Used in Unan; S,S'•• 01" I .I- - lul"n Shakir Jamil*, Shoaib Ahmad**, Jamal Akhtar and Khursheed Alam Dept. of Moalijat, Faculty of Unani Medicine, lamia Hamdard, New Delhi-l10062, India * Correspondent author, E-mail: [email protected]**1437, Sector 39 - B, Chandigarh-160036, India Introduction Diseases caused by protozoa are responsible for a considerable morbidity and mortality,especially in the developing world. Amoebic dysentery or amoebiasis is one of such protozoal diseases. It is caused by Entamoeba histolytica. If untreated, it may give rise to serious complications including liver abscesses. There are some 42 million cases annually and an estimated 75,000 deaths across the world due to this disease. In allopathic system ofmedicine the disease is generally treated with metronidazole, although this drug is poorly tolerated by some patients. There are a number of antiamoebic drugs herbal origin mentioned in the classical texts in Unani System of Medicine (Table 1). Some of these drugs have beneficial effects in treating the disease in the experimental animals (Table 2) and humans in the clinical situations (Table 3). Both in vivo and in vitro methods have been developed to assay the antiamoebic activity of the herbal drugs (single botanicals as well as multi- component formulations). In in vivo antiamoebic assays rats are used for determination of activity against intestinal infections while hamsters are used for evaluating activity against hepatic infections. E. histolytica is introduced into the caecum via rectum and the. intestine is examined for the presence of amoebae and ulceration. Liver infections are initiated by injection of amoebae into the 10bes.However, in vivo tests are difficult to perform, time- consuming and are unpleasant for the animals. For in vitro antiamoebic assays the development of axenic cultures of Entamoeba histolytica has enabled the development of in vitro assays. Before the development of axenic media, it was only possible to grow the amoebae in presence of bacteria (polyxenic culture) and it made interpretation of the results extremely difficult. Now-a-days, E. histolytica is grown in 96-well microculture plates in the presence of serial dilution of herbal drug extracts or isolated compounds. Atthe end of the test time, the growth of amoebae may be evaluated by visual observation with a microscope. Alternatively,a colorimetric method may be used which involves the removal of culture medium thus, leaving healthy amoebae attached to the bottom of wells while the dead amoebae are washed away. A measure of the number of amoebae remaining in the well is obtained by fixing and staining the parasites. The quantity of stain taken up is proportional to the number of amoebae and can be determined by spectrophotometric method. A relatively simpler method i.e., micro dilution technique for the assessment of in vitro activity against Entamoeba histolytica has been developed and validated with metronidazole. This test has been used to detect the antiamoebic activitiesofextracts of Brucea javanica (Linn.) Merrill fruits and Quassia amara Linn. stems. The activity was found to be associated with quassinoid-containing fractions. The 50% inhibitory concentrations for some quassinoids against amoebae were determined by using this microdilution method. These concentrations ranged from 0.019 j.lglmlfor bruceantin, the most active quassinoid, to greater than 5 j.lgl ml for glaucarubol, the least active compound tested. The micro dilution technique, being more accurate and precise will be quite useful in searching the novel antiamoebic drugs. Natural Product Radiance Vol 2(1) January-February 2003
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Used in
Unan; S,S'•• 01"
I.I- -lul"n
Shakir Jamil*, Shoaib Ahmad**, Jamal Akhtar and Khursheed Alam
Dept. of Moalijat, Facultyof Unani Medicine, lamia Hamdard, New Delhi-l10062, India
There are some 42 million cases annuallyand an estimated 75,000 deaths across the
world due to this disease. In allopathic
systemofmedicine the disease is generallytreated with metronidazole, although this
drug is poorly tolerated by some patients.
There are a number of antiamoebic drugsherbal origin mentioned in the classical
texts in Unani System of Medicine
(Table 1). Some of these drugs havebeneficial effects in treating the disease inthe experimental animals (Table 2) andhumans in the clinical situations
(Table 3). Both in vivo and in vitro
methods have been developed to assay the
antiamoebic activity of the herbal drugs(single botanicals as well as multicomponent formulations).
In in vivo antiamoebic assays
rats are used for determination of activity
against intestinal infectionswhile hamsters
are used for evaluating activity against
hepatic infections. E. histolytica isintroduced into the caecum via rectum
and the. intestine is examined for the
presence of amoebae and ulceration. Liver
infections are initiated by injection ofamoebae into the 10bes.However,in vivo
tests are difficult to perform, timeconsuming and are unpleasant for theanimals.
For in vitro antiamoebic assaysthe development of axenic cultures of
Entamoeba histolytica has enabled the
development of in vitro assays.Before the
development of axenic media, it was only
possible to grow the amoebae in presenceof bacteria (polyxenic culture) and it
made interpretation of the results
extremely difficult. Now-a-days, E.histolytica is grown in 96-well
microculture plates in the presence of
serial dilution of herbal drug extracts orisolated compounds. Atthe end of the test
time, the growth of amoebae may beevaluated by visual observation with a
microscope. Alternatively,a colorimetricmethod may be used which involves the
removal of culture medium thus, leavinghealthy amoebae attached to the bottom
of wells while the dead amoebae are
washed away. A measure of the number
of amoebae remaining in the well is
obtained by fixing and staining the
parasites. The quantity of stain taken up isproportional to the number of amoebae
and can be determined byspectrophotometric method.
A relatively simpler method i.e.,
micro dilution technique for the
assessment of in vitro activity againstEntamoeba histolytica has been
developed and validated withmetronidazole. This test has been used to
detect the antiamoebic activitiesofextracts
of Brucea javanica (Linn.) Merrillfruits and Quassia amara Linn. stems.
The activity was found to be associated
with quassinoid-containing fractions. The
50% inhibitory concentrations for somequassinoids against amoebae were
determined by using this microdilution
method. These concentrations rangedfrom 0.019 j.lglmlfor bruceantin, the most
active quassinoid, to greater than 5 j.lgl
ml for glaucarubol, the least active
compound tested. The micro dilution
technique, being more accurate and
precise will be quite useful in searchingthe novel antiamoebic drugs.
Tinospora cordifolia (Willd.) Miersex Hook. f. & Thoms., Terminalia
chebula Retz. and Zingiber
Antiamoebic compounds fromUnani Drugs
Although a large number of
natural products have been shown to be
able to inhibit the growth of amoebae, very
fewhave been shown to be selectivelytoxic
to the parasite.
Berberine, a benzylisoquinolinealkaloid common in members of the
Menispermaceae, has been clinicallyusedin the treatment of leishmeniasis.
Berberine has been reported to be
effectiveagainst E. histolytica in vitro.The plant flavonoids{ (-) -epicatechin,
(- ) -epigallocatechin and kaempferol}
have been found to higWyactive againstamoebae.
Conessine is one of the steroidal
alkaloids from the bark ofHolarrhena
antidysenterica (Linn.) Wall. and has
also shown an in vitro activityagainst E.histolytica. Emetine from Cephaelisipecacuanha (Brot.) A. Rich.(Rubiaceae) has been found to be higWyactive in the treatment of both hepatic andintestinal amoebiasis but has some toxic
effects especially on the heart. It inhibits
protein synthesis which is probably
responsible for its antiamoebic action andthe toxic effects seen in man. Several
alkaloids from Strychnos spp. have alsobeen found to be active against amoebae
but in contrast to emetine, they were lesstoxic to cells. Usambarensine,
usambarine, and 18,19
dihydrousambarine from Strychnosusambarensis have been found to be
higWy active against E. histolytica invitro. Nb-Methylusambarensine was
comparatively less activeagainst amoebae
than was usambarensine. Gossypol, a
polyphenol from cotton seed oil, is also
Conclusion
rate of efficacy in terms of clinical 4.
improvement.
Casinovi C.G., Fardella G. andGandolini G. (1981). PharmEducation & Science, 36(2),116-22.
Chopra R.N.,Chopra I. C.,Handa KL.
and KapurL.D.,Indigenous Systemof Medicine, U.N. Dhur and Sons
Zaght-al-Dam Qavi for hypertensionAclinical evaluation of the efficacyof the polyherbal Unani formulation, Zaght-al-Dam Qavi in hyperten
sion has been done at lamia Hamdard, Hamdard University,NewDelhi. This Unani formulation consists of Tukhm-e-kahu(Lactuca scariola Linn.), Gul-e-Neelofar (Nymphaea alba Linn. ),Asrol (Rauwolfia serpentina Linn.) andKishneez (Coriandrum sativum Linn.). One dose offormulation containing 109 Kishneez, Tukhm-E-Kahu,lOg,
Gul-e-Neelofar, 5 g and Asrol (root) 19was given orally twice a day for the period of 2 months. The formulation wasfound to be effective in the reduction of clinical symptomps of hypertension in the patients i.e., headache, palpitation,
dizziness, fatigue, nervousness, insomania, etc. The blood pressure was normalized after two months of the treatment