ayurveda, extensively uses the
plant-derived compound formulations for the treatment
of various ailments after a careful study into the type of
the disease (Sivarajan and Balachandra, 1996). Often
the drugs formulated are such that they have the desired
activity with the adequate potency and are devoid of untoward side effects. As it is observed that the desired activity is rarely present in adequate potency in a single
plant and it may also contain unwanted activities. Therefore, several plants with the common desired activities
and varied undesirable activities are selected so that the
final formulation will have a concentrated desired activity and the undesired activities will be absent or diluted.
Further, it is also observed that in such formulation, certain other compound may be of help in enhancing of the
potency of the active compounds resulting in an additive
or synergistic positive effect, which may be of immense
benefit to the patient (Kulkarni, 1997). Keeping this
Ayurveda philosophy in mind, triphala, an herbal
rasayans preparation, credited with diverse beneficial
properties like anti-aging, antimutagenic, anticancer, antibacterial, anti-viral, cardioprotective, hepatoprotective, anti-stress, cleanser of colon, gas distentioner, antidiabetic, antiparasitic, anti-diarrhea and antianemic
(Nadkarni, 1976; Mehta et al., 1993; Ahmad et al., 1998;
Phadke and Kulkarni, 1989; Niwa et al., 1995; Valsaraj

104

G. C. Jagetia et al.

et al., 1997) has been selected for the evaluation of its

radioprotective activity in mice.
The animals of the irradiated group (DDW + irradiation) exhibited signs of radiation sickness within 24
days after exposure to 10 Gy and the symptoms included reduction in the food and water intake, irritability,
epilation, weight loss, emaciation, lethargy, diarrhea,
and ruffling of hairs. The death of 80% of the animals
exposed to 10 Gy of radiation within 10 days is because
of functional failure of the gastrointestinal tract (Bond
et al., 1965; Uma Devi et al., 1999). The remaining 16%
animals died within the next 20 days exhibiting
hemopoietic syndrome and the charecteristic symptoms
like, irritability, epilation, weight loss, emaciation,
lethargy and ruffling of hairs (Bond et al., 1965; Uma
Devi et al., 1999). It is a well-established fact that ionizing radiation at cellular level can induce damage in the
biologically important macromolecules such as DNA,
proteins, lipids and carbohydrates in the various organs.
While some damage is expressed early others are expressed over a period of time depending upon the cell
kinetics and the radiation tolerance of the tissues. Like
in chemotherapy, the effect of whole body irradiation is
mainly felt by the highly proliferating germinal epithelium, gastrointestinal epithelium and the bone marrow
progenitor cells. Of these the germinal epithelium does
not have a life supporting function to the exposed individual, while the gastrointestinal epithelium and the
bone marrow progenitor cells are crucial for sustaining
of life and any damage to these cells will impair the normal physiological processes drastically. The gastrointestinal epithelium is less sensitive than the bone marrow progenitor cells but as the cell transit time is quick,
it is expressed earlier than the hemopoetic syndrome
(Bond et al., 1965). In mice death within 10 days postirradiation is due to the gastrointestinal damage (Uma
Devi et al., 1999). The bone marrow stem cells are more
sensitive to radiation damage than the intestinal crypt
but, the peripheral blood cells have a longer transit time
than the intestinal cells and hence the gastrointestinal
syndrome appears earlier than the bone marrow syndrome and in mice death due to irradiation from 11 to
30 days is due to the hemopoetic damage inflicted by
radiation (Bond et al., 1965; Uma Devi et al., 1999).
The pretreatment of mice with different doses of TE
resulted in a dose dependent reduction in the radiationinduced mortality up to 10 mg/kg and a further increase
in the drug dose resulted in the decline in the animal
survival when compared with the 10 mg/kg TE. The
earlier studies on radioprotection have shown that an
agent in test (for radioprotective action) acts only at a
particular dose range and above which it may not be
protective and some times can even be toxic (Thomson, 1962; Yuhas and Storrer, 1969). The active principle of Plumbago rosea, the plumbagin at pico to femto

gram range has been reported to stimulate the granulocytes in vitro, while at higher doses it had immunosuppressive activity (Wagner et al., 1988). The reason may
be that after a particular concentration, a compound instead of being an anti-oxidant may act like a pro-oxidant inducing toxic symptoms resulting in the death.
This is the reason that TE has optimum protection at 10
mg/kg and the higher doses result in the decline in the
protective action of TE. The TE pretreatment provided
protection against the radiation sickness and mitigated
the animal sufferings. Reports regarding the use of TE
to protect against the radiation damage are unavailable,
as this is probably the first report regarding the radioprotective action of TE. However, certain other herbal
preparations like Liv. 52, and abana have been reported
to protect the mice against the radiation-induced sickness, mortality, dermatitis, spleen injury (Saini et al.,
1984 a, b) and radiation-induced chromosome damage
(Jagetia and Ganapathi, 1989, 1991; Jagetia and Aruna,
1997). The brahmarasayana, narasimharasayana, ashwagandha-rasayana, and amrithaprasham, a group of
herbal preparations used to improve the general health
and debility, have been reported to reduce the radiation-induced lipid peroxidation in the liver, and leucopenia in mice (Kumar et al., 1996).
The pattern of survival in TE+irradiation group was
similar to that of the irradiated control group except
that the mortality was delayed. This clearly indicates
the effectiveness of TE in arresting GI death, where the
number of survivors for 5, 6.25, 10, 12.5, 20, 25 and 40
mg/kg was higher than that of the irradiated control.
The reduction in GI death may be due to the protection
of intestinal epithelium, which would have allowed
proper absorption of the nutrients. Triphala has been
used as laxative to support the bodys vitality in man
and it even stops diarrhea. Our findings support the
contention that triphala may protect the gastrointestinal
tract epithelium against the toxic insult of radiation,
protecting against the GI death in this study. It has been
reported that, Terminalia chebula, an important constituent of Triphala, mitigated the cysteamine-induced
duodenal ulcers in rats by increasing the beta-glucuronidase activity in the Brunners glands (Nadar and
Pillai, 1989) and protected the epithelial cells against
the cytopathic effects caused by influenza A virus
(Badmaev et al., 2000). Another herbal drug Liv. 52
has been reported to protect the intestinal epithelium
against the radiation-induced damage (Saxena and
Goyal, 1998).
The pretreatment of mice with TE significantly reduced the bone marrow deaths in the TE+irradiation
group, especially at a dose of 5 to 25 mg/kg, where a
significant elevation in the survival has been observed.
This increase in the 30 day survival may be owing to
the protection afforded by TE to the bone marrow stem

The evaluation of the radioprotective effect of Triphala

cell compartment, which continued to supply the requisite number of cells in the survivors. The bone marrow
cells have been reported to be protected against the radiation-induced damage by various other plant formulations (Saini et al., 1984a, b; Jagetia and Ganapathi,
1989, 1991; Jagetia and Aruna, 1997; Kumar et al.,
1996). Further, triphala, and its constituents are reported to possess antimicrobial activity (Mehta et al., 1993;
Dutta et al., 1998; Ahmad et al., 1998; Phadke and
Kulkarni, 1989; Valsaraj et al., 1997), which would
have also been responsible for the radioprotective action of TE. One of the constituents of triphala, the Phyllanthus emblica, has been found to be immunomodulator (Suresh and Vasudevan, 1994; Rege et al., 1999)
and this would have increased the bodys defence system by increasing the immunity. Further the antimicrobial action of triphala would have prevented the localization of the pathogenic microbes in the GI tract and
bacterial infection, resulting in the observed radioprotection.
TE is mainly composed of Terminalia chebula, Phyllanthus emblica and Terminalia bellerica in equal proportions and each plant has been utilized to treat various
ailments and diseases in the Ayurvedic system of
medicine. Terminalia chebula is a commonly advocated
agent in Ayurveda for improving gastrointestinal motility (Tamhane et al., 1997). The water and chloroform
extracts of it have been shown to protect against the
sodium azide and 4-nitro-o-phenylenediamine induced
mutagenesis (Grover and Bala, 1992). Recently, it has
also been reported to possess antioxidant activity and
prevent the TPA-induced DNA breaks in human white
cells (Naiwu et al., 1992). Similarly, Terminalia bellerica has been found to contain anti-HIV-1, antimalarial,
and antifungal activity (Valsaraj et al., 1997). The alcoholic extract of this plant reduced the serum GOT, GPT
and LDH activity, caused a significant reduction in fatty
acid levels, and protected against the myocardial necrosis (Tariq et al., 1977). Phyllanthus embelica has also
been found to be rich in ascorbic acid contents and
ascorbic acid has already been reported to reduce the radiation-induced sickness and mortality (Redpath et al.,
1982) and to protect mice bone marrow cells against the
radiation-induced chromosome damage (Sarma and
Kesavan, 1993). In addition to ascorbic acid, the components of triphala, Phyllanthus emblica, Terminalia
chebula and Terminalia bellerica also contain ellagic
acid, which has been reported to decrease the bone marrow micronuclei formation in the mice (Thresiamma et
al., 1996). Phyllanthus emblica has also been reported
to contain flavonoids (Jose and Kuttan, 1995), a class of
compounds reported to be possess antioxidant and free
radical scavenging activities (Tanaka, 1994; Uddin and
Ahmad, 1995; Korina and Afanasev, 1997; Uma Devi et
al., 2000). Certain flavanoids have been found to pro-

105

tect against the radiation-induced DNA damage (Shimoi et al., 1994, 1996, 1997; Uma Devi et al., 1998) and
mortality (Uma Devi et al., 1999). The aqueous, acetone and chloroform extracts of Emblica officinale
found to have antimutagenic effect (Grover and Kaur,
1989).
The exact mechanism of action of TE is not known,
however, it may scavenge free radicals produced by radiation and thus reduce the radiation-induced damage
to the cellular DNA. The presence of ascorbic acid and
the flavonoids like quercetin may be responsible for
this action as these compounds are reported to protect
the DNA from radiation-induced micronuclei in mice
(Sarma and Kesavan, 1993; Shimoi et al., 1997). While
testing NO (nitric oxide) scavenging activity of several
agents, triphala was found to scavenge the nitric oxide
production in vitro (unpublished data). The aqueous
extract of one of the constituents of TE, Phyllanthus
emblica has been reported to be a potent inhibitor of
lipid peroxidation formation, and scavenger of hydroxyl and superoxide radicals in vitro (Jose and Kuttan,
1995). Photochemical studies have shown that Terminalia bellerica contains bellericanin, ellagic acid, gallic acid, chebulagic acid, ethyl gallate and -sitosterol.
Terminalia chebula has been found to contain chebulin, terchebin, chebulagic acid, chebulinic acid, corilagin, ellagitannin, ellagic acid, gallic acid, -D-glcogallin, and terchebin. The Phyllanthus emblica has
been reported to be a rich source of vitamin C and also
contains terchebin, corilagin, tannins, ellagic acid,
phyllembic acid, gallic acid and flavonoids in different
proportions depending on the season, type of climate
and the plant processing (Chemexcil, 1992; Satyavati
et al., 1987; Wealth of India 1952, 1976; Rastogi and
Mehrotra, 1990; Jose and Kuttan, 1995). Most of these
compounds have been reported to possess antioxidant
and free radical scavenging activities (Tanaka, 1994;
Uddin and Ahmad, 1995; Korina and Afanasev, 1997)
and increase the antioxidant enzymes (Kong Ah-Ng et
al., 2000). The presence of various antioxidant compounds in triphala might have been responsible for the
observed radioprotection by scavenging of free radicals generated by radiation exposure. Alternatively
triphala might have increased the intracellular level of
GSH, and stimulated the immune systems which could
have provided protection against the radiation-induced
mortality.

 Conclusions
From our study it is clear that TE, a plant based formulation provided protection against the radiation-induced sickness and mortality and the optimum protective dose of 10 mg/kg i.p. is far below the LD50 (280

106

G. C. Jagetia et al.

mg/kg) dose. The exact mechanism of action of TE is

not known, however, it may scavenge free radicals produced by radiation and thus inhibit radiation-induced
damage to the cellular DNA. We have observed scavenging of NO (nitric oxide) radicals in vitro by TE (unpublished data) and this testifies to our belief that one
of the mechanisms of radioprotection by triphala may
be owing to the scavenging of free radicals generated
by radiation exposure. Alternatively, it may also increase GSH levels and may reduce the radiation-induced lipid peroxidation. Since significant protection
is obtained at a very low non-toxic dose the extract
may have an advantage over the known radioprotectors
available so far. Studies are planned to explore the applicability of triphala in cancer treatment by looking
for the preferential protection to the normal tissues and
its clinical applicability for cancer cure in the fractionation regime. Since this formulation has been in use in
India for the last 5000 years, the clinical acceptability
shall not be a problem.

 Acknowledgements
We thank Prof. M. S. Vidyasagar, and Dr. J. Velumurugan, Department of Radiotherapy and Oncology, Kasturba Medical College, Manipal, India for providing
the necessary irradiation facilities and help in radiation
dosimetry.

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 Address
Dr. Ganesh Chandra Jagetia, Department of Radiobiology, Kasturba Medical College, Manipal 576 119,
India.
Tel.: ++091-8252-71201 to 71300 ext. 22814;
Fax: ++091-8252-70062/71927;
e-mail: gc.jagetia@kmc.edu