Jadavpur

University
Dravyaguna
Vijnana

College : Bengal
Institute Of
Pharmaceutical
Sciences.
Name : Kuntal Mitra.
Roll No : 11.
 Class : 4th Yr 2nd sem.
2010.
Course : B.Pharm (Ayu).
Topic Bhumiamlaki
:
(Phyllanthus niruri).

Contents:
 Introduction
 Botanical Name
 Family
 Varnacular Name

 Synonims & Common Names

 Classical categorization
 Properties

 Habitat

 Indication,Therapeutic uses
 Botanical Description
 Pharmacological Uses

 Chemical constituents & Chemical

Structures

 Some medicines & Material and

Methods

 Research

 Conclusions

 References
 Dravyaguna :

Introduction:
 Phyllanthus niruri has a
long history in herbal medicine systems world wide. The whole
plant and its aerial parts are used for many remedies, mostly
biliary and urinary. Some examples are kidney and gallbladder
stones, hepatitis, colds, flu, tuberculosis, and other viral
infections. It has also been proven effective in liver diseases like
jaundice and liver cancer. It is sometimes used for bacterial
infections such as cystitis, prostatitis, venereal diseases, and
urinary tract infections. It can also assist in reducing anemia
symptoms, diabetes and hypertension. It is well known for its
diuretic, analgesic, stomachic, antispasmodic, febrifugal, and cell
protective properties. Phyllanthus may help decrease the amount
of hepatitis B virus found in the blood stream. Phyllanthus urinaria
is more effective medicinally than the variety Phyllanthusamarus.
Commonly known as stonebreaker because of
its strong roots, Phyllanthus Niruri is an herb that grows in tropical
areas like the rain forests of the Amazon, Bahamas, China and
Southern India. It can reach 30-60 centimeters in height and when
in bloom displays an array of yellow flowers. It grows freely like a
weed spreading abundantly. All parts of the plant exhibit
medicinal properties. It is used medically as a diuretic and an
astringent.In drier climates like Brazil, India, Florida, and Texas,
Phyllanthus amarus and Phyllanthus sellowianus grows. They are
similar in appearance and medical application to
PhyllanthusNiruri.
Phyllanthus
niruri, (family- Euphorbiaceae) is a herb found in many parts of
the world. It is known for a variety of uses viz. hepatoprotective
action, lipid lowering action, antidiabetic action, antifungal action
to name a few. It holds a reputed position in both Ayurvedic and
Unani systems of medicine.The article covers its phytochemical
constituents and major pharmacological activities.

Phyllanthus niruri
originated in India, usually occurring as a winter weed throughout
the hotter parts. The Phyllanthus genus contains over 600 species
of shrubs, trees and annual or biennial herbs distributed
throughout the tropical and subtropical areas. Phyllanthus niruri is
a herb of Euphorbiaceae family that grows upto 60 cm.
Phyllanthus means “leaf and flower” because the flower, as well
as the fruit, seem to become one with the leaf.
Phyllanthus niruri L.,
(Syn. P. fraternus Webster), Euphorbiaceae, is a common kharif
(rainy season) weed found in both cultivated fields and
wastelands. Recently it has attracted the attention of researchers,
because of its hepatoprotective properties. No effective specific
therapy is available for viral hepatitis but P. niruri has shown
clinical efficacy in viral Hepatitis B .It is known for its liver healing
properties so used in Chinese medicine for treatment of liver
diseases.
Bhumyamlaki (Phyllanthus niruri)

Botanical Name Phyllanthus niruri .

Family Euphorbiaceae.

Hindi : Bhuyiavla.
Varnacular
Bengali : Noar.
Name
Sanskrit : Bhoodatreo.

Tamil : Keelaneeli. Telegu : Nelausiri.

Kannada : Kirunelli , Konkani : Bhuin-avalae.

Uria : Narakoli , Malayalam : Kizhanelli.

Assamese : Holpholi. Mahrathi : Ray avali.

Tamalaki , Bhudhatri
Synonims Bahuphala , Bahupatra.
 Common
Phyllanthus Plant, Child Pick-aback, Gulf
Names Leafflower, Black Catnip, Meniran, Chanca
Piedra, Shatterstone, Stone Breaker, Quebra
Pedra, Gale Of Wind, Carry Me Seed,Creole
senna , Gale of wind , Carry Me seed.

Classical Caraka Kasahara ,

Svasahara
categorization
Susruta --------------
Rasa Tikya , Kasaya , Madhura.
Vagbhatta Kasahara ,
Guna Svasahara
Laghu , Ruksha

Virya Sita

Vipaka Madhura

Karma Kapha-pitta hara , Rechana.

 Properties :
 Habitat : Common in central and southern india extending to
Srilanka.

 Indication - Svasa-kasa , Prameha ,Rakta-pitta , Kamala .

 Therapeutic uses -
1) Kamala : Paste of bhuiamlaki may be taken with
butter milk.
2) Prameha : 2 gm of Bhuiamlaki & 20 maricha are
pounded together & taken may provide releif in
7 days.

 Part used - Whole plant , Root .

 Doses -Powder 3-5 gm , Paste 5-10 gm.
 Botanical Description -
The annual herb is 30-60 cm high, quite glabrous, stem
often branched at the base.

 Leaves : Numerous, sbsessile distichous often

imbricating, elliptic oblong obtuse. Stipules present,
very acute.
 Flowers : Yellowish, very numerous, axillary. The male
flowers are one to three in number while the female
flowers are solitary in nature.
 Capsules : 2.5mm in diameter, depressed globose,
smooth scarcelylobed.
 Pharmacological Uses –

I.Hepatoprotective Effect – 1.Hepatitis B is one of the major diseases

inflicting human population. Conventional treatment with
interferon – alpha is very expensive and has many serious side
effects. Alternative herbal medicine using extracts of Phyllanthus
niruri and Phyllanthus urinaria have been reported to be effective
against Hepatitis B and other viral infections. A study reports
quantitative determination of the anti viral effect of these herbs in
well-defined in vitro systems. 3
2.Phyllanthus niruri has been reported to exhibit marked
antihepatitis B virus surface antigen activity in in-vivo and in-
vitro studies. Infectious hepatitis is due to the inability of the
bodies’ immune system to eliminate the virus from the liver cells:
hence the “carrier state”. An infection with the virus is
documented by detectable levels of various viral antigens in the
blood, including HbaAg (the surface antigen of the virus) as well
as antibodies to the core of virus (HBc antibodies). In one study,
37 patients with chronic viral hepatitis B were treated with a daily
dose of 600mg of Phyllanthus niruri for 30 days. 59% of the
patients lost the HBsAg two weeks after the end of the treatment.
Furthermore, none of the cases followed for up to 9 months had
any symptoms of HBsAg. The authors postulated that Phyllanthus
niruri might inhibit proliferation of the virus by inhibiting
replication of the genetic material of the virus. 4
3. Hepatoprotective effect of an ayurvedic medicine; herbal
preparation HPN – 12 (containing Glycirrhiza glabr, Pichorhiza
kurroa, Berberis aristata, Piper longum, Phyllanthus niruri,
Solanum dulcamara, Zingiber officinale, Curculigo orchioides,
Elettaria cardamomum, Tinospora cordifolia, Desmodium trifolium
and Sacchrum officinarum) orally administered to male albino rats
at 1ml/100g body weight was found to be effective against liver
damage. 5
4. Animals with Carbon Tetrachloride induced hepatopathy were
treated with catliv (contains extracts of Swertia chirata, Eclipta
alba, Fumaria vaillanti, Picorrhiza kurroa, Andrographis paniculata
and Phyllanthus niruri) at 25ml twice daily orally for six days
starting at 48 hours after administration of Carbon tetrachloride.
On basis of result obtained it was concluded that the ingredients
in catliv, effectively helped in regeneration of hepatic cells and is
an effective liver tonic for calves.6
5. Research in Japan and India in the 1980's has demonstrated
the liver -healing properties of Phyllanthus niruri. The primary
compounds responsible are phyllanthin, hypophyllanthin and
triacontanal. Glycosides found in Phyllanthus niruri demonstrated
Aldose reductase (AR) inhibitory activity in studies conducted by a
Japanese research group in 1988 and 1989.7
II.HIV Replication Inhibition –

1.Aqueous extract of Phyllanthus niruri is reported to have

inhibitory effect on human immunodeficiency virus. The
investigation examines the anti-HIV effects of the alkaloidal
extract of Phyllanthus niruri in human cell lines. The inhibitory
effect on HIV replication was monitored in terms of inhibition of
virus induced cytopathogenecity in MT-4 cells. The alkaloidal
extract of Phyllanthus niruri showed suppressing activity on
strains of HIV-1 cells cultured on MT-4 cell lines. The CC50 for the
extract was found to be 279.85μgmL-1 whereas the EC50 was
found to be 20.98μgmL-1. Interestingly the Selectivity Index (SI)
was found to be 13.34, which showed a clear selective toxicity of
the extract for the viral cells. The alkaloidal extract of Phyllanthus
niruri was thus found to exhibit sensitive inhibitory response on
cytopathic effects induced by both the strains of human
immunodeficiency virus on human MT-4 cells in the tested
concentrations.8
2.Extracts of five medicinal plants: Aristolochia indica, Cassia
occidentalis, Phyllanthus niruri, Withania somnifera and Tinospora
cordifolia were administered to 10 HIV infected patients for a
period of six months to one year. The clinical status of the patient
and their CD4 cell counts were periodically monitored. The results
indicate that in seven of the ten patients, their CD4 count
increased and the patients remained either asymptomatic or their
clinical well being improved. There was no change in the CD4 cell
count in one of the patient and the other two progressed to full
blown AIDS.9

Natural liverprotecter

III. Lipid Lowering Activity -

1.Lipid lowering activity of Phyllanthus niruri alcoholic extracts in

triton induced hyperlipidaemia was examined in rats. It was
observed that administration of triton in rat caused increase in
serum cholesterol by 3.5 fold, phospholipid 2 fold and triglyceride
1.2 fold. Administration of Phyllanthus niruri at the dose of
200mg/kg simultaneously with triton lowered the level of total
cholesterol, phospholipid and triglyceride by 27, 25 and 24
percent respectively. In an experiment with cholesterol fed rats,
Phyllanthus niruri at a dose of 100 mg/kg lowered the elevated
level of low-density lipoprotein lipids in hyperlipidemic and drug
fed animals.10
IV.Anti – Diabetic Activity –

1.An alcoholic extract of Phyllanthus niruri was found to reduce

significantly the blood sugar in normal rats and in alloxan
diabetes rats. In normal rats, administration of Phyllanthus niruri
200mg/kg body weight reduced the blood sugar by 34.5 percent
and to 47.4 percent at the concentration of 1000mg/kg by weight
at 1 hour. However at 6th hour, values are almost similar to
normal value. Continuous administration of the drug produced
significant reduction in normal blood sugar in rats, which on 15th
day was also found to reduce the blood sugar in alloxan diabetic
rats. In short term experiment, drug was found to reduce the
blood sugar at 4th hour by 6.07 percent at dose level of 200mg/kg
by weight and 18.7 percent at concentration of 1000mg/kg by
weight. Continuous administration of drug produced significant
reduction in blood sugar in alloxan diabetic rats. On 15th day
values were almost similar to normal in the group taking
1000mg/kg by weight. Plant extract did not produce any toxicity
as seen from liver and kidney function test and in hematological
parameters. The results indicate potential antidiabetic action of
Phyllanthus niruri.11
V. Anti Malarial Activity –

1.The ethanolic, dichloromethane and lyophilized aqueous

extracts of Cassia occidentalis root bark, Morinda morindoides
leaves and whole plants of Phyllanthus niruri were evaluated for
their antimalarial activity in vivo, in 4-day, suppressive assays
against Plasmodium berghei ANKA in mice. No toxic effect or
mortality was observed in mice treated, orally, with any of the
extracts as a single dose, of 500 mg/kg body weight, or as the
same dose given twice weekly for 4 weeks (to give a total dose of
4 g/kg). No significant lesions were observed, by eye or during
histopathological examinations, in the hearts, lungs, spleens,
kidneys, livers, large intestines or brains of any mouse. At doses
of 200 mg/kg, all the ethanolic and dichloromethane extracts
produced significant chemosuppressions of parasitaemia (of >
60% for C.occidentalis root bark and Phyllanthus niruri whole
plant, and of 30% for M.morindoides leaves) when administered
orally. The most active ethanolic extract, that of Phyllanthus
niruri, reduced parasitaemia by 73%. The dichloromethane
extracts of M.morindoides and Phyllanthus niruri produced similar
reductions (74% and 72% chemosuppression, respectively),
whereas that of C.occidentalis was slightly less active (60%
chemosuppression). Each lyophilized aqueous extract was less
active than the corresponding ethanolic extract.12
VI.Activity Against Filarial Mosquito (Culex quinquefasciatus) –

1. 18 plants were evaluated for juvenile hormone analogue

activity against Culex quinquefasciatus. Of these acetone
extracts of 8 plants namely Commelina benghalensis , Ageratum
conyzoides , Achyranthus aspera, Sida acuta, Euphorbia
pulcherrina, Rivinia humilis, Ruellia tuberosa and Phyllanthus
niruri possessed significant juvenile hormone activity. The LC50
values of 5 most active plants namely Phyllanthus niruri,
Amaranthus spinosus, Antegonon leptopus, Corchorus aestuans,
Corchorus benghalensis were determined to be
13,16,17,17,14ppm respectively.13
VII.Anti- spasmodic activity –

1.Research done in Brazil at the Federal University of Santa

Catarina in 1984 on Phyllanthus niruri revealed an alkaloid
(phyllanthoside) in the leaves and stem with strong antispasmodic
activity. It served as a relaxing agent for smooth muscles and
they concluded that its spasmolytic action probably accounted for
the efficacy of Phyllanthus niruri in expelling stones.14
VIII. Analgesic activity –

1.Methanol extract of dried callus tissue at a concentration of

10mg/kg, administered intraperitonially to mice was active vs.
acetic acid induced writhin and vs. formalin – induced pedal
edema. The extract, at 50mg/kg was inactive vs tail flick response
to radiant heat. Ethanol/ water (1:1) extract of dried entire plant
at a dose of 50mg/kg, administered intragastric to male mice was
active. The extract also administered intraperitonially to male
mice at a dose of 0.3mg/kg was active. In both cases
antinociceptive effects were demonstrated using 5 different
models of nociception. 15
IX. Chromosome Aberration Inhibition –

1. Water extract of dried fruit and leaves, at a dose of 685.0

mg/kg, administered to mice by gastric incubation was active vs.
chromosome damage induced by lead nitrate and aluminium
sulphate in bone marrow chromosomes. Dosing was for 7 days.

 Worldwide Traditional Medicinal Uses-

 Bimini :

Hot water extract of the entire plant is administered orally, to

reduce fevers, and as a laxative.17

 Dominican Republic :

Hot water extract of leaves is administered orally as a popular

fever remedy.

 Fiji :

Decoction of dried leaves and roots is taken orally for fever, and
for good health. Dried entire plant, grounded in buttermilk is
administered orally for jaundice. Fresh leaf juice is used externally
for cuts and bruises. For eye diseases the juice is mixed with
castor oil and applied to the eye. Infusion of dried leaves is
administered orally for dysentery and diarrhea. Infusion of green
root is taken orally to treat heavy menstrual periods.19
 French Guyana :

Hot water extract of leaves is administered orally as a

cholagogue.

 Haiti :

Decoction of dried leaves is taken orally for or used in bath for

fever, and orally for indigestion.
Hot water extract of dried entire plant is administered orally as a
spasmolytic and is also against fever.
 India
Fresh plant juice is taken orally for genito urinary disorders .
The fruit is used externally for tubercular ulcers, scabies and
ringworm .
Hot water extract of dried entire plant is administered orally for
diabetes .
For asthma in ayurvedic medicine .
 Papau-New Guinea :

Fresh leaf juice or fresh root juice are taken orally for venereal
diseases. Decoction of dried entire plant is administered orally to
treat venereal diseases.
Decoction of dried leaf when taken orally is a treatment for
diarrhea. A cupful of leaf decoction is drunk daily.
 Philippines :

Decoction of dried entire plant is used as a bath for newborns. It

is believed to remove disease-causing elements from the skin.
Orally the decoction is used for coughs in infants.
 Puerto Rico :

Hot water extract of leaf and stem is taken orally for fevers .

 Tanzania :

Hot water extract of fresh entire plant is administered orally for

gonorrhea .

 Thailand :

Hot water extract of commercial sample of the entire plant, is

administered orally as an antipyretic .
Hot water extract of dried aerial parts administered orally is used
as a diuretic, as an antipyretic, and for malaria.
Hot water extract of dried entire plant is administered orally as
an anti-inflammatory agent.
 Virgin Islands :
Hot water extract of the plant is taken orally to increase the
appetite.

 West Indies :

Hot water extract of roots together with hot water extract of

Citrus aurantifolia roots is taken orally to increase appetite. Hot
water extract of entire plant administered orally, is taken for
malarial fever. The plant is boiled and the tea taken. Water
extract of the leaves and roots is taken orally for diabetes, and as
a diuretic .36
Phyllanthus niruri is used as a diuretic in dropsical affection,
gonorrhoea and other troubles of genito urinary tract. Herb is
bitter, astringent, diuretic and febrifuge, antiseptic. Fresh root is a
remedy for jaundice. Infusion of young shoots given in dysentery.
Leaves are popular remedy against fever. It can be used to
increase the appetite and locally to relieve inflammations. It can
also be used in case of anorexia.
x. In – Vitro Studies –

1.Role Of Calcium Oxalate Crystals – Alexander.H.Campos and

Nestor Schor investigated the in vitro effect of an aqueous
extract of Phyllanthus niruri L on the model of calcium oxalate
crystals endocytosis by Madin-Darby canine kidney cells; the
extract exhibited a potent and effective non concentration
dependent inhibitory effect on the calcium oxalate crystals
internalization. This response was present even at high
(pathologic) calcium oxalate concentration and no Phyllanthus
niruri L. –induced toxic effect could be detected. Biochemical
analysis of culture media containing Phyllanthus niruri L did not
provide any clues for the elucidation of the cellular pathways
affected by this natural product. Although further studies are
necessary for better understanding of the role of Phyllanthus
niruri L. in urolithiasis, their findings show that this natural
product could be an attractive alternative for the treatment of urinary
stone.

xi. Uses Of Isolated Phytochemical Constituents-

1.Bioassay – guided fractionation of boiled aqueous extracts from
the whole plant of Phyllanthus niruri (Euphorbiaceae) led to the
isolation of 1-o-galloyl-6-o-luteoyl-a-D-glucose (1), which IC50
values of 4.7mg/ml against Babesia gibsoni and 1.4mg/ml against
Plasmodium falciparum invitro. The known compounds b
glucogallin (2), quercetin 3-o-b-D-glucopyranosyl-(2 to 1)-o-b-
D- xylopyranoside(3), b-sitosterol and gallic acid were isolated.
Structures of these compounds were elucidated on the basis of
their chemical and spectroscopic data.

 General Features–
• Comparative pharmacognostic studies of 3 Phyllanthus
species –

The detailed pharmacognostic evaluation of the 3 species of

Phyllanthus namely Phyllanthus amarus Schum & Thonn or
Phyllanthus maderaspatensis Linn or mixture of Phyllanthus
amarus, Phyllanthus fraternus Webster and Phyllanthus
maderaspatensis has been carried out with the aim to establish
the identification markers of this important hepatoprotective
agents (effective in Hepatitis B too). The study concluded that the
3 species can be differentiated on the micro and macroscopic
characters, physicochemical values, HPTLC fingerprint profile and
the detection of phyllanthin and hypophyllanthin as marker
components. Besides, an interesting conclusion can be drawn that
phyllanthin and hypophyllanthin said to protect the hepatocytes
against CCl4 and galactose amine induced toxicity may not be
exclusively responsible for hepatoprotective activity as these are
present in Phyllanthus amarus while Phyllanthus fraternus and
Phyllanthus maderaspatensis also possess significant
hepatoprotective activity.

 Chemical Constituents & Chemical structures:

Rt – Rt culture , Pl – Plant , Lf – Leaf , Sd – Seed ,Ol – Oil , Aer –
Aerial plant , EO – Essential oils
1. Kaempferol-4-o-alpha-L-rhamnoside, Aer 0.9%, Rt
2. (-) Limonine, Lf EO 4.5% 3. Ascorbic acid, Lf 0.41%

4. Hypophyllanthin, Pl 0.05-0.17% 5. Geranin, Pl .23% ,7. Linoleic

acid, Sd Ol 21% ,8.Ricinoleic acid, Sd Ol 1.2% 9..Phyltetralin, Pl, Lf
0.14% ,10.&Phyllanthin, Lf, Aer .

 Following are parts of the plant : Ellagic acid, Pl

5. Estradiol, Pl
 6. Fisetin-41-O-beta-D-dlucoside, 7. Hinokinin, Pl , 8. Iso-
lintetralin, Pl , 9. Nirurin, Pl
10.Nirurinetin, Pl , 11.Phyllanthus, Pl , 12.Trans-phytol, Pl ,
13.Repandusinic acid A, Pl .
 Following are parts of the roots :
1. (+)-Catechin, Rt cult ,2. (+)-Gallocatechin, Rt Cult ,3. (-)-Epi-catechin, Rt
Cult
4. (-)-Epi-catechin-3-gallate, Rt Cult ,5. (-) Epi-gallocatechin, Rt
Cult .
Chemical Structures –

6. Gallic acid, Rt cult ,7. (-)-Epi-gallocatechin-3-O-gallate, Rt ,8.

Eriodictyol-7-o-alpha-L-rhamnoside, Rt 9.Fisetin-41-O-alpha-L-
rhamoniside, Rt .10.Lupeol acetate, Rt 11.Lupeol, Rt 12.Nor-
securinine, Rt .
 Following are parts of the Leaves :

1. 4-Hydroxy-lintetralin, Lf ,2. 2,3-dimethoxy-iso-lintertralin, Lf ,3.

Astragalin, Lf ,4. Beta sitosterol, Lf
5. Demethylenedioxy niranthin, Lf ,6. Hydroxy niranthin, Lf ,7.
Hypophyllanthin, Lf Aer ,8. Iso-quercitin, Lf
Chemical Structures –
 9. Linnanthin, Lf , 10.Lintetralin, Lf ,11. Niranthin, Lf ,
12.Quercitrin, Lf . 13 . Salicylic
acid methyl ester, Lf EO, 14. Seco-4-hydroxy-lintetralin, Lf.
 Following are parts of the aerial plant :

1.24- Isopropyl Cholestrol, Aer ,2.Dotriacontanoic acid, Aer ,

3.Nirphyllin, Aer ,4.Nirurine, Aer
5.Phyllanthenol, Aer, 6.Phyllantheol, Aer, 7.Phyllester, Aer ,
8.Phyllinurin, Aer ,9.Phylltetrin, Aer
10.Triacontan-1-al, Aer, 11.Triancontan-1-ol, Aer .

 Following are present in all leaf, stem, aerial plant, roots :

Chemical Structures –
1..Nirtetralin, Pl, Lf , 2.4-Methoxy-nor-securinine, Aer, Rt, St ,
3.Rutin, Pl, Lf 4. Phyllanthine, Rt, Lf, St , 5. Phyllochrysine, Lf, St ,
6. Quercetin, Lf, Pl.

 Some medicines made by Bhumyamlaki (Phyllanthus niruri)

 Material and Methods :

Plant Material :
The whole plant of Phyllanthus niruri
(Bhuiamlaki) was obtained from Amrutlal brothers Ltd. Mumbai.
M/s. Anushka Herbals, Mumbai, India, performed the identification
and authentication of the plant and a voucher specimen of
Phyllanthus niruri has been deposited with them. The plant was
then ground into a mixer to obtain the coarse powder.
Plant Extraction : (Alkaloidal Fraction) :
The dried, coarsely powdered whole plant of
Phyllanthus niruri (200g) was moistened with 25% ammonium
hydroxide, allowed overnight standing and then Soxhlet extracted
with 95% ethanol. After concentration under vacuum, the syrupy
residue (30g) was treated with concentrated hydrochloric acid.
The acidic filtrate was washed with benzene, made basic (pH 10)
with 25% ammonium hydroxide and extracted with chloroform to
afford the alkaloidal fraction (2g).
Chemicals and Reagents :
The chemicals/reagents used included RPMI
1640 medium (Gibco, grand Island, NY, USA), Foetal Calf Serum
(FCS) (Whittakar Bioproduct, MD, USA), 3-(4,5 Dimenthylthiazole-
2-yl) 2,5-Diphenvltetrazolium bromide (MTT) (Wako Pure
Chemical, Osaka, Japan), Fluorescin isothiocyanate-Conjugated
F(ab') 2 fragments of rabbit anti-human immunoglobulin antibody
(Cappel, Organon Taknika Corp., West Chester, PA, USA).
Culture of T-Cells :
The HTLV-1 infected MT-4 Cell live was
established by co-cultivating leukocytes from an adult T-cell
leukemia (ATL) patient with cord blood Leukocytes (CBL) (Miyoshi
et al 1981).
All cell lines were grown and maintained under the same
conditions as the H9/HTLV-IIIB cell line.
Human anti-HIV serum :
A serum with high titre of antibodies against
HIV was obtained from a patient. The serum was found positive by
ELISA-HTLV-III (ratio> 13.5), Western blot analysis and indirect
immunofluorescence in lymphadenopathy associated Molt cells
(titre of 1:1000). A dilution in PBS of the positive serum (stored in
aliquots at -20°C) was used in all experiments.

 Research :
 The effect of Phyllanthus niruri on urinary inhibitors of calcium
oxalate crystallization and other factors associated with renal
stone formation :

 ABSTRACT
Objective- To evaluate the effect of an aqueous extract of Phyllanthus niruri (Pn), a
plant used in folk medicine to treat lithiasis, on the urinary excretion of endogenous
inhibitors of lithogenesis, citrate, magnesium and glycosaminoglycans (GAGs).

Materials and methods -The effect of chronic (42 days) administration of Pn

(1.25 mg/mL/day, orally) was evaluated in a rat model of urolithiasis induced by the
introduction of a calcium oxalate (CaOx) seed into the bladder of adult male Wistar rats.
The animals were divided into four groups: a sham control (16 rats); a control+Pn (six);
CaOx+water instead of Pn (14); and CaOx+Pn (22). Plasma and urine were collected
after 42 days of treatment for biochemical analysis and the determination of urinary
excretion of citrate, magnesium and GAGs. The animals were then killed and the calculi
analysed.

Results -The creatinine clearance or urinary and plasma concentrations of Na+, K+,
Ca2+, oxalate, phosphate and uric acid were unaffected by Pn or the induction of
lithiasis. Treatment with Pn strongly inhibited the growth of the matrix calculus and
reduced the number of stone satellites compared with the group receiving
water. The calculi were eliminated or dissolved in some treated animals
(three of 22). The urinary excretion of citrate and magnesium was
unaffected by Pn treatment. However, the mean (SD) urinary concentration
of GAGs was significantly lower in rats treated with CaOx+Pn, at 5.64
(0.86) mg/g creatinine, than when treated with CaOx + water, at
11.78 (2.21) mg/g creatinine. In contrast, the content of GAGs in the calculi was higher
in the CaOx + Pn rats, at 48.0 (10.4) g/g calculus, than in the CaOx + water group, at
16.6 (9.6) g/g calculus.

Conclusion - These results show that Pn has an inhibitory effect on crystal growth,
which is independent of changes in the urinary excretion of citrate and Mg, but might be
related to the higher incorporation of GAGs into the calculi.

• AIDS Research and Human Retroviruses :

 HIV-1 Reverse Transcriptase Inhibitor from Phyllanthus
niruri:
TAKAHIRO OGATA,1
HIROTAKA HIGUCHI,1
SHUNJI MOCHIDA,1
HIDEKI MATSUMOTO,1
AKIHISA KATO,2
TOHRU ENDO,1
AKIRA KAJI,3
HIDEKO KAJI2

An aqueous extract of Phyllanthus niruri

(Euphorbiaceae) inhibited human immunodeficiency virus type-1
reverse transcriptase (HIV-1-RT). The inhibitor against HIV-1-RT in
this plant was purified by combination of three column
chromatographies, Sephadex LH-20, cellulose, and reverse-phase
high-performance liquid chromatography. The inhibitor was then
identified by nuclear magnetic resonance (NMR) spectra as
repandusinic acid A monosodium salt (RA) which was originally
isolated from Mallotus repandus. The 50% inhibitory doses (ID50)
of RA on HIV-1-RT and DNA polymerase α(from HeLa cells) were
0.05 μM and 0.6 μM, respectively, representing approximately a
10-fold more sensitivity of HIV-1-RT compared with DNA
polymerase α. RA was shown to be a competitive inhibitor with
respect to the template-primer while it was a noncompetitive
inhibitor with respect to the substrate. RA as low as 10.1 μM
inhibited HIV-1-induced cytopathogenicity in MT-4 cells. In
addition, 4.5 μM of RA inhibited HIV-1-induced giant cell formation
of SUP-T1 approximately 50%. RA (2.5 μM) inhibited up to 90% of
HIV-1 specific p24 antigen production in a Clone H9 cell system.

• Lipid lowering activity of Phyllanthus niruri in hyperlipemic

rats :
 Abstract : The lipid lowering activity (LLA) of Phyllanthus niruri
has been studied in triton and cholesterol fed hyperlipemic
rats. Serum lipids were lowered by P. niruri extract orally fed
(250 mg/kg b.w.) to the triton WR-1339 induced hyperlipemic
rats. Chronic feeding of this drugs (100 mg/kg b.w.) in animals
simultaneously fed with cholesterol (25 mg/kg b.w.) for 30
days caused lowering in the lipids and apoprotein levels of
VLDL and LDL in experimental animals. The LLA of this drug is
mediated through inhibition of hepatic cholesterol biosynthesis, increased
faecal bile acids excretion and enhanced plasma lecithin: cholesterol
acyltransferase activity.
 Abbreviations : HDL, high density lipoprotein; LCAT,
lecithin: cholesterol acyltransferase; LDL, low density
lipoprotein; LLA, lipid lowering activity; LPL, lipoprotein lipase;
PL, phospholipids; TC, total cholesterol; TG, triglyceride; TIHR,
triton induced hyperlipemic rats; VLDL, very low density
lipoprotein
 Effect of Extract of Phyllanthus niruri on crystal
deposition experimental urolithiasis :
Phyllanthus niruri (Pn) is a
plant that has been shown to interfere in the growth and
aggregation of calcium oxalate (CaOx) crystals. In the present
study we evaluated the effect of Pn on the preformed calculus
induced by introduction of a CaOx seed into the bladder of male
Wistar rats. Pn treatment (5 mg/ rat/day) was initiated
immediately or 30 days after CaOx seeding and thus in the
presence of a preformed calculus. Animals were sacrificed 50 or
70 days after surgery. The resulting calculi were weighed and
analyzed by X-ray diffraction, stereomicroscopy and scanning
electronic microscopy. Precocious Pn treatment reduced the
number (75%, P < 0.05) and the weight (65%, P < 0.05) of calculi
that frequently exhibited a matrix-like material on its surface,
compared to the untreated CaOx group. In contrast, Pn treatment
in the presence of a preformed calculus did not prevent further
calculus growth; rather, it caused an impressive modification in its
appearance and texture. Calculi from Pn-treated animals had a
smoother, homogeneous surface compared to the spicule shape
of calculi found in the untreated CaOx group. XRD analysis
revealed the precipitation of struvite crystals over the CaOx seed
and Pn did not change the crystalline composition of the calculi.
This suggests that Pn interfered with the arrangement of the
precipitating crystals, probably by modifying the crystal-crystal
and/or crystal-matrix interactions. Results suggest that Pn may
have a therapeutic potential, since it was able to modify the
shape and texture of calculi to a smoother and probably more
fragile form, which could contribute to elimination and/or
dissolution of calculi.

Conclusions–
1.Many times Phyllanthus niruri is adulterated with Phyllanthus
amarus and vice versa.
2.Many researchers have concluded that Phyllanthus niruri and
Phyllanthus amarus are same due to their similar chemical
constituents and pharmacological action. But it is not so as they
belong to different species
3.Phyllanthus niruri has variegated uses like hepatoprotective
effect, inhibition of HIV replication, lipid lowering activity, anti
diabetic activity, anti malarial, antifungal, antispasmodial activity,
etc.
4.Due to all these applications we also get Phyllanthus niruri as
marketed preparation by some prominent companies. In the
marketed preparations Phyllanthus niruri is added in everyday
used medicines like chyawanprash of many prominent
companies. Phyllanthus niruri powder is also available for use.