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2.1
Introduction
Numerous disease prevailing in the society can be treated using
medicines. Various parts of plants such as leaf, stem, roots and barks
were used as medicines in auyrveda, siddha, unani and homeopathic
treatments.
In Ayurveda: The Argemone mexicana linn. is useful in guineaworm infestations, purgative and diuretic. Seeds of the plant are used as
an antidote in snake poisoning and also acts as an emetic, expectorant,
demulcent and laxative. The protein-dissolving substances containing
seed extract is used to cure warts, cold sores, cutaneous infections,
itches, jaundice and dropsy 1. Seeds are effective against skin infection,
sores, dropsy and jaundice 2. Juice of the plant cures ophthalmic and
opacity of cornea. Oil of the seed is used to treat skin diseases. Roots are
antihelmentic
and
also
used
in,
skin
diseases,
leprosy
and
inflammations3.
In Homeopathic medicine: The drug prepared from this plant is
very useful in treating the problem caused by tape worm. The whole
plant is reported to be used for the treatment of whooping cough and
bronchitis 4.
In Unani medicine: Argemone Mexicana linn. helps in the
enrichment of blood which acts as an expectorant and aphrodisiac. It is
also used in treating skin diseases and leucoderma 5.
In siddha medicine: Argemone Mexicana linn. is widely used to
cure venereal sores, scorpion bite,photophobia, leucorrhoea. Leaves
along with black pepper are used to cure diabetes. The latex of A.
mexicana used to treat boils by topical application on the site of boils.
Whole plant is used to treat dental disorders. Leaf decoction is used in
the treatment of malarial fever and ulcers. Juice of the plant is applied
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on scorpion sting. Seeds are effective against leprosy, jaundice and
dropsy 6.
2.2
Antimicrobial activity
Acetone, ethyl acetate and petroleum ether extracts of leaf and
stem of Argemone Mexicana linn. was tested against as E. coli, Shigella
sp., Staphylococcus sp. and Salmonella sp. for their antibacterial and
antifungal properties. Excellent activity was exhibited by petroleum
ether extract of both leaf and stem, ethyl acetate extract for both leaf and
stem showed moderate activity and the acetone extracts were found
inactive against these water borne pathogens 7.
Singh et al in 2009 have reported antibacterial property of
different seed extracts viz. n-hexane, chloroform, methanol and water. It
was found that chloroform extract exhibited excellent results against all
the bacterial and fungal strains along with multidrug resistant P.
aeruginosa and S.aureus. It was found that methanol extract exhibited
moderate activity against all the microbial strains. However, no activity
was reported against all the pathogens for n-hexane and aqueous
extracts8.
Crude extracts of methanol and cold water of both leaves and
seeds of Argemone mexicana linn. were tested against multi drug
resistant pathogenic bacterial strains. The activity was carried our using
agar plate diffusion method. Both the extracts were found to exhibit
excellent activity9.
Argemone mexicana linn. oil extracted from bark and root
exhibited positive acivity against S. aureus, B. subtilis, K. pneumoniae,
P. aeruginosa, C. albicans, C. stellatoidea and C. torulopsis. It posses
efficacy in inhibition of growth of selected bacteria and fungi among the
tested organism.10
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N-dimethyloxysanguinarine isolated from chloroform extract of
A. mexicana showed antibacterial activity against K. pneumonia, S.
aureus, E. coli and P. aeroginosa with minimum inhibitory
concentration of 1.5625 to 3.125 11.
The extract of alcohol and petroleum ether from aerial parts of
Argemone mexicana linn. were evaluated for antimicrobial activity
against two bacterial strains (gram positive and negative), Bacillus
subtilis and Escherichia coli. Both extracts showed very good
antimicrobial activity on the pathogens. Comparatively alcohol extract
showed higher activity than petroleum ether extract 12.
2.3
Antidiabetic activity
Rout et al in 2011 reported hypoglycemic potentiality of aerial
parts of Argemone mexicana linn. in Alloxan induced diabetic rats.
Treatment was given for 11 days with ethanol and aqueous extracts at a
dose of 200 and 400 mg/kg body weight in diabetic rats. Treatment of
aqueous extract (400 mg/kg body weight) exhibited significant reduction
in blood glucose levels, triglyceride, cholesterol, plasma urea, creatinine
values and recovery in body weight compared to diabetic control rats
and the standard drug treated rats 13.
Ariel parts of the plant Argemone mexicana linn. were extracted
in water-alcohol mixture (200 and 400 mg/kg body weight), resulted in
reducing fasting blood glucose levels in Streptozotocin induced
hyperglycemic wistar albino rats14.
2.4
Antioxidant activity
Different extracts of Argemone mexicana linn. leaves were found
to possess super oxide anion scavenging activity by nitro blue
tetrazolium assay. Even at the dosage of 200 μg/ml; all the extracts
exhibited maximum percentage of antioxidant property. The highest
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radical scavenging activity was exhibited by acetone extract whose IC50
value was double to that of L-ascorbic acid value15.
Ethanolic extract of Argemone mexicana linn. root is reported to
show high DPPH radical scavenging activity (85.17%), ABTS radical
scavenging activity (75.27%) and H2O2 radical scavenging activity
(84.25%) at 100 μg/ml concentration. Extract showed dose dependent
high free radical scavenging activity 16.
2.5
Anti malarial activity
Malaria was treated with a potential, dose-escalating, quasiexperimental clinical trial method with a traditional healer using a
decoction of Argemone mexicana linn.. The remedy was prescribed in
three regimens: once daily for 3 days to group A; twice daily for 7 days
to group B; and four times daily for the first 4 days followed by twice
daily for 3 days to group C. All presented to the traditional healer with
symptoms of malaria and had a Plasmodium falciparum parasitaemia
>2000/μl but no signs of severe malaria. The proportions of adequate
clinical response (ACR) at Day 14 were 35%, 73% and 65% in Groups
A, B and C, respectively (P = 0.011). At Day 14, overall proportions of
ACR were lower in children aged <1 year (45%) and higher in patients
aged >5 years (81%) (P = 0.027). Very few patients had complete
parasite clearance, but at Day 14, 67% of patients with ACR had a
parasitaemia <2000/μl17.
2.6
Wound Healing activity
Different solvent extract of Ar gemone mexicana linn. leaves
were examined for wound healing activity in wistar albino rats. The
activity was carried out by excision, incision and dead space wound
models. The results showed that the treatment with methanol extract of
leaves of Argemone mexicana linn. accelerated wound healing agent in
rats18.
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The wound healing effects of the leaf extract (50% ethanol) and
latex of Argemone mexicana linn. were investigated on albino rats using
both excision and incision wound models. Topical application of the
extract and latex, respectively, gave 67.08 and 57.86% healing after 12
days in the excision model and increased tensile strength to 188.50 and
154.61 gm in the incision model. However, both the extract and latex of
Argemone mexicana linn. were not as effective as the standard
nitrofurazone in both excision and incision models 19.
2.7
Hepatoprotective activity
Extracts of root bark of Argemone mexicana linn. were
investigated on CCl4 (Carbon tetra Chloride) induced liver damaged in
rats. Acute toxicity study, efficacy study, blood and tissue biochemical
assays like ALT, AST, bilirubin, total protein, glucose, LM and EM etc
have been studied for evaluation of Hepatoprotective action. From
above observations of biochemical parameters it was observed that
Argemone mexicana linn. has a high potential in healing liver
parenchyma and regeneration of liver cells hence it may act as a liver
tonic with high potency20.
The protective effects of the aqueous extract of Argemone
mexicana linn. whole plant; against CCl4 induced hepatic failure in male
albino rats (Wistar strain) was investigated. For acute and massive
invasion of hepatopathy, CCl4 (i.p injection of CCl4+Olive Oil in 1:1
ratio; 2ml/kg) was used and various biochemical parameters followed by
significant (p<0.001) weight loss in toxic control group (-12.83±1.13).
The administration of aqueous extracts (250mg/kg and 150mg/kg of
body weight) for 7 days, elicited protective action since the elevated
levels of marker enzymes (AST, ALT, ALP) of liver functions were
found to be decreasing progressively in a dose dependent manner with
net weight gain. In the aqueous extract, 250mg/kg treated rat group all
the marker enzymes were analysed to be decreasing significantly
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(p<0.001), (AST, 272.77±24.08; ALT, 189.15±7.16; ALP, 97.15±6.54)
and the final body weight was also significantly (p<0.001) increased
(6.16±1.01) when compared with the toxic control group. The serum
total protein and the serum albumin were also approaching normal
values. The results found in aqueous extract 250mg/kg treated rat were
quite promising and were comparable with a standard polyherbal drug
Liv-52. The statistically processed results support the conclusion, that
the aqueous extract of Argemone mexicana linn. whole plant (250mg/kg
and 150mg/kg) possesses dose dependent, significant protective activity
against CCl4 induced hepatotoxicity 21.
2.8
Antihelmintic activity
Alcoholic and aqueous extract of leaves of Argemone mexicana
linn. were tested for antihelmintic activity in a dose dependent manner
(6.25, 12.5, 25, 50, 100 mg/ml) against Pheretima posthuma and
Ascardia galli. Both the extracts were found to posses significant
antihelmintic activity at 100 mg/ml concentration 22.
2.9
Antiplasmodial activity
Among the different species exhibiting low IC50 value between 943 mg (dry extract) / ml pertaining to posses good antiplasmodium
property, Argemone mexicana linn. (Papavaraceae) also showed the
same activity which varied by varying extract dose per kg/body wt.
concentration: In vitro inhibition (%) of plant extracts against
chloroquine susceptible strain of P. falciparum. The result showed mg
dry extract / 80 mg part dry plant material 2.50, % Inhibition (mg:ml)
according to dose i.e. 100 μg/ml (87); 50 μg/ml (76); 25 μg /ml (60);
12.5 μg /ml (51); and IC50 μg /ml 23.
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2.10
Larvicidal activity
Acetone fraction of petroleum ether extract of Argemone
mexicana linn. seed was reported to possess larvicidal activity. Extract
resulted in 100% mortality of the larva with LC50 value of 13.58 and
17.43 in field and laboratory conditions respectively 24.
Acetone fraction of the petroleum ether extract of seeds of
Argemone mexicana linn. exhibited larvicidal and growth inhibiting
activity against the second instar larvae of Aedes aegypti25.
The leaf extract (in petroleum ether) of the plant also exhibits
high larvicidal potential with LC50 value of 48.89 ppm against 3rd -4th
instar larvae of Culex quinquefasciatu26.
2.11
Anticancer activity
The ethanol extract of Argemone mexicana linn. was reported to
exhibit inhibitory activity against human cancer cell lines such as HelaB75 (48%), HL-60 (20.15%) and PN-15 (58.11%) 27.
2.12
Vasorelaxant activity
The vascular effects of a methanolic extract of the aerial part
Argemone mexicana linn. was investigated in rat aormatic rings. The
extract produced relaxation from contraction induced by norepinephrine
(NE: 1X10-7 M) in a concentration-dependent manner (0.1 at 1000
μg/ml). At higher concentrations (310 and 1000 μg/ml), the extract
induced significant additional tension. When applied to relaxed aortic
tissue, addition of the extract produced concentration dependent
contraction.
Vasoconstrictor and vasorelaxant effects produced by the extract,
were not altered significantly in endothelium-denuded rings, however,
phenoxi- benzamine (1 mg/ml) produced a rightward shift in the
concentration–contractile response curve to the extract. On the other
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hand, the extract relaxed significantly the contraction induced by KCl
(40 mM). The results indicate that the extract induces a direct dual
specific effect upon the vascular smooth muscle, mediated, at least in
part, by adrenergic receptors 28.
Petroleum ether, CHCl3, CHCl3/MeOH (9:1), MeOH extract and
partially purified fractions and pure compounds from Argemone
mexicana linn. examined on the electrically induced contractions of the
isolated guinea-pig ileum (E.C.I.). The results of the experiments
indicate that CHCl3/MeOH (9:1) and MeOH extracts, tested at a
concentration of 400, 200 and 100 mg/mL, dose-dependently reduced
the guinea-pig ileum contractions, whereas the petroleum ether and
CHCl3 extracts did not affect it.
Furthermore, the partially purified fractions II-V from the MeOH
extract, each tested at concentrations of 200, 100 and 50 mg/mL also
inhibited E.C.I. Finally the pure compounds 1-2 isolated and purified
from the above fractions significantly reduced, in a dose dependent
manner, the electrical contractions of the ileum, whereas compound 3
increased them. Since the active compounds 1-3 were respectively
identified as protopine, allocryptopine and berberine by NMR, the
observed effects could be related mainly to these compounds 29.
2.13
Antiasthmatic activity
Manuscript published in 2007 by Rao et al showed the
significance of Argemone mexicana linn. (Papaveraceae) (common or
local name: Brahmadand) Seed powder as antiasthamatic if taken 100–
200 mg twice a day, for 2 weeks 30.
2.14
Cytotoxic activity
The alkaloid isolated by fractionation of the chloroform extract
from the aerial part of Argemone Mexicana linn. was evaluated for its
cytotoxicity to human nasopharyngeal carcinoma (HONE-1) and human
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Review of Literature
gastric cancer (NUGC) cell lines. Chelerythrine was found to exhibit
significant activity against NUGC cell line, while angoline inhibited
both types of cell lines. (+)-Argenaxine showed moderate activity
against the NUGC cell line 31.
2.15
Anti-HIV activity
Anti-HIV activity was conducted for the alkaloids extracted from
the methanolic extracts of the Argemone mexicana linn. In which
benzo[c]phenanthridine (+/-)-6-acetonyldihydro chelerythrine exhibited
significant anti-HIV activity in H9 lymphocytes with EC50 and TI
(Therapeutic Index) values of 1.77 μ /ml and 14.6, respectively 32.
2.16
Molluscicidal activity
Molluscicidal property of seeds of Argemone mexicana linn.
against snail Lymnaea acuminata was studied. It was observed that
molluscicidal activity of seed powder of Argemone Mexicana linn., was
both time and dose dependent. Protopine and sanguinarine in seed of
Argemone mexicana linn. was identified as the active moiety causing
snail death by co-migration of active agent with seed powder 33.
2.17
Neuropharmacological studies
The ethyl acetate and methanol extract of the whole plant of
Argemone mexicana linn. was reported for analgesic activity, locomotor
activity and muscle relaxant activity using Wistar albino mice at an oral
dosage of 100, 200 and 400 mg/kg body weight. Both extracts showed
significant activities but methanol extract at a dosage of 200 mg/kg body
weight was found to be more potent for central nervous system activities
such as analgesic, anxiolytic and sedative effects 34.
2.18
Toxicity and safety evaluation
The alkaloid sanguinarine isolated from seeds of Argemone
mexicana linn. was examined for its hepatotoxic potential in rats. The
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studies exhibited that a single i.p. dose (10 mg/kg) of sanguinarine not
only increased the activity of SGPT and SGOT substantially but also
caused a significant loss of microsomal cytochrome P-450 and
benzphetamine N-methylase activity. Furthermore, the treated rats
exhibited considerable loss of body and liver weight, peritoneal edema
and slightly enlarged livers with fibrinous material. Microscopic
examination of the liver tissue showed progressive cellular degeneration
and necrosis further substantiating that sanguinarine is a potential
hepatotoxic alkaloid 35.
Toxicolethal effects of seeds of Argemone Mexicana linn. were
investigated in to roof rat, (Rattus rattus L). The argemone seeds were
fed at 100% of the diet up to the death or for a maximum of 10 days.
Observed signs of poisoning were sedation, passiveness, sluggishness,
feeble or no muscular jerks, abdominal contractions and increased
defecation. Also black secretions from the eyes, corneal opacity,
erection of hairs, and edema of the hind legs and submandibular space in
were noted. Fourteen of sixteen rats died. Significant reduction in the
weights of the rats was observed. There were significant increases in
blood glucose, BUN and SGOT. Major histopathological lesions were:
hepatocytolysis, nuclear degeneration, pyknosis, cloudy swelling and
dilatated sinusoids disturbing the lobulalar architecture of the liver;
proliferated endothelium of glomeruli, hemorrhage in glomeruli and
interstitium, and cloudy swelling of convoluted tubular epithelium in the
kidney cortical region; erosion and atrophy of the upper stomach
mucosa and calcification in the cardiac stomach, and; erosion and
congestion of the upper mucosa of the duodenum. No change was
noticed in the ileum 36.
Safety evaluation studies on argemone oil through dietary
exposure for 90 days in rats: Epidemic dropsy is a disease caused by the
consumption of mustard oil contaminated with argemone oil (AO).
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During 1998 dropsy in New Delhi, which is so far the largest with more
than 3000 victims and over 60 deaths, it was enquired at various
scientific and regulatory meetings about the maximum tolerated dose of
AO. Animals were given AO in diet at a dose of 0.001%, 0.01%, 0.1%,
0.5% and 1% daily for 90 days and the two control groups received the
standard diet with and without 1% mustard oil. A decrease in body
weight gain (28–31%) was observed in 0.5% and 1% AO groups; while
significant increases in relative lungs and liver weight was noticed in
respective doses of 0.01% and 0.1% AO groups as well as in higher
dosage animals. Reduction in RBC count and haemoglobin content (p <
0.05) was noticed in 0.01% and 0.1% AO exposed animals. This effect
was more pronounced in higher AO doses. Serum marker enzymes
including alanine transaminase (ALT), aspartate transaminase (AST),
lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) were
found to be significantly elevated in 0.01–1% AO groups.
Further, a decrease in albumin/globulin ratio (42–78%) was
observed in the serum of 0.01% to higher AO dose groups. The levels of
serum triglycerides and VLDL cholesterol were found to be enhanced (p
< 0.05) in AO treated (0.01–1.0%) animals. Histopathological changes
in lung were observed at 0.01% dose of AO while liver, kidney and
heart produced changes at 0.1% AO and above doses.
None of the parameters were found to be affected in 0.001% AO
treated animals. These results suggest that the no observed adverse
effect level (NOAEL) dose of AO is 0.001% in rats and considering a
factor of 100 for humans for highly toxic compound, the safe limit of
0.00001% (100 ppb or 100 ng AO/g oil) AO can be implicated which
shall contain only 0.55% of sanguinarine equivalent to 0.6 ng
sanguinarine per gram oil. However, the minimum detectable limit of
AO is 5 ppm (equivalent to 5 ng sanguinarine per gram oil) with the
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present existing HPLC method, thereby suggesting that mustard oil
should be absolutely free from AO contamination 37.
In vivo DNA damaging potential of sanguinarine alkaloid,
isolated from argemone oil, using alkaline Comet assay in mice:
Consumption of mustard oil contaminated with argemone oil is well
known to cause clinical manifestation referred to as ‘‘Epidemic
Dropsy’’. Since, sanguinarine alkaloid is the major component of
argemone oil, the in vivo DNA damaging potential of the isolated
alkaloid was investigated in blood and bone marrow cells of mice using
alkaline Comet assay. Swiss albino male mice were given single
intraperitoneal administration of 1.35, 2.70, 5.40, 10.80 and 21.60 mg
sanguinarine alkaloid/kg bwt., while controls were treated with saline in
the same manner. The results revealed a dose dependent increase in
DNA damage in blood and bone marrow cells following 24h treatment
of sanguinarine alkaloid. All the three parameters of Comet assay
including olive tail moment (OTM), tail length and tail DNA showed
significant (p < 0.05) increases in blood and bone marrow cells at
respective doses of 10.80 and 5.40 mg alkaloid/kg . However, some of
the parameters were significantly increased even at lower doses of
sanguinarine alkaloid (2.70 mg/kg).
The frequency of cells exhibiting greater DNA damage was
found to be increased by sanguinarine alkaloid in a concentration
dependent manner. These results indicate that single exposure of
sanguinarine alkaloid causes DNA damage in blood and bone marrow
cells of mice, which could be responsible for the genotoxicity of
argemone oil. The present study clearly indicates that sanguinarine
alkaloid, an active ingredient of argemone oil possesses DNA damaging
potential in blood and bone marrow cells using alkaline Comet assay.
These results fully support the earlier observation that in vivo argemone
oil caused genotoxicity by enhancing the frequencies of chromosomal
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aberrations, micronuclei formation and development of Comets resulting
in DNA damage34. In this regard studies have shown that sanguinarine
forms DNA adducts following metabolism by cytochrome P-450 system
under in vitro conditions 38.
It has been suggested that sanguinarine may undergo Ndemethylation by cytochrome P-450. Since, sanguinarine has been
shown to cause inactivation of cytochrome P-450, it can be argued that
the N-demethylated product of sanguinarine or any other electrophilic
metabolite, could be responsible for this effect. The decrease in
cytochrome P-450 thereby impairs the elimination of a metabolite of
sanguinarine, identified as benzacridine, in urine and feces. Although,
minimum group of sanguinarine has been shown to have affinity with bform duplex DNA by intercalation with a high preference to GC base
pairs nonetheless, it could not reveal genotoxicity in SOS chromtest
using E. coli PQ37 in the absence and presence of metabolic activation
system. However, it raised the possibility of usage of Sanguinaria
extract in toothpaste in the development of oral leukoplakia 39.
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