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Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 1

1. Introduction
Immunomodulation is one of the most rapidly developing areas of medical science & disease
prevention research and has great promises with regard to the prevention and treatment of a
wide range of disorders such as the inflammatory diseases of skin, respiratory disorder and
various infectious diseases are disorders caused by organisms such as bacteria, viruses, fungi
or parasites. And worlds 25.9% of the death occurred due to the infectious diseases. 1 In
addition, infectious diseases are now primarily considered immunological disorders.
Immunomodulators are natural or synthetic substances that help regulate or normalize the
immune system. Immunomodulators correct immune systems that are imbalance. This
elaborate defence system can keep health problems ranging from HIV/AIDS to the common
cold at bay. Research on immunomodulation by natural products or synthetic derivatives is of
key interest for immunomodulation therapy for a number of reasons. Many plant remedies
well-known in traditional medicine or refined natural products in clinical use by directly
affecting the pathogen. At least part of their effect is indirect, by stimulating natural and
adaptive defence mechanisms of the host. These findings have now given many empirical
therapies a rationale, scientific basis and thereby a means for intelligent improvement. In
discovering the molecular mechanisms by which known remedies exert their effects, chosen
elements further down the chain of command might be synthesized and applied directly for
more rapid and selective cure, omitting unwanted side effects. The direct use of recombinant
cytokines, often in combination with antibiotics, is one consequence of this rationale.
As herbal immunodulatory drugs has the potential to achieve the greater efficacious drugs for
the prevention of infectious diseases.
Ayurveda antiquity developed certain dietary and therapeutic measures to arrest/delay
ageing and to rejuvenate whole functional dynamics of the body system. This revitalisation and
rejuvenation is known as the rasayan chikitsa (rejuvenation therapy) which, in the current
context, can be equated to immunomodulation or adaptogenic activity. Traditionally, rasayana
drugs are used against a plethora of seemingly diverse disorders with no pathophysiological
connections according to modern medicine. Although this group of plants generally possesses
strong immunomodulatory activity, only some have been investigated in detail. In this study,
apart from an insight into the role of Ayurveda and rasayana in the modulation of the immune
system, immunomodulatory activities of other medicinal drugs, along with some compounds
isolated from plants are reviewed. Ayurveda physicians believe in preventive therapy rather
than curative and the rasayanas are the disease-preventive agents of Ayurveda.2

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Semecarpus anacardium (SA) Linn. (Family Anacardiaceae), commonly called as
Bhallatak, It is well-known for its Rasayana properties in Ayurveda. Immunomodulation using
SA provide an alternative to conventional chemotherapy for a variety of diseases, especially
when host defence mechanism has to be activated under the conditions of impaired immune
response or when a selective immunosuppression is desired in situations such as autoimmune
disorders.3 It appears that the normal way by which the immune system works is through its
own modulation by factors usually synthesised by the immune cells.4
SA has been shown; the nut oil from Semecarpus anacardium (SA) was shown to be
cytotoxic to human leukaemic cell lines.5 The ethyl acetate extract contains a biflavonoid
known as tetrahydroamentoflavone (THA) that inhibited the enzyme cyclooxygenase-2 (COX2).6 The alcoholic extract of dry nuts had anti-fungal activity while of nut shells were shown to
prevent lipid peroxidation.7 The nut milk extract is effective in restoring the fragility of
lysosomal membranes in aflatoxin-induced hepatocellular carcinoma.8 The immune response
requires timely interplay of multiple cell types within specific microenvironments to maintain
immune homeostasis. The selectivity and flexibility that is necessary to regulate cell traffic
under homeostatic and diseased conditions are provided by the differential distribution and
regulated expression of cytokines and their receptors. As a consequence, cytokines are
responsible for the development of phenotypes and are, therefore, logical targets for therapeutic
immune modulation.9
1.1 Classification of immunomodulation
In clinical perspectives, immunomodulators can be classified into three categories.
Immunoadjuvants are used for enhancing the efficacy of vaccines and therefore could be
considered to be specific immune stimulants. Adjuvants such as monophosphoryl lipid A,
immunostimulating complex (ISCOM)10
Immunostimulants are compounds leading predominantly to a non-specific stimulation of
the immune system and are also called mitogens. These agents, generally, interact not just
with one but also with other types of immune competent cells, because of the close link between
the non-specific and specific immune system. This is one handicap in developing effective
immunostimulating agents without any side-effects, since in some cases immunostimulants
may also stimulate T-suppressor cells, and thereby reduce the immune resistance. The terms
immunomodulators or immunoregulators therefore very often seem to be more appropriate.11
Immunosuppressants are agents that could be used for control of pathological immune
response in autoimmune diseases, graft rejection, graftversus- host disease, hypersensitivity

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immune reaction (immediate or delayed type) and immune pathology associated with
infections.12

1.2 Immunomodulators in Ayurveda


A significant part of Ayurvedic therapeutics is preventive in nature. The capacity of the body
to resist disease, is stimulated. The rasayanas of Ayurveda are believed to be
immunomodulators in modern terminology. Ayurveda considers that an individual, when
advanced in age, accumulates wastes and toxic substances in his/her cellular system, which
disrupt the normal metabolism, leading to loss of immunity, causing disorders or diseases and
finally hastening the ageing process. Ayurvedic experts have developed various methods to
detoxify the tissues and rejuvenate the body by a special treatment regimen called rasayana
chikitsa. The immune system is known to be involved in the aetiology of many diseases as well
as in the pathophysiological mechanisms.
Ayurveda emphasises the promotion of health, a concept of strengthening host defences
against different, so rasayana drugs are particularly recommended for the treatment of immune
disorders. The development of agents capable of moving the patients immune system from a
state of deficiency to one of more normal function would be likely to have a significant impact
on disease. Such agents would not be a cure but would control the manifestation and course of
disease some plants and their constituent compounds are claimed to induce para immunity, the
non-specific immunomodulation of macrophages, granulocytes, natural killer cells and
lymphocytes and complement functions13. Many studies have been undertaken to provide
scientific support for the use of rasayana drugs as immunomodulators and adaptogens.
Rasayana drugs have been reported to treat generalised weakness

14

and to afford protection

from cyclophosphamide-induced leukopenia.15


1.3 Concept of 'Vyadhirodhak Chamatav'.
There is a difference in the concept of bodys resistance to disease in traditional Indian
system of medicine, According to Ayurvedic theory a harmonious balance between
three humors of the body viz. Vayu', Pitta and Kaph is needed for positive health; imbalance
of these may cause disease(s). A significant part of Ayurvedic therapeutics is preventive in
nature. It aims to promote positive health so that individuals do not suffer from disease. This
is the concept of "Vyadhirodhak chamatav". ie capacity of the body to resist disease.
Obviously, the immune system. As recognised in modern biology which provides protection
against microbes, should be a part of it. An entire section of the Materia Medica of Ayurveda

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 4


termed Rasaynas' is devoted to enhancement of body's resistance. Interestingly the
prescribed this section include not only procedures under drugs ('Aushadhi') but also "Aachar"
(daily routine including exercise), "Aahar" (diet and nutrition) and "Vyavhar (mental attitude
and discipline) which are equally important in achieving the desired goal. In comparison to this
concept of Rasaynas' in Ayurveda.16

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2. Literature Review
2.1 Classical Text
2.1.1 Charak samhita:
According to charak samhita, chikitsasthanam, the sneha of bhallatak is taken into one vessel
along with godugdha mulethi is & cooked with the oil. The same process repeated for 100
times. The uses are generally similar to that of bhallatakksoudhra

2.1.2 Susruta samhita:


In suhruta Samhita, properties and uses of bhallatak oil mentioned Bhallatak oil uses are is
bakarak, balya

2.1.3 Shankar nighantu:


In Shankar nighantu, different synonyms like bhallatak ,bhilawa, bhela, marking
nut,malaccabean, and properties like astringent, hot ,sukarjanan, madhur , and light it is used
in vatalkapha, udarroga, kustha, babascer, sangrahniya, gulam jawara, agniga, mandya,
krimirog and varann The friuts of ballatak sweet, when ripe, light,astringent, snigdha, tikana,
hot, chedan, bhedan and appetizer. The bark of fruit is sweet, light, astringent, and it is used in
deepan, pachan, uderrog, swelling and fever.
Shodan of ballatak: Keep the ripe fruits of ballatak in water, fruits that are sink into the water,
that take for the shodahan with the same quantity of water, then rubbed on the brick powder.

2.1.4 Bhav prakash nighantu:


It is a 22-40 feet long tree bark.1 inch thick and grayish in colour. Flowers are inflorescence
yellowish in colour. Fruits are l inch long, black in colour. Different properties, chemical
composition and there therapeutics uses and shodahan of bhallatak mentioned in Bhavpraksh
nighantu.

2.1.5 Nighantu Adarsh:


Bhallatak, aruskar, agnimukh, bhilawa, biladur, veervariksha. It is astringent, sweet, hot, bitter
and kaphavata nashka. It is a medium size tree. Found in Assam, madhyabharat and
dakshinabharat. The leafs are long, ovate, Flowers are yellow in colour.The fruits are heart in
shape, black and white in colour when unripe'

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2.1.6 Raj nighantu:


Bhallatak, agni, dahan, patan, aruskar, anal, krimighan tailbeej, vaatari, sufoot bijak,
prithakbeej, dhanuurbeej, beejeepadak,etc. Sixteen synonyms are given. It is katu, tikta, kasaya
in rasa. It is used in vata disorders, udarvikar, anaah and parmeha. The fruits of bhallatak are
madhur rasa in and ushna. It is used in kapha, sarram, swasarog, annah, vibandsool, udarrog
and kriminashak

2.1.7 Dhanvantari nighantu:


Aruskar, dahan, tapan, agnik, bhallatak, agnimukha, virtaru and dhanu. It is katu tikta, madhur,
ushna virya, krimighan and vata, pitta nashak

2.1.8 Priya nighantu:


Bhallatak is sharp like bhala weapon. It cause irritation when touch the fruit without precaution.
It is used in kusta, arsha, kapharog, raktadustijanya vikar, agnimandya, krimiroga.
2.1.9 Dravyaguna Vijnana:
Botanical description: A moderated size deciduous tree, exudation a dark juice. Young
branches inflorescence petiole and under side of leafs pubescent. Leaves- oblong, ovate
rounded at apex, cartilaginous at margin, very coriaceous. Flowers- fasciculate, arranged in
erect, compound greenish yellow colour. Fruits drupes, obliquely oval or long, smooth, shining,
purplish-black when ripe, cup orange. Flowering round the year, mostly during May-June,
fruits ripe from November to February.17
2.1.10 Ayurvedic pharmacology:
Class: Kushthaghna, Deepaniya, Mootrasangrahaniya, Nyagrodhadi, Mustadi
Kula: Amra kula
Family: Anacardiacee (Ana-like, Cardium-heart)
Method of purification: Take out the bhallataka fruit along with the stalk and keep it in the
powder of bricks for period of one week. Clean and wash thouroughly by a rubbing it, then
boil it with milk. This purifies bhallataka impure bhallataka act as a toxin, hence it should be
used without purifies. Small children, pregnant not women, older people, persons having
pittaprakruti, patients having tendency to bleed, and those who are allergic to Bhallatak should

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not take bhallataka. It is important for a vaidya to know the contraindication rather than the
indication of bhallataka18
2.1.11 Ayurvedic pharmacopeia of India:
Bhallataka consists of mature fruit of Semecarpus anacardium Linn. (Fam.Anacardiaceae), a
medium sized tree found in moist deciduous forests all over the country.
Synonyms
Sanskrit

: Bhallatakam

Assamese

: Bhelaguti

Bengali

: Bhela

English

: Marking Nut

Gujrati

: Bhilam

Hindi

: Bhilawa

Kannada

: Bhallataka

Malayalam

: Chera

Marathi

: Bibba

Oriya

: Bhollataki, Bholai

Punjabi

: Bhilawa

Tamil

: Tatamkottai, Scramkotati

Telugu

: Nallajidi, Nallajidiginga

Urdu

: Baladur, Bhilavan

Description
Macroscopic: Fruit laterally flattened, drupaceous, dark brown, nut 2.5-3 cm long, obliquely
ovoid, smooth, shining with residual receptacle.
Microscopic:
Fruit - Pericarp differentiated into epicarp, mesocarp and endocarp; in longitudinal section
pericarp shows outer epicarp consisting of single layer of epidermal cells which are elongated
radially and lignified, characteristic glands found in pericarp which exude oil globules and arise
as small protuberances in epicarp and due to pressure exerted by cells of mesocarp, some of
epidermal cells and cuticle rupture and oil globules exude from oil glands; mesocarp a very
broad zone, 30-40 layers thick, composed mostly of parenchymatous cells having lysigenous
cavities and fibro-vascular bundles, below epidermis a few outer cells of parenchyma smaller
as compared to rest; rosette crystals of calcium oxalate found scattered in parenchymatous

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cells, some cells get dissolved and form lysigenous cavities which increase in size with maturity
of fruit, cavities do not have any special lining and contain an acrid and irritant yellowish oily
secretion; endocarp consists of two distinct layers, innermost prismatic, very much elongated
radial walls, being highly thickened, outer layer shorter and thinner than prismatic layer but
cells similar to the former; number of mesocarp parenchyma contain rosette crystals of calcium
oxalate and oil drops in oil glands; lysigenous cavities of mesocarp contain oily vesicating
substance, insoluble in water and soluble in alcohol, ether, chloroform. Powder - Dark-brown;
shows rosette crystals of calcium oxalate and oil globules.
Identity, Purity and Strength
Foreign matter

: Not more than 1 %

Total Ash

: Not more than 4 %

Acid-insoluble ash

: Not more than 0.5 %

Alcohol-soluble extractive : Not less than 11 %


Water-soluble extractive

: Not less than 5 %

Constituents: A Tarry Oil containing Anacardic Acid, Non-Volatile Alcohol (Cardol).


Properties and action
Rasa : Madhura, Katu, Tikta, Kasaya
Guna : Laghu, Snigdha,Tiksna
Virya : Usna
Vipaka : Madhura
Karma : Dipana, Kaphahara, Pacana, Vatahara, Chedi, Bhedi, Medhya
Important formulations: Amrta Bhallataka Leha, Bhallataka
Rasayana, Bhallatakadi Modaka
Therapeutic uses: Arsa, Anaha, Grahani, Gulma, Krimi, Kustha.
Dose: 1.2 g. of the drug in Ksirapaka form.19

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2.2 Reported pharmacological activities of Semecarpus anacardium.


Table 1: Reported pharmacological activities of SA
S.
No
.

1
2

Extract/
Formula
tion

Pharmacological
activity reported

Nut
Breast cancer
extract
Seeds
COX inhibitors
(Biflavan
oids)

Nut milk
extract

Antidiabetic

Nut
extract
Nut milk
extract
Nut
extract

Antiarthritic

5
6

Antiarthritic
Immunomodulato
ry and
Antiarthritic
Antiarthritic

In vivo
models
(Humans/An
imals)

Male spargue
Dawley rats
(Rat paw
edema assay)
Male albino
wistar rats

400
mg/mL
100 g/kg

20

22

21

150 mg/kg

24

1g/mL

25

Male albino
wistar rats
(Freunds
adjuvant
induced
arthritis)
Male albino
wistar rats

150 mg/kg

26

150 and
300 mg/kg

27

DPPH and
ABTS assay

100
g/mL
(DPPH
assay) and
0.01 to 0.5
mg/mL (
ABTS
assay)
200 mg/kg

28

Chlorofo
rm
extract
of SA
Ethanol,
acetone
and
aqueous
extract

Aphrodisiac

Antioxidant

Nut milk
extract

Antidiabetic (type
II)

Male spargue
Dawley rats
(Streptozotoc
in induced

10

Ref

Male albino
wistar rats
Male albino
wistar rats
Humans

Nut milk
extract

Dose

300 mg/kg
b.wt. (For
21 days)
150 mg/kg

In vitro
models (cell
lines/
chemical or
microbial
assay)
T47D cell
line
-

23

29

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diabetes
mellitus)
Male albino
wistar rats

11

Fruit
extract

Hepatoprotective

12

Methano
lic
extract

Anti fungal

13

Petroleu
m ether
nut
extract

Antibacterial

250 and
500
mg/kg
6.25,12.5,
25, 37.5,
50 and
62.5
g/mL)

Fungal
strains
( Fusarium
oxysporum,
Rhizctonia
solanii,
Alternaria
spp., and
Sclerotium
rolfsii)
Microorganis
m
(E.Coli,Bacill 150 l
us
subtilis,Micro
coccus
luteus,Klebsi
ella
pneumonia,
Streptococcu
s
aureus,Proteu
s vulgaris,
Salmonella
typhi)

30

40

40

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14

15

16

Aqueous
and
organic
extract

Antimicrobial

Ayurvedic formulation.
Amrut
General tonic and
Bhallata vitalizer
kavaleha
(Electuar
y)
Bhallata Neuralgia and
kasava
asthma
(Wine)

Microorganis
m
(Staphylococ
cus aureus,
Shigella
flexneri,
Bacillus
licheniformis,
Vibiro
cholera,Pseud
omonas
aeruginosa,
Streptococcu
s aureus,
Bacillus
brevis.)

10,50,100
mg/ml
10mg
G/ml

31

Humans

1 to 2
teaspoonfu
l for 2
times

32

Humans

2 to 4
teaspoonfu
l for 2
times
2 pills
(500 mg
pill) for 2
times
2 pills
(250 mg
pill) for 3
times
250 mg
for 3 times

41

1 to 2 gm
for 2 times

41

17

Suran
vatak
(Pills)

Piles and
anorectal diseases

Humans

18

Sanjeeva
ni Vati
(Pills)

Dysentry and
diarrhea

Humans

19

Bhallata
k Parpati
(Powder)
Narsimh
a
Choorna
(Powder)

Rheumatic
diseases

Humans

General
restorative

Humans

20

2.3 Various Antidotes of SA Toxicity

41

41

41

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Cows milk, Cows Urine is recommended as the antidote for bhallataka blisters. Neem is used
as topical Antidotes.33
2.4 Drug interaction of SA
2.4.1 Manodeep Chakraborty et al., 2010 have reported Interaction of Semecarpus
anacardium L. with propranolol against isoproterenol induced myocardial damage in
rats
With a view to evaluate the cardio protective effect of ethanolic extract of S. anacardium nut
and the possible interaction with propranolol against isoproterenol induced myocardial damage
in rats, female Sprague-Dawley rats were pre-treated with propranolol (10 mg/kg for 7 days),
low and high doses of S. anacardium (100 and 500 mg/kg for 21 days) and their combination
orally and subsequently subjected to isoproterenol administration (150 mg/kg, sc) for two
consecutive days. The influence of prophylactic treatment was analysed by quantification of
biomarkers and antioxidants, electrocardiographic parameters and histopathological
observations. The activities of lactate dehydrogenase and creatinine phosphokinase-MB were
reduced in serum and raised in heart tissue with concurrent elevation in superoxide dismutase
and catalase activities as well as reduction in thiobarbituric acid reactive species levels
significantly in all treated groups compared to isoproterenol group. Similarly,
electrocardiographic changes were restored to normalcy in all treated groups. To conclude,
combination of high dose of S. anacardium with propranolol was found to be most effective in
alleviating the abnormal conditions induced by isoproterenol.
This study was to elucidate the role of S. anacardium nut extract during
myocardial dysfunction and metabolic derangement induced by isoproterenol in rat heart and
also to explore its pharmacodynamics interaction with conventional cardio protective drug.
Propranolol. The results revealed the beneficial role of S. anacardium when treated
concurrently with propranolol in conditions of anticipated cardiac injury.
Results it may be concluded that the SANE both at low (100 mg/kg) and high doses (500
mg/kg) possess cardio protective efficacy when given prophylactically in experimental
animals. Moreover, combined therapy of SANE and PRO demonstrated synergistic cardio
protective potential than when they were used alone. The combination of high dose of SANE
and PRO was found to have best effect. However, combined therapy of SANE and PRO must

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be further refined for adjustment of doses as higher doses of SANE and PRO may pose
negative implication due to excessive pharmacological effects.

2.5 Adverse effects of bhallataka:


Impure or pure bhallataka if taken in excess dose, cause pruritus and burning sensation of anus
and tip of the penis, excessive perspiration, thirst and reduced red dusky urination, in such
condition, one should or stop taking bhallataka and should start pitta alleviating drug like
doorvadigana, sarivadigana. In condition like burning sensation or pruritus and swelling of
skin, sesamum oil, coconut oil, resin ointment should be applied locally: In 3-4 ghee or days
the above symptoms subside. The above should be kept handy before starting Bhallatak
treatment.34

2.6 Safety profile:

The drug should be used with care, preferably under the direction of a qualified practioner,
since the Anacardic acid are allergenic. The maximum tolerated dose of a 50% alcoholic extract
of the fruit when given interaperitoneally to mice was found to be 250/mg/kg body weight.35

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3. Phytochemical profile of Semecarpus Anacardium.
SA can be greatly aided by the isolation of its active principle and determination of structure
and function relationship. Based on this principle a lot of phyto pharmaceuticals from different
parts of S. anacardium have been isolated. Phytochemical examination revealed 3.85% of total
ash, 0.33 % of acid insoluble ash, 11.27 % alcohol soluble extractive, 11.84% water soluble In
alternative medicine, medicinal plant preparations have found wide spread use particularly in
the case of diseases not amenable to treatment by modern methods. A variety of nut extract
preparations from S. anacardium are effective against many diseases viz. arthiritis, tumours,
infections etc.36 Various anacardoflavanone have been isolated from the nut shells and
characterized isolated one more biflavonoid namely tetrahydrorobustaflavone from the
defatted nuts of the S. anacardium and structure characterized. The leaves of the S. anacardium
found to contain amentoflavone as the sole compound. 37The corrosive juice from the pericarp
of the fruit found to contain catechol, fixed oil and anacardol (C18H13O3.COOH) to which the
corrosive properties of the juice are due to two phenolic acids C16H15O3.COOH and
C14H13O3.COOH. 38 From the seeds of S. anacardium, a new phenolic glucoside, anacardoside,
was isolated, and its structure and configuration were elucidated by a combination of NMR
techniques as-l-O- -D-glucopyranosyl- (1 - 6) - -D-glucopyranosyloxy-3- hydroxy-5methylbenzene.39

Table 2: Qualitative Phytochemical Analysis of Various Extracts of Semecarpus


anacardium Leaves.

S. No.
1.

Name of
Procedure
Alkaloids

the

Test Observation Soxhlet Extracts of Leaves


A B C

D E

F G

H I

J K L

M N

Dragendo
rffs Test

Orange Red ppt.

- -

- -

- -

ii

Mayers
Test

- -

- -

- -

+ -

iii

Hagers
Test

Whitish
Yellow or
Cream
coloured
ppt.
Yellow
coloured
ppt.

- -

- -

- -

+ -

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iv

Wagners
Test
Saponins

+ +

D E

F G

Foam
+ - persists for
10mins.
Carbohydrates
A B C

Molisch
Test

ii

Fehlings
Test
Benedict
s Test
Glycosides

2.
i

3.

iii
4.
i
ii

Reddish
Brown ppt.

+ +

H I

J K L

M N

- -

- -

+ -

D E

F G

H I

J K L

M N

- -

- -

- -

- -

- -

- -

+ -

- -

- -

- -

+ -

D E

F G

H I

J K L

M N

Pink to Red + - +
color.
Yellow to + + Orange
color.
A B C

+ -

- +

+ -

- +

+ -

D E

F G

H I

J K L

M N

A B C

Foam
Test

Legals
Test
Baljet
Test

Purple or reddish
violet color.
Brick Red ppt.
Red ppt.
-

A B C

5.

Tannins

Lead
acetate
Test
Ferric
chloride
Test
Potassium
dichromat
e Test
Gelatin
Test
Potassium
ferric
cyanide
Test
Flavonoids

White ppt.

Shinodas
Test
Alkaline
Reagent
NaOH
Test
H2SO4
Test

Cherry Red color.


Intense
Yellow
color.
Yellow
Orange
color.

ii

iii

iv
v

6.
i
ii

iii

- +

- +

- -

- -

- -

Dark Blue or Greenish


Black.
Yellow
color ppt.

- -

- -

- -

+ +

- +

+ -

White ppt.

- +

- -

+ -

+ -

red -

- -

- -

- -

D E

F G

H I

J K L

M N

- -

- -

- -

+ -

- +

+ -

+ +

+ +

- +

+ +

Deep
color.

A B C

or -

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iv

Lead
acetate
Test
Steroids

Yellow
color ppt.

Salkowsk
i Test

ii

Liberman
n
burchard
Test

8.

Phenols

Ferric
chloride
Test
Proteins

7.

9.
i

ii
iii
10.
i
11.
i

ii

iii

+ -

- -

D E

F G

Bluish red - + to
cherry
color
in
Chloroform
layer
&
Green
in
acid layer.
Brown ring - + at junction
& green or
deep
red
upper layer.
A B C

Bluish
Black color.

A B C

- -

A B C

Biuret
Test

+ +

H I

J K L

M N

- -

- +

- +

+ +

D E

F G

H I

J K L

M N

- -

- -

D E

F G

H I

J K L

M N

- -

Pinkish or Purple
violet color.
Ninhydrin Blue color. Test
Xanthopr Orange
oteic Test color.
Monosaccharide
A

- -

- -

- -

- -

- -

- -

- +

- -

+ -

+ -

B C

D E

F G

H I

J K L

M N

Barfoeds Red ppt.


Test
Hexose Sugars

- -

- +

- -

A B C

D E

F G

H I

J K L

M N

Selwinoff
s Test for
ketohexos
e
like
fructose
Tollens
phloroglu
cinol Test
for
galactose
Cobalt
chloride
Test

- -

- -

+ -

Yellow to +
Red color.

+ +

+ +

+ +

+ -

Upper layer +
Greenish
blue
&

+ +

+ +

+ +

+ +

Red color.

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 17


Lower
Purplish.
12.

Diterpenes

A B C

D E

F G

H I

J K L

M N

Copper
Emerald
acetate
Green
Test
color.
Nonreducing
Polsaccharides [Starch]
Iodine
Blue color.
Test
Tannic
ppt
acid Test formation.
Mucilages & Gums

- +

+ +

A B C

D E

F G

H I

J K L

M N

- -

- -

- -

- -

- -

- -

A B C

D E

F G

H I

J K L

M N

Rutheniu Pink color.


m
Red
Test

- +

+ -

13.
i
ii
14.
i

+ -

+ -

(+ ) = indicates presence, ( - ) = indicates absence.


A= water, B= chloroform, C= toluene, D= carbon tetrachloride, E= ethyl acetate, F= hexane,
G= ethyl alcohol, H= methanol, I= acetone, J= 2-propanol, K= petroleum ether 60-80 C, L=
2-butanone, M= dichloromethane, N= ethyl ether.40

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 18


4. Shodhana of SA (Purification)

Bhallataka (SA) is reported under upavisha dravya (semi poisonous drugs), in classical
Ayurvedic pharmacopoeias. It is advocated that shodhana (Purificatory procedures) of the
fruits should be carried out before its internal administration. Though there are different
shodhana methods mentioned in Ayurveda, Ayurvedic Pharmacopoeia of India (API)
recommends only one method for the shodhana of Bhallataka fruits. In study, cows urine,
cows milk and brick powder, were used as media. Ayurveda advocates bhallataka after
shodhana (purificatory procedures).Though there are different shodhana methods mentioned
in Ayurveda41

4.1 Effects of Non-Purified (SA) and effect of liquid media in shodhana

In Ayurvedic literature, the synonym Sopha hetu, Spota hetu, agnika are given to this drug
based on its blister causing nature. The oil in the fruit is responsible for the irritation. The
bhallataka fruit contains 90% Anacardic acid and 10% of Cardol. Other chemical constituents
are bhilawanol , semecarpol and anacardol. Recent studies reported that bhilawanols are known
as urushiols. Anacardic acids are closely related to urushiol. Another study reported that the
corrosive juice from the pericarp of the fruit is found to contain catechol, fixed oil and
anacardol (C18H13O3.COOH) to which the corrosive properties of the juice are due to two
phenolic acids C16H15O3.COOH and C14H13O3.COOH. The media gomutra (cows urine) is
reported for its antimicrobial, antibacterial etc. Cows milk is recommended as one of the
antidote for bhallataka blisters. Brick powder is having adsorbent property; by which it absorbs
irritant oil in the fruit.42

4.3 Method of Sodhana

Bhallataka fruits, sunken in water, were randomly taken. The thalamus portion of the fruits
was removed with the help of a steel cutter. Then it was taken in a vessel containing gomutra
(cows urine) and kept for seven days. Every day the fruits were taken out of the media and
washed with water and fresh gomutra was used. On eighth day Bhallataka was washed and
shifted to the vessel containing godugdha (cows milk) and kept for seven days. Each day it
was washed with water and fresh Godugha was added. On 15th day the samples were taken
out of the media and washed with water then shifted to a bag containing brick powder and

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 19


rubbed thoroughly. It was allowed for three days in the bag containing brick powder. On 18th
day it was washed thoroughly with hot water to remove the brick powder in the sample. Later,
the samples were dried properly to remove the moisture and stored in air tight glass container
for further studies. The same shodhana procedure was repeated thrice to standardize the
procedure pharmaceutically.

4.4 Impact on Anacardol Content.

Due to the decorboxylation of the oil, the anacardic acid gets converted into less toxic
anacardol. Decorboxylation process may start right from cutting the fruit itself and will be
catalysed by giving heat/fire Treatment.43 The increased level of anacardol in the shodhita
bhallataka may be due to the decorboxylation of the anacardic acid in the fruits. More
percentage of oil might have got reduced by soaking the fruits in the gomutra and godugda.
The brick powder is having the adsorbing nature, so some percentage of oil may be absorbed
by the brick powder. There are probable chances that some chemical changes might have taken
place due to the various Medias like gomutra, godugda etc used for its purification. Further
studies should be carried out to find out the chemical interactions between the media and the
bhallataka fruits during shodhana procedure. Anacardol in raw bhallataka was 47.51% and
50.62% in processed. Shodhana (purificatory procedure) increases the anacardol level in
shodhita bhallataka fruit samples. More percentage of the anacardol was due to the conversion
of toxic urushiol into Anacardol.44

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 20


5. Methods for Testing (SA) Immunological Factors

The whole animal model is the most classic pharmacological screening model, which is very
important at the aspect of medicine evaluation because it can apparently respond to the efficacy,
side effect and toxicity of medicines in whole. Although this method is high cost and low
efficient, at present it is still a primary way to drug discovery and evaluation. Several in vitro,
in vivo methods of pharmacological screening of medicinal plants having immunomodulatory
activity have been listed.45

5.1 In vitro methods:


1. Inhibition of histamine release from mast cells
2. Mitogens induced lymphocyte proliferation
3. Inhibition of T cell proliferation
4. Chemiluminescence in macrophages
5. PFC (plaque forming colony) test in vitro
6. Inhibition of dihydro-orotate dehydrogenase

5.2 In vivo methods:


1. Spontaneous autoimmune diseases in animals
2. Acute systemic anaphylaxis in rats
3. Anti-anaphylactic activity (Schultz-Dale reaction)
4. Passive cutaneous anaphylaxis
5. Arthus type immediate hypersensitivity
6. Delayed type hypersensitivity
7. Reversed passive arthus reaction
8. Adjuvant arthritis in rats
9. Collagen type II induced arthritis in rats
10. Proteoglycan-induced progressive polyarthritis in mice
11. Experimental autoimmune thyroiditis
12. Coxsackievirus B3-induced myocarditis
13. Porcine cardiac myosin-induced autoimmune myocarditis in rats
14. Experimental allergic encephalomyelitis
15. Acute graft versus host disease (GVHD) in rats
16. Influence on SLE-like disorder in MRL/lpr mice

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 21


6. Experimental Protocol to evaluate (SA) immunomodulatory activity.
Determination of the immunomodulators effects of SA extracts in adjuvant induced arthritic
(AIA) rat model. AIA is an erosive autoimmune polyarthritis involving both humoral and cell
mediated immune responses that resemble human rheumatoid arthritis (RA). At cellular level
immunosuppression occurred during the early phase of the disease. There was mild synovial
hyperplasia and infiltration of few mononuclear cells in SA treated animals. The induction of
nitric oxide synthase (NOS) was significantly decreased in treated animals as compared to
controls. These observations suggest that the herbal extracts caused immunosuppression in
AIA rats, indicating that they may provide an alternative approach to the treatment of arthritis.

6.1 Adjuvant arthritis in Animal (Rats).

6.1.1 Animal Model.


Both male and female albino rats (Sprauge Dawley Strain) 10 rats taken 5 male 5
female.

6.1.2 Development of arthritis.


Rats were injected with 300 l of Complete Freunds Adjuvant (CFA) in the right hind footpad
and left overnight for development of inammation.

6.1.3 Evaluation of arthritis.


Measuring the thickness of inamed ossicular tissue using a dial gauge calliper assessed degree
of arthritis. Severity of inammation was classied using a six-point scale based on
enlargement, erythema and edema of the tissue.

6.1.4. Treatment
To determine the optimum dose, animals were initially treated with 200, 100, 50, 25, 12.5, and
200 mg/kg body weight of the SA Extract separately. Intraperitoneal administration the
according to body weight it should be found to be the optimum dose for immunomodulatory
property, hence used for further studies. All the animals received SA for 25 days.

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 22


6.2. Lymphocyte proliferation assay used to determine the Cell mediated immunity.
Lymphocyte proliferation assay can be performed using mitogen Con-A. Both the treated and
untreated animals were sacriced to remove the spleen in RPMI-1640 medium. Spleen was
crushed and cell suspension was washed with plain medium. Cells were lysed with 0.9%
ammonium chloride and re suspended in complete medium with 10% fetal calf serum. Cells
were cultured at a nal concentration of 3 105cells/100 l/well in triplicate in at bottom
microtiter tissue culture plates. The optimal concentration of 0.05 g/ml (in vitro concentration
titration done earlier) of both the extracts was added to the wells separately and in combination
with Con-A (5.0 g/ml). After 3 days of incubation at 37 C under humidied air supplemented
with 5% CO2 , 1 ci Hthymidine was added to each well. Six to eight hours later cells were
harvested and aspirated on to glass-ber lter papers using NUNC, Automatic Cell Harvester
and the radiolabel incorporated into DNA was counted using LKB auto beta counter.

6.3. Histology of joint tissues.


Rats were sacriced to remove the knee joints. Specimens were xed for 24 h in 2%
glutaraldehyde in phosphate buffer saline, bisected, decalcied and returned to 2%
glutaraldehyde and submitted for routine parafn embedding. Tissue sections were stained
using hematoxylin and eosin stain. The histological ndings were graded on the basis of
synovial hyperplasia and mononuclear cell inltration.

6.4. Measurement of induction of NO in activated macrophages ( Estimation of Nitric


oxide production )
Macrophage activation assay was performed by injecting rats with thioglycolate
intraperitoneally 72 h prior to sacrice. On the day of sacrice, cold RPMI-1640 incomplete
medium was administered intraperitoneally to ush out the activated macrophages. The process
was repeated 810 times to obtain a good yield of macrophages. Once the activated
macrophages were recovered, cells were centrifuged and washed thrice with plain medium,
counted and plated at a nal concentration of 5 105cells/well. Plates were incubated for 4h
at37C in CO2 incubator. Floating cells were removed and the wells were washed twice with
warm medium to avoid the leaching of the adhered macrophages. Cells were then treated with
0.05 g/ml of SA. Culture soup was collected for estimation of NO production by Griess
Reagent method. Briey, 100 l of cell supernatant mixed with 1% sulfanilamide/0.1%
naphthylethylenediamine/2.5% H3PO was incubated at room temperature for 10 min to form
a chromophore. The absorbance was read at 550 nm and NO was measured using NaNO2as

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 23


standard. Quantitation was done against standard curve generated using known quantities of
sodium nitrite.46

6.5. Cell culture


Mononuclear cells were isolated from heparinized venous blood and synovial uid (SF) by
density sedimentation method). Briey, blood/SF was diluted in phosphate buffered saline
(PBS), pH 7.2 and layered carefully on lymphoprep at a ratio of 3:1 and centrifuged at 1800
rpm for 30 min. Cells in the interface layer were carefully separated, washed with PBS thrice
and resuspended in RPMI 1640 supplemented with 25 mM HEPES, 2 mM L-glutamine,
penicillin (100 U/ml), streptomycin (100 _g/ml) and 10%heat inactivated Fetal Bovine Serum
(FBS). Cell concentration was adjusted as per the requirement of the experiment. Cell viability
after 18 h was assessed by mitochondria-dependent reduction of a yellow tetrazolium dye 3[4,5-dimethylthiazol-2yl]-2, 5-diphenyltetrazolium bromide to insoluble purple formazan by
dehydrogenases at the end of 18 h culture. It will showed the Percentage cell viability.

6.6. Cytokine ELISA


Cells were treated with LPS (10 ng/ml) in presence and absence of different doses (1.0, 0.5 and
0.2 mg/ml) of Semecarpus anacardium crude ethanolic extract. The culture was incubated for
18 h at 37 C, 5% CO and supernatants and cells were harvested for ELISA and RNA extraction,
respectively. Pro inammatory cytokines TNF-alpa, IL-1beta, IL-6 and IL12p40 in the culture
supernatants were estimated by commercial sandwich ELISA according to the manufacturers
instruction. Briey, plates were coated with monoclonal capture antibody by overnight
incubation at 42C and blocked with 10% FBS in phosphate buffered saline (PBS) for 1 h.
Samples and recombinant standards were added to the plates and incubated for 2 h. Cytokines
were detected by addition of horseradish peroxidase conjugated streptavidin labelled
antibodies. Color was developed using tetramethylbenzidine (TMB) for 15 min and absorbance
was recorded at 450 nm.47

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 24


7. Reported immunomodulatory activity of Semecarpus anacardium.
Table 4. Reported Immunomodulatory activities of Semecarpus anacardium

S.No Extract : Description


E/
Formulati
on : F
Nut milk Nut Milk Extract in
1
extract : E Aflatoxin B1 -induced
Hepatocellular
Carcinoma
in
Rats.Immunomodulat
ory
activity
was
assessed by measuring
serum
immunoglobulin (Ig)
levels in control and
experimental animals.
Reduced IgG and
elevated IgA and IgM
in the
hepatocellular
carcinoma condition
were returned to near
normal levels in rats
treated
Nut
Immunomodulatory
2
ethanolic
activity of the nut
extract
extracts at 1g/mL in
(endotoxin mononuclear cells of
free):E
production normal.

In vivo models Reported


Dose
or
mechanism of
In vitro models action/Level

Ref
No.

Adult
albino
rats

48

Male Returned
to 200
wistar near
normal mg/kg
levels
of
Reduced IgG
and elevated
IgA and IgM

Humans
cells(Vitro)
-Cell culture
-Cytokine ELISA
-Nitric
oxide
(NO) estimation
-RT-PCR
-Electrophoretic
Mobility
Shift
Assay (EMSA)
Nut milk The components of Adult
Male
extract : E immune system levels albino
wistar
of reactive oxygen rats
species (ROS), namely
Hydroxy
radical,
Superoxide
radical,
and H2O2 were also
measured in spleen,
thymus,
and lymphocytes
Kalpaamr Kalpaamruthaa (KA) Female albino
uthaa :F
is a modified Siddha Sprague

Inhibition of 1g/m
proinflammato L
ry
cytokine
production

49

A significant 150
increase in the mg/kg
level of LPO,
ROS

50

Immunoglobul
ins, and

51

200
mg/kg

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 25


preparation, which has Dawley rats of glycoprotein
been formulated. It Wistar stain
components to
contains Semecarpus
near
normal
anacardium
Linn.
levels
(SA),
Emblica
officinalis (EO), and
honey.
Synergetic
Immunomodulatory
activity
Immunomodulatory
effect
of
Kalpaamruthaa
on
7,12-dimethyl
benz(a)anthraceneinduced mammary
carcinoma studied in
rats
7.1. Balchandran Premalatha et al (1998) has reported Immunomodulatory Activity of
Semecarpus anacardium Linn. Nut Milk Extract in Aflatoxin B -induced Hepatocellular.

Administration of 200 mg kg- S. anacardium nut milk extract brought immunoglobulin levels
back to near normal levels in HCC-induced rats.This indi cates the immunomodulatory effect
of the nut milk extract in a growing tumour, and it wouldbe effective in inhibiting the growth
of an established tumour.No significant change in immuno-globulin levels in drug control
animals suggests that the drug did not produce any immunotoxic effects. The combined
antitumour potency and immuno-modulatory activity of S. anacardium nut milk extract make
it potentially useful in the treatment orprevention of immune-based diseases such as cancers.
7.2. Singh, Divya, et al.(2006) Reported Immunomodulatory activity of Semecarpus
anacardium extract in mononuclear cells of normal individuals and rheumatoid arthritis
patients

The suppressive activity of SA extract demonstrable in normal mononuclear cells was also seen
where peripheral blood and synovial mononuclear cells of RA patients were used. Rheumatoid
arthritis is marked by chronic synovitis and abundance of proinammatory cytokines, TNFalpha IL-1beta, IL-6 and IL-12p40, produced primarily by stimulated monocytes, macrophages
and synovial lining cells

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 26


The suppression of IL-1_ and IL-12p40 along with NF-_B and AP-1, therefore, suggests that
SA may have important constituents that will have benets for this disease.We have also shown
the SA suppressed LPS activated nitric oxide production by the extract in mouse macrophagelike cell,RAW264.7. This is not surprising since nitric oxide production is regulated by iNOS
gene that has promoter regions for NF-KB.NO also plays a role in the pathogenesis of RA and
therefore, its suppression by SA extract adds an additional advantage for its anti-disease
activity

7.3. Ramprasath, et al.(2006) Reported "Evaluation of antioxidant effect of Semecarpus


anacardium Linn. nut extract on the components of immune system in adjuvant arthritis.

Semecarpus anacardium Linn. Nut extract was investigated by studying the extent of lipid
peroxidation and the activities of SAD, CAT, GPx, GSH, and ROS in the lymphocytes and
lymphoid organs (spleen, thymus) in control and experimental rats. Anti-arthriticeffect was
studied from the changes in the paw thickness as a measure of paw edema and arthritic score
in arthritic and drug treated rats. In arthritic rats the extent of LPO and ROS were elevated
profoundly and the antioxidants (SOD, CAT, GPx, GSH) were found to be significantly
decreased. CRP levels and ESR were also found to be significantly increased in arthritic rats.
These changes were brought back to near normal levels on treatment with the drug. No
significant changes were observed in drug alone treated control rats. The paw thickness and
arthritic scores were very much increased in arthritic rats, which was significantly reduced on
treatment with the drug. These effects can be attributed to the presence of flavonoids and other
synergistic components in the drug.
7.4. Dharmendra et al.(2014) Immunomodulatory effect of Kalpaamruthaa on 7, 12dimethyl benz (a) anthracene-induced mammary carcinoma studied in rats."

Kalpaamruthaa (KA) is a modified Siddha preparation, which has been formulated in our
laboratory. It contains Semecarpus anacardium Linn. (SA), Emblica officinalis (EO), and
honey in a definite ratio. The component study of this herbal preparation revealed the presence
of flavonoids, ascorbic acid, polyphenols, tannins, sugars, sterols, etc. Dose-dependent study
of Kalpaamruthaa was carried out in mammary carcinoma-bearing rats, which helped us fix
300 mg/kg body weight as the effective

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 27


This reduction in the levels of glycoprotein components indicates that the drug KA has
the ability to suppress malignancy by modulating the cell transformation, decreasing the degree
of metastasis, inhibiting the progression of growth, and controlling the cancer cell proliferation
and differentiation by causing effective favorable changes in the structure of cell membranes.
This could be due to the cytostabilizing property of the drug and also due to inhibitory action
of flavonoids against carcinogenesis

In this study suggest that DMBA-induced mammary carcinoma is associated with


immune suppression and the drug KA by means of its immunomodulating effect can serve as
a better anticancer agent. The results also suggest that KA is found to be more effective than
SA, which may be due to the amalgamated and additive effects of SA, E.officinalis, and honey
present in the drug.

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 28


8. Toxicological studies of Semecarpus anacardium.
8.1.1.Patwarddhan et al ., 1988 have reported the toxicity of Semecarpus anacardium.
extract
Toxicity by oral route administration of SA with peanut oil was compared against the same
extract emulsified with Tween-80 saline. The traditional way of administration with peanut oil
was found to be safe and upto 25 mg/kg/day x9 days, increase in weight, RBCs & haemoglobin
% was observed without mortality. Same dose with Tween-80 saline was found to have adverse
effects regarding all the parameters with 16.5% mortality. This study support Ayurvedic
method of administration for efficacy without toxicity.
The toxicity of bibba can be controlled by administration with peanut oil or similar
vehicles. Upto 25 mg/kg/day dose bibba can be given safely for therapeutic uses. It can act as
a good hematinic agent and as a general tonic. Ayurvedic method of administration has shown
reduction in toxicity with maintained efficacy.
8.1.2. Vijayalakshmi et al.,2000 have been reported Toxic studies on biochemical
parameters carried out in rats with Serankottai nei, a siddha drug-milk extract of
Semecarpus anacardium nut.
A toxicological study was carried out in rats with a Siddha preparation, milk extracts
of SA. The effect of acute (72 h) and subacute (30 days) treatment of the drug with different
dosage on liver and kidney functions and haematological parameters were studied. The acute
toxicity studies with this drug did not produce mortality at any dose level given (75-2000 mg/kg
body weight). No marked adverse alterations were observed in haematological and biochemical
parameters during the subacute toxicity studies (50, 100, 250 and 500 mg/kg body weight). In
the subacute treatment, the highest dose (500 mg/kg body weight) alone showed a moderate
increase in the level of blood glucose, plasma urea, uric acid, and creatinine. In addition,
alteration in lipid profiles were observed which may be attributed to the ghee preparation of
the drug. Decrease in urinary urea, uric acid and creatinine levels were also observed. Histopathological examination of vital organs showed normal architecture suggesting no
morphological disturbances.The present study shows that the Siddha preparations of S.
anacardium nuts do not induce any toxic manipulation on the biochemical parameters
investigated. From these one can infer and hypothesize that this drug is nontoxic and can be
used as therapeutic agent in treating the reported diseases effectively.

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 29


9. Conclusion
SA reported to be a potent immunomodulator. Scientific studies have proven the efficacious role

in preventive medicine and in the management of chronic degenerative diseases. It would


provide an alternative to conventional chemotherapy for a variety of immunological disorder.
Majorly the evaluation of SA as immunomodulators done by using various models and
techniques such as; Adjuvant arthritis in rats (AIA), Lymphocyte proliferation assay,
Measurement of induction of NO in activated macrophages, Cell culture, Cytokine ELISA.
Recent studies found out that the SANE can inhibit proinammatory cytokine production. The
nut milk extract is effective in restoring the fragility of lysosomal membranes in aflatoxininduced hepatocellular carcinoma. The crude extract also suppressed LPS induced nuclear
translocation. The extract also suppressed LPS activated nitric oxide production in mouse
macrophage cell line. And the maximum tolerated dose 250 mg/kg body weight in 50%
alcoholic extract of the fruit when given interaperitoneally to mice was found to be safe.
Ayurvedic method of administration has shown reduction in toxicity with maintained efficacy.
It also shows a drug interaction with propranolol against isoproterenol induced myocardial
damage in rats.

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 30


10. Future strategies.
Besides mention in classic Ayurveda texts and claims of traditional physicians, it is noted that
evidence through scientific studies is emerging to demonstrate the benefits of SA
immunomodulation. However, the definite lacuna in all research publications reviewed in this
report is the inadequate probing into Ayurvedic science. This has led to superficial correlations,
where the plant drugs have remained as mere candidates for testing against selected
pharmacological

reactions.

Deciphering

deeper

into

Ayurvedic

concept

behind

Immunomodulation can help identify better candidates and models for study.

And it will help for selection of clinical and experimental studies. The later are not adequate at
present, but it is worthwhile to project such evidences to provide lead for further studies. Drug
discovery strategies based on natural products and traditional medicines are remerging as
attractive options. A reverse pharmacology approach, inspired by traditional medicine and
Ayurveda, can offer a smart strategy for new drug candidates to facilitate discovery process
and also for the development of rational synergistic botanical formulations.

With respect to the possible future clinical potential of immunomodulation activity is the
observation that therapeutic dosing with SA inhibited established arthritis in rats. The plantderived immunomodulators thus have tremendous future potential for developing new
pharmaceutical products. Phytochemical analysis of these SANEs is in progress to identify
bioactive molecules responsible for immunomodulatory properties in these plants.

Exploring therapeutic Potential of Bhallataka for Immunomodulatory activity | 31


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