Since the beginning of human civilization man has been dependent on

plant kingdom for medicine besides his basic needs, i.e. food, clothing and

shelter etc. Medicinal plants have been a major source of cure of human

diseases since time immemorial. Ancient man derived more than 90

percent of therapeutic agents from higher plants and much of the

medicinal use of plants seems to have been developed through observations

of wild animals and by trial and error. In 20th century much of the

pharmacopoeia of scientific medicine was derived from the herbal

knowledge of native peoples. Even today, traditional system of medicine is

practiced in many countries and major portion of their therapeutic needs

are obtained from plant drugs. Today, one fourth of the worlds population

depends on traditional medicines. Currently, herbal medicines are in much

demand and their popularity is increasing day by day. About 500 plants

with medicinal use are mentioned in ancient literature and around 800

plants have been used in indigenous systems of medicine. India is a vast

repository of medicinal plants that are used in traditional medical

treatments (Chopra et al., 1956).

Many drugs commonly used today are of herbal origin. Indeed, about 25%

of the prescription drugs dispensed in the United States contain at least

one active ingredient derived from plant material. Many drugs listed as

conventional medications were originally derived from plants such as,

salicylic acid (a precursor of aspirin), was originally derived from white

willow bark and the meadowsweet plant. Quinine which is obtained from

Bark of Cinchona is the source of antimalarial drug similarly; Vincristine

Evaluation of some plant constituents for immunomodulatory effect 34
which is used to treat certain types of cancer
is obtained from
Vinca rosea
. The opium poppy (
Papaver somniferum
) yields morphine and codeine which have antidiarrhial and antitussive
properties. Even today, morphine-the most important alkaloid of the opium
poppy-remains the standard against which new synthetic pain relieves is
measured.
3.1. HISTORY OF HERBAL MEDICINE

Herbal medicines have been used by man since ancient time. The Ayurveda

is the known oldest system of medicine from India, which describes

medicinal plants with their therapeutic potential against diseases.

Description of medicinal plants can be traced in China (4000 B.C.). In the

ancient time, Greek and Egypt used plants in the form of medicines.

Aristotle (380 B.C.) and Theophrastus (380 B.C.) described medicinal

plants in their book, Historia plantarum. The Greek physician Galen (AD

129200) devised the first pharmacopoeia describing the appearance,

properties and use of many plants of his time (Patwardhan et al., 2004).

Hippocrates (in the late fifth century B.C.) mentioned 300 to 400 medicinal

plants. In the first century A.D., Dioscorides wrote De Materia Medica, a

medicinal plant catalog which became the prototype for modern

pharmacopoeias. The Bible offers descriptions of approximately 30 healing

plants (Cowan, 1999).

Plant based drugs provide outstanding contribution to modern

therapeutics such as, Vinblastine isolated from the Catharanthus roseus is

used for the treatment of neck cancer (Farnsworth et al., 1967). Vincristine
Evaluation of some plant constituents for immunomodulatory effect 35
is recommended for acute lymphocytic leukemia in
childhood, advanced stages of hodgkins, lymphosarcoma, small cell lung,
cervical and breast cancer (Farnsworth and Bingel, 1977). Stafford, (1991)
informed that recent phytochemical analysis of plants, used in folklore for
the treatment of cancer, which has yielded a number of compounds with
antitumor activity. Plants from many families have also recently been
shown to accumulate alkaloids with anti-HIV activity. In 1860, a German
chemist Carl Koler isolated cocaine, the chemical responsible for the
biological activity. He found that cocaine could act as a local anesthetic in
eye surgery. As the years passed, scientists observed that cocaine
paralyzed nerve endings responsible for transmitting pain. As a local
anesthetic, it revolutionized several surgical and dental procedures. In
1891, an enzyme that broke down proteins (bromelain) isolated from the
fresh juice of pineapple was found to break down blood clots (Patwardhan
et al
., 2004).
Penos, (1983) estimated that 20,000 plant species are used for medicinal

purposes. Farnsworth et al., (1985) reported that more than 80% of people

in Asia are still dependent on traditional and folk remedies for day-to-day

life. Duke, (1990) estimated that the value of those drugs containing

compounds from plant species is about $ 10 billion in the United States

alone. 74% of 119 plant derived drugs were discovered as a result of

chemical studies to isolate the active substances responsible for their

traditional use (Farnsworth et al., 1985). At least 25% of medical

prescriptions in recent times contain one active compound from plant

species.
Evaluation of some plant constituents for immunomodulatory effect 36
There are over 750,000 plants on earth, but only a very few of the
healing herbs have been studied scientifically. Modern pharmacology looks
for one active ingredient and seeks to isolate it. In recent time most of the
research on plants continues to focus on identifying and isolating active
ingredients, rather than studying the medicinal properties of the whole
plants. Many plants have been evaluated for various therapeutic properties
such as anthelmintic, antioxidant, hepatoprotective, immunomodulatory,
anti-inflammatory, antitumor, antimicrobial etc (Chandrashekhar
et al
., 2008; Guerra
et al
., 2003; Chatterjee and Pal, 1984; Gupta
et al
., 2004; Singh and Gupta, 2008).
3.2. MEDICINAL PLANTS WITH ANTHELMINTIC ACTIVITY

Parasitoses have been of concern to the medicinal field for centuries and

the helminthes still causes considerable problems for human beings and

animals. Helminthes are recognized as a major problem to livestock

production (Adejimi and Harrison, 1997). Parasitic helminthes affect

human being and animals by causing considerable hardship and stunted

growth. Helminthic infections are now being recognized as cause of many

acute as well as chronic ill healths among the various human beings as

well as cattles. More than half of the population of the world suffers from

infection of one or the other and majority of cattles suffers from worm

infections (Dwivedi, 2004). Anthelmintics are drug that either kill

(vermicides) or expel (vermifuge) infesting helminthes (Tripathi, 2004).

Synthetic drugs are used to control such kind of infections but due to their

high cost and side effects, the development of more effective and less
Evaluation of some plant constituents for immunomodulatory effect 37
expensive natural drug production are main concern based on
medicinal properties of the plants. Traditional system of medicine reports
the efficacy of several natural plants in eliminating worms (Basuri and
Modi, 2011). The plants are known to provide a rich source of botanical
anthelmintics (Satyavati
et al
., 1976; Lewis and Elvin-Lewis, 1977). A number of medicinal plants have
been used to treat parasitic infections in man and animals (Nadkarni,
1954; Chopra
et al
., 1956; Akhtar
et al
., 2000) (Table 3.1).
Evaluation of some plant constituents for immunomodulatory effect 38
Evaluation of some plant constituents for immunomodulatory effect
39
 Table 3.1: Plants with anthelmintic activity
Plant Part Ext Organ Refere
Name use ract ism nces
d used
Satyan
arayan
Butea
Ascar rao
mono See
dia and
sperm ds
galli Krishn
a
ainah,
1982
Fres
Calotr Pheret Shivka
h,
opis Late ima r and
Aqu
procer x posth Kumar
eou
a uma , 2003
s
Capp Roo Pheret
Eth Mali et
aris t ima
anol al.,
decid bar posth
ic 2004
ua k uma
Evolv Who Pheret
Eth Dash
ulus le ima
anol et al.,
alsino plan posth
ic 2002
ides t uma
Fascio
la
gigant
ica,
Gyan
Lea Taeni Ajaiyeo
andro Met
ves a ba et
psis han
Ste solium al.,
gyana olic
m , 2001
ndra
Pheret
ima
posth
uma
Taeni Szewez
Melia Eth
Dru a uk et
azeda anol
pe solium al.,
rach ic
, 2003
Pheret
Cru ima
Ste
Mimu de posth Mali et
m
sops alco uma, al.,
bar
elengi holi Ascar 2007
k
c dia
galli
Mogh Roo Cru Yadav
Ascari
ania t de et al.,
s
vestit tube extr 1992
suum
a r act
Pheret
ima
Aqu posth
Neola
Ste eou uma, Gunas
marck
m s, Ascar ekaran
The essential oil of
Piper betle
has revealed anthelmintic activity against the earthworm
Pheretima posthuma, in vitro
(Ali and Mehta
,
1970)
.
Vigar, (1984) tested the essential oil obtained from
P. betle
`Sagar Bangla' against tapeworms and reported that the essential oil was
found to be better than the standard piperazine phosphate, where the
activity against hookworm was found to be greater than the reference drug
hexyl resorcinol. The essential oils of
Anacardium occidentale
and
Callistemon viminalis
have also been found to possess anthelmintic activity against earthworms,
tapeworms and hookworms (Garg and Kasera, 1982). The drupe extracts of
Melia azedarach
(Meliaceae) growing in Argentina was tested against a tapeworm and
earthworm showed better activity against tapeworms than the standard
piperazine phosphate, which is used for the treatment of Cestoda infections
(Szewczuk
et al
., 2003). The active principle, solamargine, isolated from the ripe berries of
Solanum viarum
was also found to be effective against
Microfilariae
and adults
S. cervi
(Sinha, 2005).
The research findings showed that Tetradenia riparia, Cassia occidentalis,

Carica papaya, Momordica foetida and Erythrina abysinnica may be of

value in the treatment of helminthiasis (Wasswa and Deogracious, 2006).

In other case crude extract of Momordica charantia showed the significant

anthelmintic property (Das et al., 2006). Similarly, the seed oils of

Gynandropsis gynandra, Impatiens balsamina, Cedrus paniculata, Embelia

ribes and Mucuna pruriens were investigated for their anthelmintic property

against Pheritima posthuma, where all the oils exhibited moderate to

Evaluation of some plant constituents for immunomodulatory effect 40
significant anthelmintic activity, while Embelia
ribes
showed the best anthelmintic activity (Jalalpure
et al
., 2007). Nirmal
et al
., (2007) reported the anthelmintic properties of extract of
Pongamia glabra
. Ademola
et al
., (2007) reported the anthelmintic property of ethanolic and aqueous
extracts of
Spigelia anthelmia
against different ovine gastrointestinal nematodes. Extracts of the aerial
parts of
Cansjera rheedii
(Opiliaceae) were also reported to show significant activity against
Pheretima posthuma
(Mounnissamy
et al
., 2008). Similarly, the crude aqueous extracts of
Ficus racemosa
(Moraceae) were evaluated for anthelmintic activity using adult earthworms
and which showed wormicidal activity (Chandrashekhar
et al
., 2008). Crude alcoholic extract and its different fractions (pet ether, ethyl
acetate and methanol) of
Clitoria ternatea
also possess anthelmintic activity (Khadatkar
et al
., 2008). Mali and Wadekar, (2008) reported that alcoholic and aqueous
extracts of the roots of
Baliospermum montanum
possess significant anthelmintic activity against
Pheretima posthuma
and
Ascardia galli.
Qureshi et al., (2009) also reported that alcoholic extract of leaves of Smilex

zeylanica possess significant anthelmintic activity against Pheretima

posthuma. Deore et al., (2009) observed significant anthelmintic activity of

alcohol and aqueous extracts from the seeds of Cassia tora against

Pheretima posthuma and Ascardia galli. In other case, crude aqueous and

hydro alcoholic extract of the leaves of Chenopodium ambrosioides,

Lawsonia inermis and seeds of Jatropha curcas were tested for anthelmintic

activity against eggs and adult Haemonchus contortus, where both extracts

of C. ambrosioides and J. curcas were found to inhibit the hatching of eggs

Evaluation of some plant constituents for immunomodulatory effect 41
(Tadesse and Mirutse,
2009). Aqueous and ethanolic extract of
Solanum surattense
showed better anthelmintic activity than the standard drugs while aqueous
extract showed better anthelmintic activity in comparison to the ethanolic
extract of
Solanum surattense
(Nayak
et al
., 2009). Methanolic and aqueous extracts of
Euphorbia thymifolia
also showed significant anthelmintic activity against
Pheretima posthuma
and
Ascardia galli
(Kane
et al
., 2009).
Kosalge and Fursule, (2009) investigated that the aqueous extract of

Thespesia lampas roots possess vermicidal anthelmintic activity against

earthworms (Pheretima posthuma), tapeworms (Raillietina spiralis) and

roundworms (Ascardia galli). Ethanolic extracts of Moringa oleifera and

Vitex negundo were tested against Indian earthworm (Pheritima posthuma),

where both the plants showed significant anthelmintic activity (Rastogi et

al., 2009).

John et al., (2009) reported the significant anthelmintic property of the

methanolic extract and its ethyl acetate fraction of Cassia tora leaves

against Indian adult earthworm (Pheretima posthuma). Hydroalcoholic

extracts of aerial parts of Andrographis paniculata, Cajanus cajan and

Silybum marianum and their combinations were evaluated for anthelmintic

properties against Indian adult earthworms (Pheretima posthuma), where A.

paniculata showed better activity while its combination (1:1) with S.

marianum extract was found to be the most potent (Singh et al., 2009).

Pandey et al., (2010) examined the methanolic extract of Ruta graveolens L.

Evaluation of some plant constituents for immunomodulatory effect 42
leaves for anthelmintic activity against Indian earthworms
(
Pheretima posthuma
) and it showed significant anthelmintic activity.
Singh et al., (2011) evaluated hydroalcoholic extracts of Curcuma longa,

Zingiber officinale and combination of Curcuma longa and Zingiber officinale

rhizome extracts (1:1) for their anthelmintic activity against Pheretima

posthuma (Indian earthworm). It was observed that among the two drug

extracts, Curcuma longa showed maximum vermifuge activity while,

maximum vermicidal activity was shown by Zingiber officinale extract. The

combination of hydroalcoholic rhizome extracts of Curcuma longa and

Zingiber officinale also showed a significant anthelmintic activity.

3.3. MEDICINAL PLANTS WITH ANTIMICROBIAL ACTIVITY

Many efforts have been made to discover new antimicrobial compounds

from various kinds of sources such as micro-organisms, animals, and

plants (Table 3.2). One of such resources is folk medicines. Systematic

screening of them may result in the discovery of novel effective compounds

(Tomoko et al., 2002). The increasing prevalence of multidrug resistant

strains of bacteria and the recent appearance of strains with reduced

susceptibility to antibiotics raises the specter of untreatable bacterial

infections and adds urgency to the search for new infection-fighting

strategies (Sieradski et al., 1999).

Evaluation of some plant constituents for immunomodulatory effect 43

Evaluation of some plant constituents for immunomodulatory effect
44
 Table 3.2: Plants with Antimicrobial Activity
Plant Part Ext Activ Referen
Name use ract ity ces
d
Phongpa
Cassi Met Antif
ichit et
a Leaf han unga
al.,
alata olic l
2004
Phongpa
Cassi Met Antif
ichit et
a Leaf han unga
al.,
fistula olic l
2004
Curcu Singh
Met Anti
ligo and
Root han micr
orchio Gupta,
olic obial
ides 2008
Curcu Antib Singh et
Rhiz
ma - acteri al.,
ome 2002
longa al
Gymn
Eth Antib Satdive
ema
Leaf anol acteri et al.,
sylves
ic al 2003
tre
Leaf
Hibisc
,
us Eth Antib Baqir et
Ste
rosa anol acteri al.,
m,
sinen ic al 1994
Flo
sis
wer
Marru Who Masoodi
Met Antib et al.,
bium le
han acteri 2008
vulgar plan
olic al
e t
Leaf
Neriu ,
Eth Antib Baqir et
m Ste
anol acteri al.,
indicu m,
ic al 1994
m Flo
wer
Orcho Ihsan et
Pet Antib
rus al.,
ethe acteri
olitori 2007
r al
us
Semp Stojicevi
ervivu Antif c et al.,
m Leaf - unga 2008
marm l
oreum
Senna Leaf Eth Antib Doughar
siame anol acteri i and
a ic al okafor,
2008
Ste
Xanthochymol, an active principle of
Garcinia xanthochymus
(Guttiferae) demonstrated antimicrobial activity (Baslas and Kumar, 1980).
A study on antimicrobial compounds was done evaluating extracts from
120 plant species of Brazil from 28 different families. It was documented
that 81 extracts obtained from 58 plants were active against
Staphylococcus aureus
and five extracts from four other plants inhibited the growth of
Pseudomonas aeruginosa
(Santos
et al
., 1990).
In Argentina, 122 known plant species were used for therapeutic

treatments. It was documented that among the compounds extracted from

these plants, 12 inhibited the growth of Staphylococcus aureus, ten

inhibited E. coli, and four inhibited Aspergillus niger (Anesini and Perez,

1993). Clausenol a carbazole alkaloid isolated from alcoholic extract of

stem-bark of Clausena anisata (Rutaceae) also showed antibacterial activity

(Chakarborty et al., 1995). It has been reported that aqueous and ethanolic

extracts from plants used in allopathic medicine are potential sources of

antiviral, antitumor and antimicrobial agents (Chung et al., 1995; Vlietinck

et al., 1995). El-Shazly et al., (2002) found that the essential oil of aerial

parts of Tanacetum santolinoides showed strong in vitro activity against

Escherichia coli, Bacillus subtilis and Candida albicans. Natarajan et al.,

(2003) also reported significant antimicrobial activity of the leaves and seed

extracts of Azadirachta indica against various dermatophytes.

Antimicrobial activity of aqueous and ethanolic extracts of Allium sativum,

Zingiber officinale and citrus aurentifolia was tested against Staphylococcus

Evaluation of some plant constituents for immunomodulatory effect 45
aureus, Escherichia coli and Salmonella spp. and it was
found that their combination showed good activity against
S. aureus
(Onyeagba
et al
., 2004).
Ijeh et al., (2005) reported the strong significant antimicrobial activity of

aqueous extract of Ocimum gratissimum and Xylopia aethiopica against

Pseudomonas aeruginosa, Streptococcus fecalis, Escherichia coli and

Lactobacilli. Mullika et al., (2005) reported the antimicrobial effects of Thai

medicinal plants (Senna alata, Eupatorium odoratum, Garcinia mangostana

and Barleria lupulina) against acne inducing bacteria (Propionibacterium

acne and Staphylococcus epidermis) by using disc diffusion and broth

dilution methods. The data indicated that crude extracts had strong

inhibitory effects on Propionibacterium acne and Staphylococcus epidermis.

The antimicrobial activity of crude and methanol extract of Terminalia

bellerica dry fruit was tested against 9 human microbial pathogens. These

pathogens were found to be highly sensitive to the methanol extract than

crude extract and it was also observed that the dry fruit of Terminalia

bellerica possesses potential broad spectrum antimicrobial activity

(Elizabeth, 2005). Sapindus emarginatus, Hibiscus rosa-sinensis, Mirabilis

jalapa, Rheo discolor, Nyctanthes arbortristis, Colocasia esculenta,

Gracilaria corticata, Dictyota sps., and Pulicaria wightiana were also tested

against 6 bacterial strains (Pseudomonas testosteroni, Staphylococcus

epidermidis, Klebsiella pneumoniae, Bacillus subtilis, Proteus morganii, and

Micrococcus flavus). Sapindus emarginatus showed strong activity against

the tested bacterial strains (Nair et al., 2005). Iroggbu and Nkere, (2005)

evaluated the antibacterial property of ethanolic stem bark extracts of

Evaluation of some plant constituents for immunomodulatory effect 46
Picralima nitida
against pathogenic bacteria by using agar well and macro broth dilution
methods as they showed high bactericidal effects to the crude ethanolic
extract.
Kumar et al., (2006) screened 61 Indian medicinal plants belonging to 33

different families for their antimicrobial properties. All the plant extracts

showed significant antimicrobial activity. Defatted acetone and methanolic

extracts of Bauhinia variegata were also reported to have antimicrobial

activity (Parekh et al., 2006). Byeoung et al., (2006) isolated a compound 2-

(hydroxy methyl) anthraquinone from Tabebuia impetiginosa, which

exhibited strong antimicrobial activity against Helicobacter pylori. The

crude ethanolic extract and other partially extracted fraction of the barks of

Xylocarpus granatum showed significant antimicrobial activity against

Staphylococcus epidermis, Staphylococcus aureus, Shigella boydii, and

Proteus spp. and moderate activity against Escherichia coli, Streptococcus

pyogenes, and no activity against Shigella dysentery, Enterococci,

Salmonella typhi ( Alam et al., 2006).

Mi-Youn et al., (2007) isolated 5-hydroxy-1,4-naphthoquinone from

Caesalpinia sappan heartwood and tested for the growth effects towards

intestinal micro-organisms. Their results revealed that 5hydroxy-1,4-

naphthoquinone had selective growth inhibition against Clostridium

perfringens. Ethanolic extracts of Hemidesmus indicus (roots), Eclipta alba

(fruits), Coscinium fenestratum (stems), Curcubito pepo (seeds), Tephrosia

purpurea (roots), Mentha piperita (leaves), Pongamia pinnata (seeds),

Evaluation of some plant constituents for immunomodulatory effect 47
Symplocos racemosa (barks), Euphorbia hirta
(roots)
, Tinospora cordifolia
(roots)
, Thespesia populnea
(roots)
,
and
Jasminum officinale
(flowers) were tested against
Propionibacterium acnes
and
Staphylococcus epidermidis
(pus-forming bacteria in acne) for antimicrobial activities. Among those
Hemidesmus indicus, Coscinium fenestratum, Tephrosia purpurea,
Euphorbia hirta, Symplocos racemosa, Curcubito pepo
and
Eclipta alba
had shown strong inhibitory effects, however, the
Coscinium fenestratum
extract had a maximum inhibitory effect on
P. acnes
and
S. epidermidis
(Kumar
et al
., 2007). Similarly, the ethyl acetate and n-butanol fractions of ethanolic
extract of
Bacopa monneri
aerial parts were screened for antibacterial and antifungal activities and
both the fractions showed the antimicrobial activity, however, ethyl acetate
fraction was found to be more potent than the n-butanol fraction (Ghosh
et al
., 2007). Ibukum
et al
., (2007) reported the antimicrobial properties of different parts of
Citrus aurentifolia
(lime fruit) against different pathogenic organisms (
Staphylococcus aureus, Salmonella paratyphi, Shigella flexnerii,
Streptococcus faecalis, Citrobacter spp, Serratia spp, Klebsiella pneumonia,
Pseudomonas aeruginosa, Escherichia coli
and fungi
Aspergillus niger
and
Candida albicans
).
Datura metel was investigated in vitro against Ascochyta rabiei (cause of

chickpea blight disease) for antimycotic activity which significantly

suppressed the growth of the target fungal pathogen (Shafique and

Shafique, 2008). Ajayi, (2008) reported that twenty three out of forty eight

Nigerian medicinal plants, when analyzed for antimicrobial property,

Evaluation of some plant constituents for immunomodulatory effect 48
Bacillus spp. and E. coli were found to be most
susceptible ones and the study showed that plants were capable of
producing toxic materials, which may exert some physiological effects on
target organisms. The antimicrobial assay of twenty five Australian native
plants showed inhibiting the growth of one or more bacteria.
B. cereus
was found most susceptible bacteria than
B. subtilis
(Cock, 2008). The antimicrobial activity of aqueous, ethanol and chloroform
extracts of three
Baccharis
species currently used in Northern Chile folk medicine found to be more
active against Gram-positive than Gram negative bacteria, yeast and fungi
tested (Morales
et al
., 2008). Antimicrobial activity of cinnamon essential oil of
Cinnamomum zeylanicum
was studied against
Paenibacillus larvae
. Cinnamaldehyde and eugenol proved to have antibacterial effects against
P. larvae
(Gende
et al
., 2008). Borchatrdt
et al
., (2008) reported that aqueous ethanol extracts of stems, leaves, flowers
and roots of
Betula papyrifera
(Betulaceae),
Centaurea maculosa
(Asteraceae),
Epilobium angustifolium
(Onagraceae),
Hypericum perforatum
(Clusiaceae),
Lythrum salicaria
(Lythraceae), and
Rhus glabra
(Anacardiaceae) inhibited the growth of
Staphylococcus aureus
,
Escherichia coli, Pseudomonas aeruginosa and Candida albicans
. Similarly, Chloroform extract of leaves of
Alternanthera sessilis
also showed antimicrobial activity against some organisms (Jalalpure
et al
., 2008).
Sharma et al., (2009) observed that the ethanol extract of Zingiber officinale

and Punica granatum showed strong antibacterial activity against

Escherichia coli, ethanol extracts of Terminalia chebula and Ocimum

Evaluation of some plant constituents for immunomodulatory effect 49
sanctum exhibited antibacterial activity against Klebsiella
pneumonia
, similarly ethanol extract of
Cinnamomum cassia
showed maximum antibacterial activity against
Pseudomonas aeruginosa
while ethanol extract of
Azadirachta indica
and
Ocimum sanctum
exhibited antibacterial activity against
Enterococcus faecalis
.
3.4. MEDICINAL PLANTS WITH ANTIOXIDANT ACTIVITY

Reactive oxygen species (ROS), sometimes called active oxygen species, are

various forms of activated oxygen, which include free radicals such as

superoxide ions (O2-) and hydroxyl radicals (OHO), as well as non free-

radical species such as hydrogen peroxide (H 2O2) (Halliwell, 1995;

Robinson et al., 1997 ; Squadriato and Pelor, 1998). In living organisms

various ROSs can form in different ways, including normal aerobic

respiration, stimulated polymorphonuclear leukocytes and macrophages,

and peroxisomes. These appear to be the main endogenous sources of most

of the oxidants produced by cells. Exogenous sources of free radicals

include tobacco smoke, ionising radiation, certain pollutants, organic

solvents, and pesticides (Halliwell and Gutteridge, 1989; Davies, 1994).

Free radicals can cause lipid peroxidation in foods, which leads to their

deterioration (Sasaki et al., 1996; Miller et al., 1995). In addition, reactive

oxygen species have been implicated in more than 100 diseases, including

malaria, acquired immunodeficiency syndrome, heart disease, stroke,

arteriosclerosis, diabetes, and cancer (Tanizawa et al., 1992; Hertog et al.,

1993; Alho and Leinonen, 1999; Duh, 1998).

A potent scavenger of these free radical species may serve as a possible

Evaluation of some plant constituents for immunomodulatory effect 50
preventive intervention for free radical mediated diseases (Ames
et al
., 1995). Currently, the possible toxicity of synthetic antioxidants has been
criticized. It is generally assumed that frequent consumption of plant-
derived phytochemicals from vegetables, fruit, tea, and herbs may
contribute to shift the balance toward an adequate antioxidant status
(Halliwell, 1995). Thus, interest in natural antioxidant, especially of plant
origin, has greatly increased in recent years (Jayaprakash and Rao, 2000).
Recent studies showed that a number of plant products including
polyphenolic substances (e.g., flavonoids and tannins) and various plant or
herb extracts exert potent antioxidant actions (Kiselova
et al
., 2006; Sheng
et al
., 2008; Iqbal
et al
., 2009).
Yildirim et al., (2001) evaluated the antioxidant activities of water, ethanol,

and ether extracts of the leaves of Cydonia vulgaris by the thiocyanate

method. It was reported that antioxidant activity of water extract was

increased with the increasing amount of extract (200 g1000g) added to

the linoleic acid emulsion. Ether extract was the most effective antioxidant

among the extracts. Like antioxidant activity, the reducing power of water

extract was concentration dependent. However, ethanol extract was the

highest in reducing power, while ether extract was the lowest. The results

obtained in the present study indicate that the leaves of Cydonia vulgaris

are a potential source of natural antioxidants.

Antioxidant activity of methanol extracts of rhizome of Curculigo orchioides

was evaluated using carbon tetrachlorideintoxicated rat liver (Venukumar

and Latha, 2002). Javanraedi et al., (2003) investigated antioxidant activity

Evaluation of some plant constituents for immunomodulatory effect 51
and total phenolic content of Iranian Ocimum
basilicum
and found that a linear positive relationship existed between the
antioxidant activity and the total phenolic content.
Karou et al., (2005) screened Combretum micranthum, Khaya senegalensis,

Petrocarpus erinaceus and Sida acuta for antioxidant and antibacterial

activities of polyphenols; different polyphenolic contents showed potent

antioxidant activity. Yerra et al., (2005) investigated the chemical

composition and antioxidant activity of different extracts obtained from

Teucrium species and found the presence of flavonoids, luteolin, apigenin

and diosmetin. Teucrium species possess free radical and hydroxyl radical

scavenging as well as antioxidant activity in vitro. Adriana et al., (2005)

reported the antibacterial and antioxidant activities of ethanol extract from

Paullinia cupana seeds, where antioxidant activity and total catechol

content was 0.96. Yerra et al., (2005) evaluated antioxidant activities of

methanol extract of Mucuna pruriens seeds. The total polyphenolic content

in the extract was tested using Folin-Ciocalteu reagent, which was 33.04

mg/g when compared to gallic acid.

Bektas et al., (2006) screened two species of Pimpinella (Pimpinella

anisetum and Pimpinella flabellifolia) for their antioxidative properties using

DPPH free radical scavenging and -carotene/linoleic acid methods. P.

anisetum exerted greater antioxidant activity than Pimpinella flabellifolia.

Sreevidya et al., (2006) evaluated the free radical scavenging potential of

Chlorophytum tuberosum. The extract has been found to possess

antioxidant activity in all the tested models.

Evaluation of some plant constituents for immunomodulatory effect 52
Buppachart et al., (2007) also reported the antioxidant activity of
the ethanolic extract of dried
Schefflera leucantha
leaves. The extract was found to possess the significant antioxidant
activity. Heo
et al
., (2007) evaluated methanol extracts of the aerial parts of 11 Korean
medicinal salad plants for their antioxidant activity. The highest total
phenolic content of the methanol extracts was found in
Aster scaber
, followed by
Ixeris dentate
,
Aster yomena
and
Sedum sarmentosum
. Methanol extracts of
Ixeris dentate
and
Aster scaber
exhibited the highest DPPH radical scavenging activity. Siddhuraju and
Manian, (2007) observed the antioxidant and free radical scavenging
capacity of dietary phenolic extracts from horse gram (
Macrotyloma uniflorum
) seeds. All extracts exhibited good antioxidant activity (53.3-73.1 %)
against the linoleic acid emulsion system but were significantly lower
(P<0.05) than the synthetic antioxidant, butylated hydroxyl anisole (BHA)
(93.3%).
Aneta et al., (2007) investigated the total equivalent antioxidant capacity

(TEAC) and phenolic contents of 32 species extracts from 21 botanical

families grown in Poland. Herbs, Syzygium aromaticum and Epilobium

hirsutum showed highest TEAC values. A positive relationship between

TEAC values and total phenolic content was measured by HPLC.

Khalaf et al., (2008) screened the methanolic crude extracts of some

commonly used medicinal plants for their free radical scavenging

properties using ascorbic acid as standard antioxidant. Free radical

scavenging activity was evaluated using 1,1-diphenyl-2-picrylhydrazyl

(DPPH) free radical. The overall antioxidant activity of green tea (Camellia
Evaluation of some plant constituents for immunomodulatory effect 53
sinensis) was the strongest, followed in descending order by black tea
(
Camellia sinensis
),
Eugenia caryophyllus
,
Piper cubeba
,
Zingiber officinale
and
Piper nigrum
,
Trigonella foenum graecum
and
Elettaria cardamomum
showed weak free radical scavenging activity with the DPPH method. All the
methanolic extracts exhibited antioxidant activity significantly.
Manian et al., (2008) investigated aerial root of Ficus benghalensis and

stem bark of F. racemosa for their antioxidant activity and compared with

Camellia saneness. All the extracts exhibited antioxidant activity against

the linoleum acid and emulsion system. Ozsoy et al., (2008) investigated

the antioxidant activity of water, infusion, ethanol and ethyl acetate

extracts of Smilax excelsa leaf. All extracts showed the presence of good

total phenolic and flavonoid contents inhibiting lipid peroxidation, radical

scavenging and ironchelating activities, which was considered as a

significant natural antioxidant source.

Audipudi and Chakicherla, (2010) investigated methanol and chloroform

extracts of Gmelina arborea for their possible antioxidative activities by

using 2, 2 diphenyl picryl-1-hydrazyl (DPPH) and -carotene bleaching

methods. Methanol and Chloroform extracts were found to be better when

compared to synthetic antioxidant Butylated Hydroxyl Toluene (BHT) in

their antioxidant activity. Methanol extract was found to be better than

chloroform extract in inhibiting the oxidation of linoleic acid.

Evaluation of some plant constituents for immunomodulatory effect 54

3.5.
MEDICINAL PLANTS WITH IMMUNOMODULATORY ACTIVITY
The human immune system has the essential function of protecting the

body against the damaging effects of microbial agents that are pathogenic.

The system comprises innate (non-specific) and acquired (specific)

immunity. Apart from these natural mechanisms, there are additional

factors that stimulate and suppress host immunity. Immunostimulants

enhance the overall immunity of the host, and present a non-specific

immune response against the microbial pathogens and diseases. The use of

plant products as immunostimulants has a traditional history. However,

the isolation of the active principals involved did not gain momentum until

the 19th century (Phillipson, 2001).

Evaluation of some plant constituents for immunomodulatory effect 55

Evaluation of some plant constituents for immunomodulatory effect
56
 Table 3.3: Plants with Immunomodulatory Activity
Plant Par Activity Refere
Name t nces
use
d
Immuno Gokhal
Argyreia Roo
stimulati e et al.,
speciosa ts
on 2003
Calliandr
Immuno Silva
a Lea
stimulati et al.,
pulcherri ves
on 2005
ma
Immuno Nores
Cedrela Lea
modulati et al.,
lilloi ves
on 1997
Wo Immuno Shinde
Cedrus
od supressi et al.,
deodara
oil on 1999
Chloroph Thaku
Immuno
ytum Roo r et al.,
stimulati
borivilian t 2006
on
um
Bafna
Rhi Immuno
Curculigo and
zom stimulati
orchioids Mishra
es on
, 2006
Glycyrrhi Sho Immuno Nose
za glabra ot stimulati et al.,
on 1998
Hai
Glycyrrhi Immuno Nose
ry
za stimulati et al.,
root
uralensis on 1998
s
Hibiscus Immuno Fakeye
Cal
sabdariff modulati et al.,
yx
a on 2008
Da
Orbygny silva
Immuno
a Fru and
stimulati
pharlerat its Parent
on
a e,
2001
Rivera
Panax Roo Adjuvant
et al.,
ginseng t property
2003
Phyllanth Lea Immuno Iganaci
us ves stimulati ao et
tenellus on and al.,
antitumo 2001
r
Piper Fru Immuno Sunila
longum its stimulati and
on Kuttan
, 2004
Ste Manos
Pouteria Immuno
m roi et
Plant Par Activity Refer
Name t ences
use
d
Thabr
Immuno
Phyllanth Lea ew et
stimulati
us debelis ves al.,
on
1991
Platycodo Choi
Immuno
n Roo et al.,
stimulati
grandiflor ts 2001
on
um
Tripterygi Immuno Duan
Roo
um suppress et al.,
ts
Root suspension of Janakia arayalpathra was found to have

immunostimulatory properties in mice. It stimulated an increase in

humoral antibody titres and also of antibody secreting spleen cells in the

plaque forming cells assay following immunization with sheep erythrocytes.

It also increased the number of peritoneal macrophages and produced an

increase in delayed hypersensitivity reaction in mice (Subramoniam et al.,

1996).

Ovalbumin immunized mice treated with Azadirachta indica leaf extract

had higher IgG and IgM levels and anti-ovalbumin antibody titres as

compared to control (humoral response). Azadirachta indica also induced

cell mediated response as seen from the enhancement of macrophage

migration inhibition and footpad thickness (Ray et al., 1996). In other case,

the successive methanol extract of aerial roots of Ficus benghalensis was

found to stimulate cell mediated and antibody mediated immune responses

in rats. It also enhanced the phagocytic function of the human neutrophils,

in vitro (Gabhe et al., 2006). Dahanukar and Thatte, (1997) reported that

Evaluation of some
Asparagus plant constituents for immunomodulatory effect 57
racemosus,
Tinospora cordifolia
and
Withania somnifera
protected animals against infections in normal and immunosuppressed
states induced by hemisplenectomy or surgery. Manjrekar
et al
., (2000) showed that both
Tinospora cordifolia
and
T. sinensis
stems possess immunomodulatory activities and have ability to inhibit
cyclophosphamide-induced anemia, the water extract of
T. sinensis
being superior to the water extract of
T. cordifolia
in increasing the WBC count in mice. Similarly, the findings showed that
both hydroalcoholic extract and polyphenolic fraction of
Allium hirtifolium
decreased acquired immunity response in a dose-dependent manner.
However, only polyphenolic fraction of
A. hirtifolium
showed a dose-dependent effect on intrinsic immunity (Jafarian
et al
., 2003).
Haridradi Ghrita (HG) a panchgavya based polyherbal formulation of

Emblica officinalis (4 g), Terminalia chebula (4 g), Terminalia bellirica (4 g),

Azadirachta indica (4 g), Sida cordifolia (4 g), Glycyrrhiza glabra (4 g) and

cow ghee (76 g) showed a significant increase in neutrophil adhesion and

delayed type hypersensitivity (DTH) response, while the humoral response

to sheep RBCs was unaffected. Thus, HG significantly potentiated the

cellular immunity by facilitating the footpad thickness response to sheep

RBCs in sensitized rats (Fulzele et al., 2003). The dried root extract and

isolated quercetin glycoside of Heracleum nepalense significantly increased

the in vitro phagocytic index and lymphocyte viability in all assays and also

showed a significant increase in antibody titer, carbon clearance and

delayed type hypersensitivity in mice (Dash et al., 2006). It was also

Evaluation of some plant constituents for immunomodulatory effect 58
reported that Hemidesmus indicus
extract stimulated the cell proliferation and promoted the release of IgG by
lymphocytes, and also the ADA activity (Kainthla
et al
., 2006).
Gaur et al., (2009) tested the hydro-alcoholic extracts of Hibiscus rosa

sinensis and Cleome gynandra for immunomodulatory activity. The results

showed that Hibiscus rosa sinensis found to possess significant

immunostimulatory action on immune system while hydroalcoholic extract

of Cleome gynandra exhibited significant immunosuppression effect in dose

dependent manner when compared with control group. Immunomodulatory

effect of Aesculus indica in rats was evaluated by testing the humoral

(antibody titre) and cellular (foot pad swelling) immune responses to the

antigenic challenge by sheep RBCs and by neutrophil adhesion test, where

A. indica showed a significant stimulation of the cell mediated immunity,

while no effects on the humoral immunity (Chakraborthy, 2009).

The aqueous extract of Caesalpinia bonducella seeds was tested for its

effect on cell mediated and humoral components of the immune system in

rats. Administration of C. bonducella seed extract produced an increase in

haemagglutinating antibody (HA) titer and showed significant

immunostimulatory result in delayed type hypersensitivity (DTH) as

compared to control (Shukla et al., 2010).

The immunosuppressive activity of the aqueous extract of fruit of

Lagenaria siceraria consisting of mixture of saponins, flavonoids, tannins,

steroids, phenol and glycosides was studied in mice. The assessment of

immunomodulatory activity was carried out by testing the humoral

Evaluation of some plant constituents for immunomodulatory effect 59
(antibody titre) and cellular (foot pad swelling) immune
responses to the antigenic challenge by sheep RBCs. Oral administration of
extract showed a significant decrease delayed type hypersensitivity
response whereas the humoral response to sheep RBCs was unaffected.
Thus the extract significantly suppressed the cellular immunity by
decreasing the footpad thickness response to sheep RBCs in sensitized
mice. The study demonstrates that the extract shows preferential
suppression of the components of cell-mediated immunity and shows no
effect on the humoral immunity (Sankari
et al
., 2010).
Apart from these works much study has been done on plant extracts and

much is to be done in future. There is no doubt that the potential to

discover new bioactive natural products from plants is great and new drugs

will be created in the near future.

Evaluation of some plant constituents for immunomodulatory effect 60