HISTOPATHOLOGICAL STUDIES OF THE EFFECT OF NAGA PARPAM, A ZINC BASED DRUG OF SIDDHA MEDICINE, IN RATS

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Journal of Cell and Tissue Research Vol. 9(2) 1869-1873 (2009)
ISSN: 0974- 0910 (Available online at www.tcrjournals.com) Original Article
HISTOPATHOLOGICAL STUDIES OF THE EFFECT OF NAGA
PARPAM, A ZINC BASED DRUG OF SIDDHA MEDICINE, IN RATS
ILANGO, B., DAWOOD SHARIEF, S., VINOTH KUMAR,K., RAJKUMAR, R.,
PRATHIBA, D.1 AND SUKUMAR,E.2
Received:May 17, 2009;Accepted: July 4, 2009
School of Environmental Sciences, Post Graduate and Research Department of Zoology, The New College, Chennai
– 600 014, 1Department of Pathology, Sri RamachandraMedical College and Research Institute, Sri
Ramachandra University, Porur, Chennai-600116, 2Department ofApplied Sciences, Higher College of
Technology, P.B.74,P.C.133,Muscat, Sultanate ofOman. E. mail: sdawoodsharief@yahoo.co.in
Abstract: Siddha medicine, one of the oldest traditional medical systems of India, uses metals
and minerals as drugs extensively in addition to plants and animal products to treat several
diseases. Naga Parpam (NP), a metallic preparation containing zinc as the major constituent
and used in treatment of piles, tuberculosis and dysentery, has been chosen for assessing its effect
on the tissues of vital organs in rats. The drug was administrated orally in doses of 5, 10, 20 and
40 mg/kg body weight to the animals for 15, 30 and 60 consecutive days. The results revealed
that in 15 day treatment, liver and kidney had normal histology with no marked changes in the
tissue architecture. In 30 day study, the kidney tissues remained normal while the liver tissues
exhibited a few apoptotic cells with mild focal and lobular inflammation. On 60 day treatment,
liver showed mild lobular inflammation and apoptotic cells were found in all doses. However
brain, kidney and testis remained normal even in higher doses. The results suggest that NP does
not show any toxicity in short term administration which is advocated in Siddha literature for all
metal based drugs.
Key words: Naga Parpam, Siddha medicine, Histopathology
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INTRODUCTION
In India, the traditional medicines viz. Ayurveda,
Siddha and Unani are serving the people to a
considerable extent especially in rural areas and
places not served by allopathic medicine. In Siddha
medicine, a popular system practiced in South India,
metals such as copper, zinc, mercury, silver, gold etc.
are extensively employed in drug preparations that
are used to treat some baffling diseases [1]. These
metals, which exert toxic effects on living organisms
even in small doses, are purified using specialized
traditional techniques by repeatedly triturating with
herbal juices and ashing them in sealed earthenware
containers, to free them from toxicity. Further these
techniques also change their properties and render
them useful as therapeutically active products [2-4].
The present investigation has been undertaken to
examine the toxic effects of Naga parpam on liver
and kidney tissues in 15, 30 and 60 days treatment in
rat model.
MATERIALS AND METHODS
Drug: Naga parpam prepared according to the
procedure mentioned in the traditional Siddha
literature [5] was procured from the Indian Medical
Practitioners’ Co-operative Pharmacy and Stores
(IMPCOPS), Chennai. Analysis of Naga parpam
by S4 pioneer wavelength dispersion X-ray
fluorescence spectrometer (WDXRFS, Bruker-AXS,
Germany) at SophisticatedAnalytical Instrumentation
Facility, IIT, Madras revealed 65.59% of zinc and
2.27% of iron.
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J. Cell Tissue Research
a1 a 2
a 3 a 4
Animals: Male albino rats of Wistar strain (180 –
200 g) obtained from the Tamil Nadu University of
Veterinary and Animal Sciences, Chennai and
maintained according to the guidelines of CPCSEA,
under the supervision of Animal Ethics Committee
were used for the experiment. They were acclimatized
to laboratory conditions prior to use and fed
with pelletted chow (supplied by Poultry Research
Station, Chennai) and water provided ad libitum.
The animals were divided into five groups each
containing 6 rats. Group I served as control and to
the other four groups, the drug NP was orally
administered in doses of 5, 10, 20 and 40 mg/kg body
weight, as a suspension in 1% carboxymethyl
cellulose by intubation for 15 consecutive days. To
find the effect of chronic administration on various
tissues, the drug was also administered to two other
sets of animals in the same manner respectively for
30 and 60 days. The doses for the studies were
chosen based on the preliminary toxicity screening
[6]. The animals were sacrificed by decapitation on
the 16th, 31st and 61st day and the vital organs such
as brain, liver, kidney and testis were removed in
control and drug treated animals, washed with 1%
ice cold saline and fixed in neutral buffered 10%
formalin. The tissues were trimmed and processed
as per the procedure of Pearse [7]. 3-5 μm paraffin
sections were stained with haematoxylin, eosin and
mounted. The slides were examined under light
microscope.
RESULTS
Liver and kidney were selected for evaluation as
they are sensitive to metal toxicity. In the 15 days
treatment set, the tissues of liver and kidney revealed
normal histology with all the doses of NP, and no
changes in the tissue architecture were observed. In
30 days set, the kidney tissues remained normal in all
the doses while liver tissues showed a few apoptotic
cells withmild focal, lobular inflammation in 5mg/kg
dose. With other doses therewas mild periportal inflammation,
lobular inflammation and fewapoptotic cells
(Figs. 1,2).
In 60 days study, NP showed marked changes in the
liver tissues (Fig 3). As the dosage increased from
5-40 mg/kg, mild periportal inflammation, lobular
inflammation, regeneration of hepatocytes and
apoptotic cells were found. However the tissues of
brain, kidney, testis remained normal (Figs. 4-6].
DISCUSSION
Zinc is an essential element and nontoxic in minute
doses [8]. It plays an essential role in cell membrane
integrity and is a component ofmore than 300 different
enzymes of cellular metabolism, involving proteins,
lipids and carbohydrates [9]. The metal has been
reported to interact with cell membranes to stabilize
them against various damaging effects, including
oxidative injuries [10].
Fig. a1: T. S. of liver demonstrating normal
lobules in control rat.
Fig. a2: T. S. of kidney demonstrating
normal structure in control rat.
Fig. a3: T. S. of brain demonstrating normal
structure in control rat.
Fig. a4: T. S. of testis demonstrating
normal structure in control rat.
Abbreviations used in figures: AC – Apoptotic cells, BC – Binucleated cells, CV – Central vein, HC – Hepatic cords, LI –
Lobular inflammation, PI – Periportal inflammation, SN – Sinusoids, CT – Collecting Tubules, BC – Bowman’s Capsule, G –
Glomeruli, CT – Collecting Tubules, DE – Ductus Epididymis, GM – Grey Matter, GL – Granular Layer, PC – Purkinje Cells,
WM –White Matter, AC – Apoptotic Cells, HC – Hepatic Cells, RH – Regenerating Hepatocyte, LI – Lobular Inflammation
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Fig.1 Fig. 2
Fig. 1(f-i): T.S. of liver of rat treated with Naga parpam tract
for 15 days. x 100 (Figs.1 b-e) and 30 days. x 200
b & f - Naga parpam treated with 10mg/kg body weight;
c & g - Naga parpam treated with 20mg/kg body weight;
d & h - Naga parpam treated with 30mg/kg body weight;
e & i - Naga parpam treated with 40mg/kg body weight.
Fig. 2(f-i): T.S. of kidney of rat treated with Naga parpam
tract for 15 days. x 100 (Figs. b-e) and 30 days. x 200
b & f - Naga parpam treated with 10mg/kg body weight;
c & g - Naga parpam treated with 20mg/kg body weight;
d & h - Naga parpam treated with 30mg/kg body weight;
e & i - Naga parpam treated with 40mg/kg body weight
Ilango et al.
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J. Cell Tissue Research
Fig.3 Fig.4 Fig.5 Fig.6
Histology of rat liver (Fig. 3), kidney (Fig. 4), brain (Fig. 5) and testis (Fig. 6) treated with Naga Parpam for 60 days
b - Naga parpam treated with 10mg/kg body weight
c - Naga parpam treated with 20mg/kg body weight;
d - Naga parpam treated with 30mg/kg body weight;
e - Naga parpam treated with 40mg/kg body weight.
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In the present investigation NP in short term
administration did not affect the liver and kidney
tissues as they showed normal histology with no
obvious changes in their tissue architecture. However,
in the 30 days treatment, the liver tissues of animals
treated with 5 mg/kg dose of NP exhibited a few
apoptotic cells with mild focal and lobular inflammation.
In higher doses (10-40 mg/kg), there was
mild periportal and lobular inflammation. These
observations indicated that continuous administration
of NP in high doses may affect the functioning of
vital organs such as liver and kidney.Allen et al. [11]
reported the hepatic lesion formation in tissues of
zinc sulphate treated sheep for 13 days. Fatty
infiltration of liver and nephrosis were reported in 21
day treatment with zinc by Straube et al. [12].
In long termstudies with NP, toxic symptoms of mild
periportal inflammation, lobular inflammation, regeneration
of hepatocytes and apoptotic cells were
observed in the tissues of liver while in rest of organs
there were no appreciable changes. In the earlier
studies by Allen et al. [11] hepatic lesions were
observed in sheep treated with zinc for 72 days.
Similarly histopathological lesions accompanied by
increased liver, kidney and brain weights were
observed in rats on 13 week treatment [13].
The present observations also reinforce the importance
of strict adherence to the prescriptions of
traditional medical practitioners in dosage as well as
duration while using these metallic preparations.Any
over usage or self medication may invariably lead to
adverse reactions as mentioned in the age old
literature.
ACKNOWLEDGEMENTS
The authors thank the Head, Post Graduate and
Research Department of Zoology, Principal and the
Management of The New College, Chennai, India
for providing the necessary facilities and encouragement.
REFERENCES
[1] Subbarayappa, B.V.: Lancet. 350, 1841-1844 (1997).
[2] Said, M. In: Hamdard Pharmacopoeia of Eastern
Medicine. Hamdard Foundation, Karachi, Pakistan,
pp. 233-234 (1969).
[3] Chopra, R.N., Chopra, I.C., Handa., K L., and Kapur,
L.D.. In: Chopra’s Indigenous Drugs of India.
(Chopra, R.N. ed) Academic Publishers, Calcutta,
India, pp 461- 464 (1982).
[4]Vohara, S.B., Kim, H.S., Ganotra, K.B.,Mishra, G.V.,
Ayesha, Bajaj, S. and Athar, M.: Investigations on
the use of Arsenic, Lead, Mercury, Gold and Silver
in Indian Medicines. Abstract. 2nd National
Conference and Workshop of Ethanopharmacology
on 2nd- 4th October at JSS College of Pharmacy,
Mysore, India. (1997).
[5] Anonymous, Formulary of Siddha Medicines, The
IndianMedical Practitioners Co-operative Pharmacy
and Stores Ltd. (IMCOPS),Madras. 12-13: 26-55; 155-
172 (1972).
[6] Ilango, B., Biochemical and Toxicological Evaluation
of Some Metallic Drugs in Siddha Medicine. PhD
Thesis, University ofMadras, India, pp. 57-58 (2007).
[7] Pearse, A.G. Histochemistry: Theoretical and applied,
Vol 1 and 2, (4th edn.), Churchill Livingstone,
Edinburgh (1985).
[8] Bertholf, R.L.. Zinc. In: Handbook on Toxicity of
Inorganic Compounds (Sigel, H. and Seiler, H.G. eds),
Marcel Dekker, Inc., NewYork. pp. 787-800 (1988).
[9] Parkin, G.: Science 305, 1117-1118 (2004).
[10]Bettger,W.J. andO’Dell, B.L.: Life Sci., 28: 1425-1438
(1981).
[11]Allen, J.G.,Master,H.G. and Peet,R.L.: J.Comp.Pathol.
93:363-377 (1983).
[12] Straube,E.F.; Schuster,N.H. andSinclair,A.J.: J.Comp.
Pathol. 90: 355-361 (1980).
[13] Bai, K.M., Krishnakumari,M.K. and Ramesh, H.P.:
Indian J. Exptl.Biol. 18: 854-857 (1980
Ilango et al.