Boerhaavia diffusa Linn.: A Review on its Phytochemical and Pharmacological Profile
Boerhaavia species have been of keen interest in phytochemical and pharmacological research due to their excellent medicinal values. They are rich sources of alkaloids, steroids and flavones. Boerhaavia diffusa Linn. (Punarnava) has attracted a lot of attention due to its prevalent uses in Ayurvedic system of medicine. It is widely used in jaundice, hepatitis, oedema, oligurea, anemia, inflammation, eye diseases etc. Pharmacologists and clinicians have investigated B. diffusa for all these activities. In some cases the findings support the existing clinical uses. It possesses hepatoprotective, diuretic, anti-inflammatory, anti-stress and immunomodulation, antifertility, antimicrobial, antiviral and insecticidal activities. In conclusion, B. diffusa Linn. contains biologically active compounds that may serve as candidates for new drugs in the treatment and prevention of human and livestock diseases.
Received: April 21, 2011;
Accepted: June 20, 2011;
Published: July 29, 2011
Ayurveda is a native Indian healthcare system which is currently used by million
of people in India, Nepal and Sri Lanka for their day-to-day healthcare needs
(Cooper, 2008; Goyal et al.,
2011). Plants have great potential uses, especially as traditional medicine
and pharmacopoeial drug. A large proportions of the worlds population
depend on traditional medicine because of the scarcity, high costs of orthodox
medicine (Tagboto and Townson, 2001; Hudaib
et al., 2008) and unpleasant side effects (Dalirsani
et al., 2011). A number of natural products have been used as lead
compounds because of specific activity and low toxicity (Sanda
et al., 2011). Medicinal plants have provided the modern medicine
with numerous plant derived therapeutic agents (Pandey et
al., 2010; Evans, 2000; Oladunmoye
et al., 2009). Many plants contain a variety of phytochemicals (Agrawal
et al., 1993; Agrawal and Singh, 2010; Hemlata
et al., 1994; Pandey et al., 1995,
1997; Pandey and Shukla, 2001)
which have found very important applications in the fields of agriculture, human
and veterinary medicine. Natural products play a dominant role in the development
of novel drug leads for the treatment and prevention of diseases (Newman
et al., 2003; Gilani and Atta-ur-Rahman, 2005;
Srivastava et al., 2011).
It is very important to have sufficient knowledge regarding herbs not only because of their widespread uses but also because they have potential to cause toxic reactions or interact with other drugs.
Boerhaavia diffusa Linn. popularly known as Punarnava is
an important rejuvenative drug used in Ayurveda. It is widely distributed throughout
India and flourishes during rainy seasons. The aerial parts then disappear but
revive or sprout again next year (Sivarajan and Balchandran,
1985). Matured whole plant constitutes the drug in Indian Herbal Pharmacopoeia
(Handa et al., 1998).
The aims of present study were to review the chemical constituents of Boerhaavia diffusa Linn. and their biological activities and highlight their potentials as candidates for new drugs that may be of value in the treatment and preventions of human and livestock diseases.
General botanical description and properties of Boerhaavia diffusa:
Boerhaavia diffusa commonly known as Punarnava belongs to
the family Nyctaginaceae. It is a diffusely branched pubescent or glabrous,
prostrate herbs, abundantly occurring as a weed throughout India, up to an altitude
of 2000 m in the Himalayas.
B. diffusa has single, thick deep penetrating tap root bearing few rootlets
occasionally brown (Surange and Pendse, 1972a). Root
is stout and fusiform with a woody root stock. Stems are creeping, many arising
from root stock, swollen at the nodes and often purple.
Leaves are broadly ovate with slightly rounded or pointed apex and rounded
base. The upper surface of the leaf is green but lower surface is white. The
margin is entire and often pink at maturity, flowers are bracteolate umbels
and deep pink in color. They usually contain 2-3 stamens but sometime only one
stamens in each flower is also observed. The fruits are small 2.5 mm long, oval,
oblong, pubescent, simple, achene, bluntly five ribbed with single seed (Handa
et al., 1998).
Microscopically the mature root of B. diffusa shows a complete ring of wood surrounding the ventral vascular region. The secondary concentric zone of wood tissue consists of a few vessels radially arranged and separated by parenchyma which formed the actual wood mass.
The stem shows a single layer of epidermis consisting of cuboidal cells. The epidermis consisting of cuboidal cells. The epidermis is covered from outside by cuticle and large number of glandular hairs. The cortex consists of 1-3 layers of collenchymas cells and several layers of parenchyma. The endodermis is not distinct. Two cells thick layer of pericycle surround the stele. The stele showed several vascular bundles, few bigger ones situated in the center and others forming a ring around the central bundles.
The lamina of the leaf of B. diffusa shows multicellular glandular hairs
on the surface. The epidermis and the hairs of the leaf are covered with a continuous
layer of cuticle. The epidermis at the upper side generally consists of cuboidal
and lower side of the tubular cells. The lamina is transversed by several veins
each surrounded by permanent bundle sheath (Singh and Udupa,
Ethanomedicinal uses of B. diffusa Linn.: B.diffusa has
a long history of uses by indigenous and tribal people and in Ayurvedic or natural
herbal medicines (Dhar et al., 1968). B. diffusa
root extract strengthens, tones and balances the liver (hepatotonic) (Rawat
et al., 1997). It shows hepatoprotective activity (Chandan
et al., 1991; Das and Agarwal, 2011).
The plant is used in epilepsy, pain in abdomen, dysentery, prolapsus ani, fistula
ani and poison of scolopendrids (Jain and Tarafdar, 1970);
in pneumonia (Saxena, 1986); Jaundice (Kumar,
1992; Singh and Ali, 1992; Singh,
1993; Anis and Iqbal, 1994; Sudhakar
and Chetty, 1998); anaemia, (John, 1984; Basak,
1997); as blood purifier (Tripathi et al., 1996);
in enlargement of spleen (Rajwar, 1983); as stomachic,
emetic, laxative, expectorant, diuretic (Jha et al.,1997);
astringent, antiasthmatic, in abdominal pain cough (Das
and Kant, 1988); as anti inflammatory (Kapur, 1993);
tonic, in urinary troubles, ascites, uterine bleeding (Srivastava
et al., 1986) in liver complaints, heart diseases (Rana
et al., 1996) in dropsy, gonorrhoea, oedema, as diuretic (Singh
and Aswal, 1992; Singh, 1993) in haemorrhoids (Singh
et al., 1996) anaemia, colic, thoracic haemorrhage, constipation,
heart disease, oedema, as antidote to rat bite poison and in rabies (Thakur
et al., 1992) in urinary troubles (Maheshwari
and Singh, 1984) as antiasthmatic, in anasarea, ascites (Banerjee
and Banerjee, 1986) oedema, oligurea, as antidote to snake bite (Chandra
et al., 1989). The fruits are used as a diuretic. The seeds are used
as expectorant. carminative, tonic, anthelmintic in lumbago (Tripathi
et al., 1996); jaundice and gonorrhoea (Mishra
and Verma, 1995). The shoot is used in dysuria, oligurea as haematinic and
uterine stimulant (Borthakur, 1996). The leaves are
used in ophthalmic disease (John, 1984; Shah
and Gopal, 1985, 1986); stomach disorder (Biswas
and Ahmed, 1987); cataract (Reddy et al., 1988;
Balajirao et al., 1995); as analgesic, antiasthmatic,
blood purifier, in dropsy, gonorrhoea, jaundica and for hastening delivery (Raj
and Patel, 1978); as laxative and digestant (Yoganarasimhan
et al., 1982; Sudhakar and Chetty, 1998); appetizer
and alexiteric (Chaudhuri et al., 1995); antihypetensive
(Singh et al., 1998); in rheumatism (Rao,
1981); hypotension (Jha and Varma, 1996); indigestion,
abdominal pain, spleenomegaly (Yoganarasimhan et al.,
1979, 1982); as diuretic, in jaundice, gonorrhoea;
in urinary troubles (John, 1984); liver ailments (Gopal
and Shah, 1985); to check post partum haemorrhage (Singh
and Pandey, 1980); for wound healing (Sebastian and
Bhandari, 1984); for itch and eczema (Saxena and Vyas,
Phytochemical constituents isolated from B. diffusa Linn.: Phytochemicals
are natural bioactive compounds found in plants, including the medicinal plants,
fruits, vegetables, flowers, leaves, roots and fibers and they act as a defense
system against diseases or more accurately protect plants against diseases (Krishnaiah
et al., 2009). The therapeutic potentials, including antioxidant,
antimicrobial and anti-carcinogenic properties of higher plants are due to the
presence of secondary metabolites (Canigueral et al.,
2008; Kaur and Arora, 2009). The medicinal values
of these plants lie in bioactive phytochemical constituents that produce definite
physiological actions on the human and animal body. Some of the most important
bioactive phytochemical constituents are the glycosides, alkaloids, flavonoids,
tannins, steroids, terpenoids, essential oils and phenolic compounds (Harbone,
1984; Edeoga et al., 2005; Okwu,
The B. diffusa plant contains a large number of such compounds as flavonoids,
rotanoids, alkaloids, steroids, triterpenoids (Kadota
et al., 1989; Lami et al., 1990, 1991a;
Jain and Khanna, 1989). These biologically active chemical
substance known as secondary metabolites in medicinal plants, form the foundations
of modern prescription drugs (Sofowora, 1993).
In recent decades, there are many reports on the use of medicinal plants. From the studies, it was discovered that the exact amount of active chemical constituent are frequently unknown. In general, one or two markers of pharmacologically active components in herbs and or herbal mixtures are currently employed for: (1) evaluating the quality and authenticity of herbal medicine; (2) identification of single herb or herbal mixtures and (3) assessing the quantitative herbal compositions of a herbal product. It was discovered that multiple constituents are usually responsible for the therapeutic affects of the plants.
These multiple constituents may act synergistically and could hardly be separated
into active parts. Moreover, the herbal constituents may vary depending on the
harvest seasons, plant origins, drying processes and other factors (Walker,
2004). For example the roots of B. diffusa are used for the treatment
of various hepatic disorders. The effect of seasons, thickness of roots and
form of dose (either aqueous or powder) were studied for their hepato-protective
action (Rawat et al., 1997). The results showed
that an aqueous extract (2 mL kg-1) of roots of diameter 1-3 cm,
collected in the month of May (summer) exhibited marked protection of a majority
of serum parameters, viz., SGOT, SGPT, SACP, SALP but not GLDH and bilirubin,
thereby suggesting the proper size and time of collection of B. diffusa
roots for the most desirable results. The studies also showed that administration
of aqueous form of drug (2 mL kg-1) had more hepato-protective activity
than the powder form.
In a preliminary screening, plant revealed presence of sterols (Singh
and Udupa, 1972), β-sitosterol (Srivastava et
al., 1972; Desai et al., 1973) and alkaloids
(Garg et al., 1980). Presence of steroids, sugars
and alkaloids was also reported (Shukla, 1982).
It contains about 0.04% of alkaloid known as punarnavine (C17H22N2O,
mp 236-237°C) (Surange and Pendse, 1972b) and punarnavoside,
an anti fibrinolytic agent. It also contains about 6% of potassium nitrate,
an oily substance and ursolic acid (Kokate et al.,
2005). The green stalk of the plant has also been reported to contain boerhavin
and boerhavic acid.
Hentriacontane, β-sitosterol and ursolic acid along with glucose, fructose
and sucrose were isolated from the root (Misra and Tiwari,
A new C-methyl flavones characterized as 5,7-dihydroxy-6-8-dimethoxy flavones
was reported from root (Gupta and Ahmed, 1984) and designated
as boerhavone (Ahmed and Yu, 1992).
Four new compounds, boerhavisterol, boerhadiffusene, diffusarotenoid and boerhavilanastenyl
benzoate and a known rotenoid, boerhavinone A were isolated from the root and
their structures elucidated as : 9,10-seco-stigmast-5,8 (9)-dien-3β-ol;
9β-lanost-5-en-3-one and 6-methoxy-9,11-dihydroxy-10-methyl-6-a, 12a-dihydrorotenoid,
respectively (Gupta and Ali, 1998). Chemical structures
of all of the active constituents has been presented in Fig. 1.
Many rotanoids have been isolated from the roots of the plant (Kadota
et al., 1989; Lami et al., 1990,
1991b). These include a series of boeravinones viz.,
boeravinone A, boeravinone B, boeravinone C, boeravinone D, boeravinone E and
Punarnavoside, a phenolic glycoside is reportedly present in roots (Seth
et al., 1986). Punarnavoside was later characterized as 2-glucopyrano-4-hydroxy-5-(p-hydroxy
phenyl) propionyldiphenylmethane (Jain and Khanna, 1989).
Bioactive eupalitin 3-O-β-D-galactopyranoside (5,4-dihydroxy 6,7-dimethoxy-flavonal-3-O-β-D-galactopyranoside)
and eupalitin isolated from the alcoholic extract of B. diffusa leaves
(Pandey et al., 2005) exhibited immunosuppressive
Eupalitin 3-O-β-D-galactopyranoside (1>2)-β-D-glucopyranoside
is isolated from ethanolic extract of roots of B. diffusa (Agrawal
et al., 2011).
Four new compounds were isolated from Boerhaavia diffusa namely (i)
(ii) 3,3,5-trihydroxy-7-methoxyflavone (iii) 4,7-dihydroxy-3-methylflavone
and (iv) 3,4-dimethoxyphenyl-1-O-β-D-apiofuranosyl-(1>3)-O-β-D-glucopyranoside
(Maurya et al., 2007).
A new dihydroisofuranoxanthone, methyl 3,10-dihydro-11-hydroxy-1-methoxy-4,6-dimethyl-10-oxo-1H-furo
[ 3,4-b ] xanthenes-3-carboxylate, designated as borhavine, has been isolated
from the benzene extract of the roots of Boerhaavia diffusa L. (Ahmed
and Yu, 1992).
|| Chemical structure of main constituents isolated from Boerhaavia
A metabolite profiling and biological study was undertaken in Boerhaavia
diffusa leaves and roots and substantial differences were found between
the two parts of the plant. The volatile composition was analysed for the first
time using HS-SPME-GC-MS and several compounds including terpenes, phenyl propanoids,
indole compounds, norisoprenoids, among others, were identified. Organic acid
analysis was also performed allowing their characterization in this species
for the first time and oxalic, ketoglutaric, pyruvic, quinic and fumaric acids
were identified. Quantitative differences between two vegetal materials were
found. Additionally, several flavonoids and one phenolic acid were also confirmed
from the roots and leaves of the plant. These are quercetin 3-O-robinobioside,
eupalitin 3-O-galactosyl (1→2) glucoside, Kaempferol 3-O-robinobioside,
eupalitin 3-O-galactoside, quercetin and Caffeoyltartaric acid (Pereira
et al., 2009).
A bioassay guided separation of a methanolic extract obtained from the roots
of B. diffusa L. allowed to isolate five compounds belonging to the class
of rotenoids, out of which three compounds are known boeravinone D, boeravinone
E and a rotenoid 5 along with two novel compounds boeravinone G and boeravinone
H. Boeravinone G, boeravinone E and rotenoid 5 exhibited spasmolytic activity
(Borrelli et al., 2005).
Two known lignans viz., liriodendrin and syringaresinol mono-β-D-glycoside
have been isolated (Lami et al., 1991a).
Two quinolizidine alkaloids identified as punarnavine-I and punarnavine-II
were isolated from root, stem and leaves. The distribution of these alkaloids
was maximum in stem and minimum in root. A relation was established between
the growth process of the plant and biosynthesis of these two alkaloids. The
alkaloidal content was initially low during commencement of pre-reproductive
phase, gradually increased in different plant parts, becoming maximum during
termination phase of reproductive stage (Nandi and Chatterjee,
Pharmacological and biological activities of B. diffusa Linn.: The plant has drawn lot of attention due to following biological activities:
Analgesic and anti-inflammatory activity: The analgesic property of
aqueous extracts obtained from B. diffusa, mainly from the leaf juice
of the plant. The data also confirmed the traditional indications. The mechanism
underlay this analgesic effect remains unknown but the aqueous extract obtained
from leaf juice is endowed with an apparently morphinomimetic central analgesic
property (Hiruma-Lima et al., 2000).
The aqueous and acetone extracts of the root, showed significant anti-inflammatory
activity against carageenan-induced oedema and formaldehyde-induced arthritis
in albino rats. The aqueous extract and an alkaloid significantly inhibited
the increased serum amino transferase activity in arthritic animals similar
to hydrocortisone. Liver ATP phosphohydrolase activity was also increased by
aqueous extract and the alkaloid (Bhalla et al.,
The water insoluble alcoholic extract of different parts of the plant viz.,
root, stem, leaves and flowers and plant was studied for anti-inflammatory activity
against carageenan-induced oedema in rats and for diuretic activity. The root
and leaves were found to be most active (Mudgal, 1974)
and the activity was found maximum during rainy season (Mudgal,
1975). No conclusive difference in diuretic and anti-inflammatory activities
of leaves with or without flowers was observed (Mudgal et
The effect of extract obtained from the root was studied on experimental acute
pyelonephritis in rats. It reduced the inflammatory changes as well as the abscess
formation in kidneys of animals infected with inoculation of Escherichia
coli. It also reduced the bacterial count in the urine samples of infected
animals (Singh et al., 1988).
Hepatoprotective activity: The hydro alcoholic extract of roots of B.
diffusa (HEBD) exhibited a significant protective action of liver evident
by a reduction in elevated levels of serum lysosomal enzymes namely serum Glutamate
Pyruvate Transaminase (SGPT), Serum Glutamate Oxaloacetate Transaminase (SGOT),
Alkaline Phosphate (ALP) in both CCl4 and rifampicin-isonizid induced
hepatotoxicity. Hence HEBD showed a dose dependent hepatoprotective activity
(Desai et al., 2008).
An alcoholic extract of whole plant of B. diffusa given orally exhibited
hepatoprotective activity against experimentally induced carbon tetrachloride
hepatotoxicity in rats and mice. The extract also produced an increase in normal
bile flow in rats suggesting a strong choleretic activity. The extract does
not show any signs of toxicity upto an oral dose of 2 g kg-1 in mice
(Chandan et al., 1991).
The effect of 50% ethanolic extract of roots of B. diffusa on Country
Made Liquor (CML) induced hepatotoxicity was studied in albino rats. B. diffusa
(100 mg/100 g body weight/day) protected the rats from hepatotoxic action of
CML as evidenced by changes in serum alanine Aminotransferase (ALT), Triglycerides
(TG), Cholesterol and total lipid levels in both serum and tissues. Histopathological
studies showed marked reduction in fat deposits in animals receiving B. diffusa
extract along with CML (Rajkumari et al., 1991).
Different extracts and three isolates from extracts were subjected to in
vivo and in vitro testing for antihepatotoxic activity. The petroleum
ether, chloroform and methanol extracts and the total alkaloids (isolated from
the methanolic extract) of the root were reported to slightly lower the increased
SGOT, SGPT, SALP in rats treated with CCI4. The chloroform and the
methanol extracts exhibiting relatively better enzyme lowering effects were
subjected to further fractionation yielding one of the isolates identified as
rotenoid (6, 11-dihydroxy-9-10-dimethyl-4-methoxyrotenoid). The rotenoid was
reported to lower the SGOT and SGPT at a dose of 100 μg mL-1
of the media against CCl4-intoxication on isolated rat hepatocytes.
Similarly, the enzyme lowering action was observed with 500 mg kg-1.
i.p. administration of chloroform and methanolic extracts of the aerial parts.
A steroid androst-5-ene analogue and a flavone, 6, 5'-dimethoxy-5, 7, 3'-trihydroxyflavone,
isolated from the aerial parts, were also subjected to evaluation of antithepatotoxic
activity against CCl4-intoxication by in vitro method. The
steroid significantly lowered SGPT and SGOT at a dose of 200 μg mL-1
of the media, whereas the flavone significantly lowered the SGOT at a dose of
50 μg mL-1 of the media (Chakraborti and
The effects of seasons, thickness of root and form of dose (either aqueous
or powder) of root were studied for their hepatoprotective action in thioacetamide-induced
liver toxicity in rats. The roots of different diameter were collected in three
seasons, rainy, summer and winter. The results showed that an aqueous extract
(2 mL kg-1) of roots of diameter 1-3 cm, collected in the month of
May, exhibited marked protection of majority of serum parameters viz. SGOT,
SGPT, SACP and SALP but not GLDH and bilirubin, thereby suggesting the appropriate
size and time of collection of root for better results. The studies also showed
that administration of aqueous form of drug (2 mL kg-1) had more
hepatoprotective activity than the powder form. This was probably due to the
better absorption of the liquid form through the intestinal tract (Rawat
et al., 1997).
Diuretic: The effect of extracts of red and white varieties of the plant
was studied on diuresis and renal enzymes. Both the varieties exhibited diuretic
activity in toads. Red variety showed comparatively less activity. It inhibited
the activity of kidneys succinic dihydrogenase but showed stimulatory
effect on it in lower doses. Inhibition produced by red variety was less than
that of white variety. It depressed kidney tissue slice respiration but had
no effect on kidney phosphatase. It stimulated the activity of kidney d-amino
acid oxidase. The activity was more in white variety (Chowdhury
and Sen, 1955).
In an experimental study, the aqueous extract of twigs in a dose of 2.5 mL
100 g-1 in rats showed moderate diuretic activity, slightly more
active than potassium acetate (Gujral et al., 1955).
The diuretic activity of water soluble fraction of root and leaves was reported
in male albino rats and dogs. The activity was attributed to the presence of
potassium content in it (Bhide et al., 1958).
The petroleum ether extract of plant exhibited diuretic activity associated
with increased sodium excretion in rats when given in a dose of 250 mg kg-1
orally. The results were compared with polythiazide (Gaitonde
et al., 1974).
The glucosidic compound isolated from plant showed marked diuretic activity
in saline-loaded rats with reference to urea as standard (Haravey,
Punarnava decoction showed good diuretic activity
in rats in a dose of 1 mL 100 g-1 with normal saline on alternate
days for over 15 days period (Singh et al., 1992).
The diuretic action of the plant was shown to be due to β-ecdysone isolated
from the root of B. diffusa (Suri et al.,
Antistress and adaptogenic activity: The extract improved the stress
tolerance by significantly increasing the swim duration and reducing the elevated
WBC, blood glucose and plasma cartisol. Immunomodulatory activity was evaluated
by carbon clearance and delayed hypersensitivity test. The extract significantly
increased carbon clearance, indicating the stimulation of reticuloendothelial
system. The extract also produced an increase in DTH response to SRBC in mice
(Sumantha and Mustafa, 2007).
Immunomodulatory effect: The alkaloidal fraction from the roots of Boerhaavia
diffusa was studied for its effect of cellular and humeral functions in
mice. Orally administration is significantly inhibited SRBC-induced delayed
hypersensitivity reactions in mice. However, the inhibition was observed only
during post-immunisation drug treatment while no effect during pre-immunisation
drug treatment, was observed. The study revealed that the alkaloidal fraction
exhibited in vivo immunostimulatory activity without an in vitro
effect in mice (Mungantiwar et al., 1999).
In a study to evaluate the adaptogenic potential of root, the aqueous extract
of the root powder was studied for its effect on Escherichia coli-induced abdominal
sepsis, macrophage phagocytic activity in mice and on cold and forced swimming
stress in rats. Pretreatment with root powder extract at a dose of 200 mg kg
day-1 orally for 15 days prior to Escherichia coli challenge
produced significant leucocytosis with reduction in mortality in rats and also
significantly increased macrophage phagocytic activity in mice. The plant extract
reversed the stress-induced elevations in the levels of glucose, cholesterol,
SGPT, BUN and reduction in triglycerides (Mungantiwar et
The alkaloidal fraction isolated from the root was investigated for its effect
on plasma cortisol, adrenal cortisol and humoral response in stressed rats.
It exhibited restorative activity against stress-induced changes in plasma and
adrenal cortisol levels. It also significantly augmented the antibody production
in stressed rats as compared to control (Mungantiwar et
Antifertility: The 50% aqueous ethanolic extract of root, when administered
orally in experimental monkeys was found to stop intrauterine contraceptive
device (IUCD)-induced bleeding at 50 mg kg-1 bw (Seth
et al., 1986).
The ethanolic extract of root in a dose of 250 mg kg-1 bw (daily)
p.o. to pregnant albino female rats during the entire period of gestation did
not show any teratogenic effects, as litter size and survival rate of foetuses
were the same a s for the normal control group and no foetal abnormality was
detected (Singh et al., 1991).
Immuno suppressive activity: B. diffusa hexane, chloroform and
ethanol extracts and two pure compounds Bd-I (eupalitin-3-O-h-D-galactopyranoside)
and Bd-II (eupalitin) were evaluated in vitro for their effect on T cell
mitogen (phytohemagglutinin; PHA) stimulated proliferation of human Peripheral
Blood Mononoclear Cell (PBMC), mixed lymphocyte culture, Lipopolysaccharide
(LPS) stimulated nitric oxide production by RAW 264.7, PHA and LPS induced IL-2
and TNF-α production, in human PBMCs, superoxide production in neutrophils,
human Natural Killer (NK) cell cytotoxicity and nuclear translocation of nuclear
factor-k B and AP-1 in PHA stimulated PBMCS. The chloroform and ethanol extracts
inhibited PHA stimulated two way MLR, NK cell cytotoxicity as well as LPS induced
No production by RAW 264.7; the hexane extract showed no activity. Bd-1 purified
from the ethanolic extract at equivalent dose, inhibited PHA-stimulated proliferation
of peripheral blood mononuclear cells, two-way MLR and NK cell cytotoxicity
as well as LPS induced NO production by RAW 264.7 equally or more effectively
than the parent ethanolic extract. Bd-I inhibited production of PHA stimulated
IL-2 at the protein and mRNA transcript levels and LPS stimulated TNF-α
production in human PBMCs; it also blocked the activation of DNA binding of
nuclear factor-k B and AP-1, two major transcription factors centrally involved
in expression of the IL-2 and IL-2R gene which are necessary for T cell activation
and proliferation. Our results report selective immunosuppressive activity of
B. diffusa leaf (Pandey et al., 2005).
A research is also carried out to evaluate the immunomodulatory properties of
this plant extract on various in vitro tests such as human Natural Killer
(NK) cell cytotoxicity, production of Nitric Oxide (NO) in mouse macrophage
cells. RAW 264.7, interleukin-2 (IL-2), tumor necrosis factor-a (TNF-α),
Intracytoplasmic Interferon-g (IFN-Y) and expression of various cell surface
markers on human Peripheral Blood Mononuclear Cells (PBMCs). Ethanolic extracts
of B. diffusa roots inhibited human NK cell cytotoxicity in vitro,
production of NO in mouse macrophage cells, IL-2 and TNF-α in human PBMCs.
Intracytoplasmic IFN-and cell surface markers such, as CD16, CD25 and HLA-DR
did not get affected on treatment with B. diffusa extract. Hence, it
demonstrates immunosuppressive potential of ethanolic extract of B. diffusa
(Mehrotra et al., 2002a).
Antidiabetic activity: A study was carried out to investigate the effects
of daily oral administration of aqueous solution of Boerhaavia diffusa
L. leaf extract (BLEt) (200 mg kg-1) for 4 weeks on blood glucose
concentration and hepatic enzymes in normal and alloxan induced diabetic rats.
A significant decrease in blood glucose and significant increase in plasma insulin
levels were observed in normal and diabetic rats treated with BLEt (Pari
and Satheesh, 2004). Chloroform extract of B. diffusa leaf produced
dose dependent reduction in blood glucose in streptozotocin-induced NIDDM rats
comparable to that of glibenelamide. The results indicate that the reduction
in blood glucose produced by the extract is probably through rejuvenation of
pancreatic beta-cells or through extra pancreatic action (Nalamolu
et al., 2004).
Radioprotective activity: In a study on the effect of the plant in radiation-induced
haemopoietic injury in albino mice, pretreatment (in the dose of 260 g kg-1
bw orally for 21 days) to mice exposed to total body irradiation (6 Gy) for
3 min showed significant increase in Hb and total RBC count. After irradiation,
there was no fall in RBC count and Hb unlike in controls. The study indicated
that the plant had selective effect on the erythroid compartment (Thali
et al., 1998).
Anti-metastatic activity: Administration of Punarnavine (40 mg kg-1
body weight) prophylactically (95.25%), simultaneously (93.9%) and 10 days after
tumor inoculation (80.1%) could inhibit the metastatic colony formation of melano
main lungs. Survival rate of the metastatic tumor-bearing animals were increased
significantly by the administration of Punarnavine in all the modalities compared
to the metastasis bearing untreated control. These results correlated with the
biochemical parameters such as lung collagen hydroxylproline, uronic acid, hexosamine,
serum sialic acid, serum glutamyl transpeptidase and serum Vascular Endothelial
Growth Factor (VEGF) levels and histopathological studies. Punarnavine administration
could suppress or down regulate the expression of MMP-2, MMP-signal-regulated
kinase) and VEGF in the lung tissue of metastasis-induced animals. Punarnavine
could inhibit MMP-2 and MMP-9 protein expression in gelatin zymographic analysis
of B16F-10 cells. These results indicate Punarnavine could inhibit the metastatic
progression of B16F-10 melanoma cells in mice (Manu and
Prophylactic administration of the methanolic extract (0.5 mg dose-1)
inhibited the metastases formation by about 95% as compared to untreated control
animals. There was 87% of inhibition in the lung metastases formation in syngeneic
C57BL/6 mice, when the extract was administered simultaneously with tumour challenge
(Leyon et al., 2005). The total WBC count prior
to irradiation was 7500±500 cells mm-3 which was reduced to
1500±500 cells mm-3 in the irradiated control group on day
9 after radiation exposure. But in th B. diffusa treated group, irradiated
animals showed the lowest count on day 3 after irradiation (4000±400
cells mm-3). Where the count for irradiated control animals was 2100±440
cells mm-3. By day 9, the level reached 6250±470 cells mm-3
in B. diffusa-treated irradiated animals (Manu et
Antioxidant activity: Leaves revealed stronger antioxidant activity
than roots, the first analysis of volatile compounds of a widely used medicinal
plant B. diffusa, using a HS-SPME-GC-MS technique directly into the headspace
of the aqueous extract of the leaves and roots. In addition to phenolics (determined
by HPLC-DAD), the organic acids (HPLC-UV) profile and in vitro antioxidant
and anti acetylcholinesterase activities are described for the first time, providing
further knowledge on this species chemistry and biological potential (Pereira
et al., 2009). Ethanol and methanol extracts were prepared and screened
for in-vitro antioxidant activities using Ferric reducing power and Hydrogen
peroxide scavenging activity. The activity was compared to standard antioxidant
like ascorbic acid. Both the extract showed strong antioxidant activity in both
the methods. Between these two extracts, ethanolic extract has shown better
antioxidant activity as compared to methanolic extract in both the activities
(Rachh et al., 2009).
Antimicrobial activity: The methanol extract of Boerhaavia diffusa
leaves had significant in vitro antimicrobial activity. Hence, further
results revealed that among several pathogenic bacteria, only Staphylococcus
aureus was susceptible for Boerhaavia diffusa. In Boerhaavia diffusa,
maximum inhibition was observed in Staphylococcus aureus followed by
Bacillus megaterium and Bacilus cereus, respectively at 50 μL
concentration (Girish and Satish, 2008).
The alcoholic extract of root showed antimicrobial activity against Staphylococcus
aureus whereas aqueous extract was active against Escherichia coli
and inactive against Staphylococcus aureus (George
et al., 1947). The phosphate buffer and ether extracts of shoot showed
antibiotic activity against Staphylococcus aureas and was inactive against
Escherichia coli (Joshi and Magar, 1952). The
alcobolic extract of the plant showed antibacterial activity against Escherichia
coli in vitro studies (Singh et al., 1974).
The seed exhibited antibacterial activity against Bacillus subtilis,
Pseudomonas cichorii and Salmonella typhimurium but was inactive
against Escherichia coli (Sushil et al., 1997).
The aqueous extract of leaves of B. erecta and B. diffusa were
screened for antibacterial activity against Alkaligenes viscolactis,
Aeromonas hydrophylla. Cytophaga sp., Escherichia coli,
Klebsiella aerogenes, Pseudomonas aeruginosa, Vibrio parahaemolyticus,
Vib damsela, Bacillus cereus and Streptococcus pyogenes.
B. diffusa failed to exhibit antibacterial activity against all the bacteria
but B. erecta was found to be active against Alkaligenes viscolactis,
Bacillus cereus and Streptococcus pyogenes (Perumal
Samy et al., 1999).
The leaf extract dit not exhibit antifungal activity against Microsporum
nanum (Rai, 1987). The aqueous extract of root exhibited
21-50% inhibition of spore germination against Curvularia lunata,
Cylindrocarpon lichenicola, Fusarium solani and Myrothecium
leucotrichum (Gourinath and Manoharachary, 1991).
The fresh leaf extract inhibited germ tube elongation of Drechslera oryzae
(Ganeshan and Krishnaraju, 1995). The aqueous extract
of leaves/stem/flowers/seed/plant exhibited antifungal activity against 3 keratinophilic
fungi; moderate activity against Microsporum gypseum (39.01%), was less
effective against Chrysosporium tropicum (25.63%) and showed minimum
antimycotic activity against Trichophyton terrestre (1.76%) (Qureshi
et al., 1997).
Anti-viral activity: The Boerhaavia diffusa plant is reported
to posses many pharmacological, clinical and antimicrobial properties. Recently
it is observed potent antiviral efficacy of this plant against phytopathogenic
viruses. The antiviral agent isolated from this plant was found to be a glycoprotein
with a molecular weight of 16-20 kDa. Administered by foliar spraying in the
field, this antiviral agent could protect some economically important crops
against natural infection by plant viruses (Awasthi and
Maximum antiviral activity, in each case, was recorded with the aqueous extract
of dried root powder applied before virus inoculation. The active principle
was purified and isolated (Verma et al., 1979).
The roots of B. diffusa are a rich source of a basic protein which is
used for inducing systemic resistance in many susceptible crops against commonly
occurring viruses (Verma and Awasthi, 1979, 1980;
Verma et al., 1979; Awasthi
et al., 1984, 1985, 1989).
This protein or antiviral agent was active against tobacco mosaic virus in Nicotiana
glutinosa, Datura metel, Chenopodium amaranticolor and Nicotiana
tabacum (Ky58 White Burlley and NP31); sunnhemp rosette virus in Cyamopsis
tetragonoloba, Vigna unguiculata and Crotalaria juncea; and
gomphrena mosaic virus in Chenopodium amaranticolor, Vigna unguiculata
and Gomphrena globosa when applied a few hours (2-24 h) before inoculation
by the respective inocula of viruses (Verma and Awasthi,
1979; Awasthi et al., 1984). The antiviral
agent was a basic glycoprotein (70-80% protein and 8-13% carbohydrates) with
a molecular weight of 16-20 kDa as determined by gel filtration chromatography
(Verma et al., 1979). The resistance-inducing
protein was found to be extermely thermostable (Verma and
Awasthi, 1979). Following treatment with the systemic resistance inducing
protein, the susceptible healthy host produces a virus inhibitory agent. The
glycoprotein occurring in B. diffusa roots functions as a signal molecule
and is of great interest as it has a role in stimulating the defense systems
of plants against viruses. Owing to the high antiviral efficacy of B. diffusa
under laboratory conditions, it was tested under field conditions as well against
a few viral diseases of economically important crops. The purified glycoprotein
from B. diffusa reduced infection and multiplication of tomato yellow
leaf curl virus papaya ring spot virus (Awasthi, 2000)
and cucumber green mottle mosaic virus (Awasthi et al.,
2003). The aqueous crude extract from the dried roots was also found significantly
active against a number of viruses mung bean yellow mosaic virus (Awasthi,
2000); bean common mosaic virus (Singh and Awasthi,
2002); bottle gourd mosaic virus in muskmelon (Cucumis melo), ridged
gourd (Luffa acutangula) and bottle gourd (Lagenaria siceraria) (Awasthi
and Kumar, 2003).
In treated plants, the antiviral agent from B. diffusa not only decreased disease symptom severity but also protected the plants against infection by viruses.
Root of Boerhaavia diffusa contains basal proteins which show high virus
inhibitory activity against plant viruses. Root extract of this plant induce
strong systemic resistance in susceptible host plant. In the study, we found
that the BD-SRIP induces the resistances against the TMV infection (Lohani
et al., 2007).
The aqueous extract of the leaves inhibited potato virus Y infection on chilli
plants (Suriachandraselvan and Narayanasamy, 1987).
Nitric oxide scavenging activity: The extracts of various polyherbal
drugs exhibited dose-dependent NO scavenging activities and the potency was
in the following order: Abana > Chyavanaprasha > Triphala>
Geriforte > Septilin> Mentat > Gingko biloba.
The present results suggest that the traditional Indian polyherbal crude drugs
may be potent and novel therapeutic agents for scavenging of NO and thereby
inhibit the pathological conditions caused by excessive generation of NO and
its oxidation product, peroxynitrite. These findings may also help to explain,
at least in part, the pharmacological activities like rejuvenating adaptogenic,
anti infection, anti-inflammatory, cardioprotective and neuroprotective activities
of these traditional, clinically used non toxic drugs because NO is an important
bioregulatory molecule which has a number of physiological effects including
control of blood pressure, neural signal transduction, platelet function. antimicrobial
and antitumor activity (Jagetia et al., 2004).
Adaptogen activity: Adaptogens seem to be useful during both adrenal
hyper stress as well as adrenal hypo fatigue. By definition, an adaptogen implies
the capability for bi directional or normalizing effects. The most important
adaptogens for the adrenals include Panax Ginseng, Siberian Ginseng, Ashwagandha,
Rhodiolia, Boerhaavia diffusa and Holybasil Leaf Extract. Boerhaavia
diffusa has the ability to support both adrenal over and under activation.
In stressful conditions it has demonstrated the ability to buffer the elevations
of serum cortisol and prevent the suppression of the immune system that takes
place with elevated cortisol. On the other hand, Boerhaavia diffusa has
also demonstrated the ability to improve cortisol levels with end stage adrenal
exhaustion (Mungantiwar et al., 1997).
Growth inhibition of struvite crystals: This in vitro study had
been carried out in the presence of herbal extract of Boerhaavia diffusa
Linn, by using single diffusion gel growth technique. Sodium metasilicate solution
of specific gravity 1.05 and an aqueous solution of ammonium dihydrogen phosphate
of 0.5 M concentration were mixed so that the pH value 7.0 could be set. After
the gelatin, equal amount of supernatant solution of 1.0 M magnesium acetate
prepared with 0.5 and 1% concentrations of the herbal extract of B. diffusa
Linn. were gently poured on the set gels in the respective test tubes in the
aseptic medium. The growth of crystals without and with herbal concentration
of B. diffusa Linn. increased the inhibition of crystals also increased
in the gel media as well as the dissolution of crystals at the gel-liquid interface
increases. The de-fragmentation of some grown crystals was also noticed (Vaidya
et al., 2009).
Anti fibrinolytic activity: A study that evaluates the effect of anti-fibrinolytic
Agents-Aminocapric Acid (-ACA). tranexamic acid (AMCA): anti-inflammatory drugs
(indomethacin. ibuprofen. naproxen); and plant extract (root extract of Boerhavia
diffusa) on endometrial histology of IUD-fitted menstruating monkeys. It is
effective in reducing stromal edema, inflammation, and tortuosity of glands,
and in increasing the degree of deposition of fibrin and platelets in the vessel
lumen (Barthwal and Srivastava, 1990).
Chemopreventive action: In the present study, cancer chemo preventive
property of B. diffusa was evaluated on 7,12-dimethyl Benz (a) anthracene
(DMBA) induced skin papillomagnesis in male swiss albino mice (6-7 weeks old).
This leads to the supposition that the inhibition of tumorigenesis by the plant
extract might have been executed either by preventing the formation of active
carcinogens from their precursors or by augmenting detoxification process, preventing
promotional events in the mouse skin through free radical scavenging mechanism
(Bharali et al., 2003).
Genetic diversity analysis: Boerhaavia diffusa is extensively
used in herbal medicines as well as in the Ayurvedic system, because it contains
a set of clinically important compounds. In the present study, the genetic variability
in Boerhaavia diffusa between accessions of different geographical origin
within the Indian Territory is assessed through random amplified polymorphic
DNA (RAPD) makers. Twenty-eight accessions of Boerhaavia were screened with
eighteen primers of which nine were found to be the most informative. The degree
of polymorphism was found to be high in accessions collected from different
places of Uttar Pradesh (Set II) in comparison to other states of India (Set
I). A relatively lower level of polymorphism was recorded in accessions collected
from diverse locations around Lucknow (Set III). Accessions from neighboring
geographical regions exhibited more similarity than those from distant regions
(as revealed by the set I analysis). Certain diagnostic makers may be correlated
with morphological character (s) such as plant type. BDL appeared most distinct
and divergent from the rest of the accessions and the BDJ plant in set II also
showed least similarity estimate. Fragments of 5.62 and 4.47 Kb with primer
GN59 was found to be unique for BDP and BD2 having ovate leaf character. Whereas
ovoid leaf genotype exhibited 0.79 Kb (GN34 primer) fragment. Similarly a unique
band type (0.35 Kb) with primer GN83 was present in BDL and BDI that share light
pink flower. Jaccard's and Nei and Li similarly coefficient values amongst the
accession were in the range of 0.22 to 0.89 and 0.33 to 0.93, respectively Association
of RAPD makers with the leaf characteristics, flower colour as well as with
geographical locations has been made. This shows that RAPD makers are also useful
for the study of genetic structure of Boerhaavia populations (Shukla
et al., 2003).
Bronchial asthma: Dried leaves are used in dhoomapana (smoking) in treatment
of bronchial asthma. The leaf decoction is an excellent expectorant when decocted
with punarnava (Boerhaavia diffusa) and then combined with ginger juice
and black pepper (Kala et al., 2009).
Antilymphoproliferative activity: It inhibited T cell mitogen phytohemagglutinin
and concanavalin A-stimulated proliferation of human Peripheral Blood Mononuclear
Cells (PBMC). It also inhibited purified protein derivative antigen-stimulated
PBMC proliferation and human mixed lymphocyte culture. In addition, B. diffusa
extract inhibited the growth of several cell lines of mouse and human origin,
such as mouse macrophage cells (RAW 264.7) human macrophage cells (U937), human
monocytic cells (THP-I), mouse fibroblast cells (L.929), human embryonic kidney
cells (HEK293), mouse liver cells (BNLCL.2), African green monkey kidney cells
(COS-I), mouse lymphoma cells (EL-4), human erythroleukemic cells (K562) and
human T cells (Jurkat). (Mehrotra et al., 2002b).
Antiproliferative and antiestrogenic activity: Treatment with varying
concentrations of BME (20-320 μg mL-1) resulted in moderate
to very strong growth inhabitation in MCF-7 cell lines. BME competed with [3H]-estradiol
for binding to ER with IC50 value of 320 25 g mL-1. RT-PCR
analysis revealed that BME reduced the mRNA expression of pS2 indicating the
ant estrogenic action of BME. BME treatment for 48 h resulted in a remarkable
increase in the number of MCF-7 cells in the Go-Gi fraction from 69.1 to 75.8%
with a reciprocal decrease of cells in all other phases indicating cell cycle
arrest at Go-Gi phase. Hence, it demonstrates that Boerhaavia diffusa
posses antiproliferative and Antiestrogenic properties and suggest that it may
have therapeutic potential in estrogen dependent breast cancers (Sreeja
and Sreeja, 2009).
Insecticidal activity: Chemical investigation of the root resulted in
isolation of insect moulting hormone which was structurally identified as-ecdysone.
Butanol extract of root was bioassayed on housefly (Musca domestica) last in
star larvae (Suri et al., 1982). The hexane and
acetone extracts of twigs showed insecticidal activity against Culex p. fatigans
and Musca domestica nebulo (Deshmukh et al.,
Toxicity of B. diffusa Linn.: Vomiting may be associated with larger doses of B. diffusa.
Plants contain thousands of constituents and are valuable sources of new and biologically active molecules possessing bioactivities. In spite of the tremendous strides in modern medicine, numerous natural products from traditional medicinal plants have been introduced in the development of theoretical drugs. In addition, many products containing herbal extracts are sold in the Asian market as substitutes or supplements of modern medicine. Recent years, ethno-botanical and traditional uses of natural compounds, especially of plant origin received much attention as they are well tested for their efficacy and generally believed to be safe for human use.
The objective of this review has been to show the recent advances in the exploration of plant Boerhaavia diffusa as phytotherapy and to illustrate its potential as a therapeutic agent. The available information in the literature on the bioactivities of the Boerhaavia diffusa shows that the plant contains compounds with strong pharmacological activities of potential clinical relevance and is a popular remedy among the various ethnic groups, Ayurvedic and traditional practitioners for treatment of various ailment. Researchers are exploring the therapeutic properties which are not known.
Authors are thankful to University Grant Commission, New Delhi, India for financial assistance.
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