Download Annexure – II

Rajiv Gandhi University of Health Sciences Karnataka,
Bengaluru
“RENOPROTECTIVE ACTIVITY OF AMORPHOPHALLUS PAEONIIFOLIUS
AGAINST GENTAMICIN-INDUCED NEPHROTOXICITY IN RATS”
A Protocol submitted to Rajiv Gandhi University of Health Sciences Karnataka,
Bengaluru
In partial fulfillment of the requirement for the award of
MASTER OF PHARMACY
IN
PHARMACOLOGY
P. MADHURIMA
Department of Pharmacology,
National College of Pharmacy,
Balraj-Urs Road,
Shimoga-577 201
Karnataka-INDIA
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA, BENGALURU
Annexure – II
PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION
Name and Address of the
Candidate
P.MADHURIMA
C/0 P.SATYANARAYANA
PLOT NO. 40, KOUNDINYA NAGAR,
NACHARAM VILLAGE, HYDERABAD,
ANDHRA PRADESH, PIN- 500072.
02
Name of the Institution
National College of Pharmacy,
Balraj-Urs Road,
Shimoga-577 201
Karnataka-INDIA
03
Course of the Study
Branch
M. Pharm. (Pharmacology)
04
Date of Admission to course
04-10-2010
Title of the Topic
“RENOPROTECTIVE ACTIVITY OF
AMORPHOPHALLUS PAEONIIFOLIUS
AGAINST GENTAMICIN-INDUCED
NEPHROTOXICITY IN RATS”
Brief resume of the intended work
6.1. Need for the Study
Enclosure – I
6.2. Review of the Literature
Enclosure – II
6.3. Objective of the Study
Enclosure – III
Materials and Methods
7.1. Source of data
Enclosure – IV
7.2. Methods of collection of data
7.3. Does the study require any
Investigations on animals?
If yes give details
7.4. Has ethical clearance been
obtained form your institution
in case of 7.3.
Enclosure – V
List of References (About 4 – 6)
Enclosure – VII
01
05
06
07
08
Enclosure – VI
Enclosure – VI – A
09
10
Signature of the Candidate
Remarks of the Guide
The present research work is original and not
published in any of the journals. This work
can be carried out in our Pharmacology
Department laboratory.
Name and Designation of
(in Block Letters)
11.1. Guide
Dr. K. L. MANKANI
Ph.D.
Professor and Head,
Department of Pharmacology,
National College of Pharmacy,
Balraj-Urs Road,
Shimoga-577 201
Karnataka-INDIA
11.2.Signature
11.3.Co-Guide (if any)
N. A
11.4.Signature
N. A
11
11.5. Head of the Department
Dr. K. L. MANKANI
Ph.D.
Professor and Head
Department of Pharmacology,
National College of Pharmacy,
Balraj-Urs Road,
Shimoga-577 201
Karnataka-INDIA
11.6. Signature
Remarks of the Principal
12
12.1. Signature
The present study is permitted to perform in
the Pharmacology Department laboratory of
our Institution and the study protocol has
been approved by IAEC.
Principal
Enclosure – I
Brief resume of intended work:
6.1. NEED FOR STUDY:
Nephrotoxicity is caused by several xenobiotic substances damaging
renal proximal tubule, the portion of nephron with greater sensitivity to nephrotoxic
effects. Alcohol is the commonly consumed xenobiotic. Some of the agents which cause
damage to the proximal tubule are anti-bacterial agents such as cephaloridine and
aminoglycosides, anticancer drugs such as cisplatin and industrial chemicals such as
cadmium, hexavalent chromium, mercury and palladium1. Cisplatin decreases
antioxidants and antioxidant enzymes2. Nephrotoxicity is the major side effect of
aminoglycosides, especially gentamicin, accounting for 10 to 15% of all the cases of
acute renal failure1. The defective drug excretion adversely affects the different systems
of the body. Since kidneys are the major organs of drug excretion, occurrence of
nephrotoxicity is of great concern.
Herbal products have a special place in the world of pharmaceuticals. Side
effects of conventional medicines, efficiency of plant-derived drugs and growing interest
in natural products have increased scientific interest in medicinal plants1. Herbal remedies
have been developed through traditional knowledge of herbs, which is a ray of hope for
kidney failure patients3. A number of herbs traditionally used are Kalanchoe pinnata4,
Benincasa cerifera5, Ocimum sanctum6, Phyllanthus niruri7, Passiflora foetida8,
Hemidesmus indicus9 and Ficus racemosa10. Active oxygen species and free radicals play
an important role in several diseases such as rheumatoid arthritis, and cardio-vascular
diseases including cancer11. The antioxidant defense enzymes have been suggestive of
playing an important role in maintaining physiological levels of oxygen and hydrogen
peroxide and eliminating peroxides generated from inadvertent exposure to xenobiotics
and drugs. Any natural compound with antioxidant properties may help in maintaining
health when continuously taken as components of dietary foods, spices or drugs11.
Amorphophallus paeoniifolius or Elephant foot yam is basically a crop of
Southeast Asian origin. It grows in wild form in Philippines, Malaysia, Indonesia and
other Southeast Asian countries. In India it is grown mostly in West Bengal, Kerala,
Karnataka, Andhra Pradesh, Maharashtra and Orissa. In India it is popularly known as
Jimmikand, Suran, Chenna, Ol in Bengali and Oluo in Odia language12.
Amorphophallus paeoniifolius belongs to the family Araceae. A stout cormous
herb, corm depressed globose, 25-30 cm across. Stem is a tuber depressed-globose, darkbrown, root scars prominent, annulate, offlets produced every season, thick and
rhizomatous. Leaf is solitary or two; petiole to ca. 2 meters long and 20 cm in diameter,
background color pale to dark green or blackish green. Lamina is highly dissected , to
about 3 meters in diameter, leaflets rounded, oval, ovate, obovate, elliptic, elliptic-oblong,
elliptic-lanceolate or lanceolate, acuminate, 3-35 cm long, 2 to 12 cm in diameter, upper
surface midgreen, lower surface midgreen to pale green. In flowers inflorescence is shortpeduncled, peduncle is 3 to 20 cm long, about 1 to 8 cm in diameter, usually paler and
smoother than petiole; spathe campanulate, broader than long, 10 to about 40 cm long, 15
to about 60 cm in diameter, base and limb often separated by a shallow constriction, limb
spreading, strongly undulate, base outside very variable, background color ranging from
pale green to dark brown, usually with large and small, circular paler spots, base inside
lower part deep maroon, upper zone dirty whitish or pale pinkish, limb outside as base but
with more prominent maroon flushes, especially near the margin, limb inside usually
glossy dark maroon, base within densely verrucate, verrucae variable, mostly conical,
fleshy. Spadix is sessile, shorter or longer than spathe, 7 to about 70 cm long. Fruits
produced in spikes. Individual fruits are ellipsoid to globular, orange to red.
Seeds are almost as large as fruits. This species requires warm and humid weather
for vegetative growth and cool weather for development of corms. It can be hence grown
as seasonal though it is a perennial. The corms are aperient, carminative and expectorant.
Fresh corms are acrid, stimulant and increase appetite and taste. They are applied
externally as an irritant to treat rheumatism. The corms are administered internally in the
treatment of dysentery, piles and haemorrhoids. Corms are also used in cases of earaches,
swelling of throat, pimples, intercostals neuralgia and fever. It is also used in cases of
elephantiasis, tumors, inflammations, constipations, seminal weakness, fatigue, anemia,
renal disorders and general disability13.
The renoprotective activity of Amorphophallus paeoniifolius tuber has not been
reported so far scientifically. Hence the present study is aimed to investigate
renoprotective activity of extracts of Amorphophallus paeoniifolius.
Enclosure– II
6.2. Review of literature:
1. Therapeutic effect of ethanolic extract of Hygrophila spinosa T. Anders on
gentamicin-induced nephrotoxicity in rats1.
2. Nephroprotective activity of ethanolic extract of dried fruits of Pedalium murex
Linn2.
3. Renoprotective effect of Andrographis paniculata (Burm. f.) Nees in rats3.
4. Protective effect of Kalanchoe pinnata pers. (Crassulaceae) on gentamicininduced nephrotoxicity in rats4.
5. Renoprotective
activity
of
Benincasa
cerifera
fruit
extract
on
ischemia/reperfusion- induced renal damage in rats5.
6. Effect of Ocimum sanctum (OS) aqueous leaf extract on gentamicin induced
nephrotoxicity in rats6.
7. Effect of Phyllanthus niruri Linn. treatment of liver, kidney and testes in CCl4
induced hepatotoxic rats7.
8. In vitro callus induction and in vivo antioxidant activity of Passiflora foetida L.
leaves8.
9. Renoprotective effect of Hemidesmus indicus, a herbal drug used in gentamicininduced renal toxicity9.
10. Modulatory effect of Ficus racemosa: Diminution of Potassium Bromate- Induced
Renal Oxidative Injury and Cell Proliferation Response10.
11. Evaluation of Analgesic activity of methanolic extract of Amorphophallus
paeoniifolius tuber by tail flick and Acetic acid-induced Writhing Response
method14.
12. Pharmacognostic evaluation and Phytochemical Analysis of the tuber of
Amorphophallus paeoniifolius15.
13. Anti-inflammatory
activity
of
Methanolic
extract
of
Amorphophallus
paeoniifolius and its possible mechanism16.
14. Antioxidant potential of Amorphophallus paeoniifolius in relation to their
phenolic content17.
15. Effects of petroleum Ether extract of Amorphophallus paeoniifolius tuber on
Central Nervous System in mice18.
16. Cytotoxic activity of Amorphophallus paeoniifolius tuber extracts In-vitro19.
17. Evaluation of Anthelmintic activity of the Methanolic extract of Amorphophallus
paeoniifolius tuber20.
18. Phytochemical investigation and Chromatographic evaluation of the different
extracts of tuber of Amorphophallus paeoniifolius (Araceae) 21.
19. Anti-diarrheal activity of leaves of Amorphophallus paeoniifolius in Experimental
animals22.
Enclosure – III
6.3. Objectives of study:
1. Collection and authentication of Amorphophallus paeoniifolius plant.
2. Drying of plant material.
3. Extraction of tubers with suitable solvent.
4. Phytochemical investigations of extracts.
5. Screening of renoprotective property by investigation of
a) Serum enzyme estimation and
b) Renal oxidative stress parameters.
Enclosure – IV
MATERIALS AND METHODS:
7.1. Source of data:
The required data will be obtained from:
1. Electronic data [internet]
2. Published research papers
3. Review and research articles from journal
4. Library, National College of Pharmacy, Shimoga, Karnataka, India.
Enclosure – V
7.2. Method of collection of data:
1. The plant Amorphophallus paeoniifolius, will be collected from local areas of
Shimoga district, Karnataka.
2. Screening of pharmacological activity- Renoprotective activity.
Renoprotective activity will be carried out by inducing renal damage with
gentamicin. Histopathological examination will be done. Kidney homogenate will be
prepared and evaluated for renal oxidative stress parameters. Estimation of serum
enzymes, urea and creatinine will be carried out. Activity will be concluded by
comparing the results of control, standard and test samples.
The results will be analysed by ANOVA test.
Enclosure – VI
7.3. Does the study require any investigation or intervention to be conducted on
patients or other humans or animals?
As per the standard procedure Renoprotective activity of the plant Amorphophallus
paeoniifolius will be carried out on wistar rats and albino mice.
7.4. Has ethical clearance been obtained from your institution?
IAEC has approved the present study.
Clearance number: NCP/IAEC/CL/21/05/2011-12.
Enclosure –VII
List of references:
1. Bibu K J, Joy A D and Mercey K A, “Therapeutic effect of ethanolic extract of
Hygrophila spinosa T. Anders on gentamicin-induced nephrotoxicity in rats”, Indian
Journal of Experimental Biology; 2010, 48: 911-917.
2. Shelke T T, Kothai R, Adkar P P, Bhaskar V H, Juvale K C, Kamble B B and Oswal
R J, “Nephroprotective activity of ethanolic extract of dried fruits of Pedalium Murex
Linn.”, Journal of Cell and Tissue Research; 2009, 9(1): 1687-1690.
3. Pratibha Singh, Man Mohan Srivastava and Lakhu Dev Khemani, “Renoprotective
effects of Andrographis paniculata (Burm. f.) Nees in rats”, Upsala Journal of
Medical Sciences; 2009, 114: 136-139.
4. Gaurav Vijay Harlalka, Chadragauda Raosaheb Patil and Mahesh Ramu Patil,
“Protective effect of Kalanchoe pinnata pers. (Crassulaceae) on gentamicin-induced
nephrotoxicity in rats”, Indian Journal of Pharmacology; 2007, 39(4): 201-205
5. Bhalodia Y, Kanzariya N, Patel R, Patel N, Vaghasiya J, Jivani N and Raval H,
“Renoprotective activity of Benincasa cerifera fruit extract on ischemia/reperfusioninduced renal damage in rat”, Iranian Journal of Kidney Diseases; 2009, 3(2): 80-85.
6. Muglikar A G, Kothekar M A, Chilwant K S, Jaju J B and Mateenuddin M D, “Effect
of Ocimum sanctum (OS) aqueous leaf extract on gentamicin induced nephrotoxicity
in rats”, Indian Journal of Pharmacology; 2004, 36(2): 123.
7. Manjrekar A P, Jisha V, Bag P P, Adhikary B, Pai M M, Hegde A and Nandhini M,
“Effect of Phyllanthus niruri Linn. treatment of liver, kidney and testes in CCl4
induced hepatotoxic rats”, Indian Journal of Experimental Biology; 2008, 46: 514520.
8. Rasool S N, Jaheerunnisa S, Jayaveera K N and Suresh Kumar C, “In vitro callus
induction and in vivo antioxidant activity of Passiflora foetida L. leaves”,
International Journal of Applied Research in Natural Products; 2011, 4(1): 1-10.
9. Mangala S Kotnis, Prateek Patel, Sashikumar Narayan Menon and Ramesh Trimbak
Sane, “Renoprotective effect of Hemidesmus indicus, a herbal drug used in
gentamicin-induced renal toxicity”, Nephrology; 2004, 9(3): 142-152.
10. Naghma Khan and Sarwat Sultana, “Modulatory effect of Ficus racemosa:
Diminution of Potassium Bromate- Induced Renal Oxidative Injury and Cell
Proliferation Response”, Basic and Clinical Pharmacology and Toxicology; 2005,
97(5): 282-288.
11. Paliwal R, Sharma V, Pracheta, Sharma S, Yadav S and Sharma S, “Antinephrotoxic effect of administration of DMBA-induced renal carcinogenesis in
Swiss albino mice”, Biology and Medicine; 2011, 3(2): 27-35.
12. http://en.wikipedia.org/wiki/herbalism
13. http://keys.trin.org.au:/key-Server/data/
14. Yadu Nandan Dey, Shankhajit De and Ajoy Kumar Ghosh, “Evaluation of Analgesic
activity of methanolic extract of Amorphophallus paeoniifolius tuber by Tail Flick
and Acetic acid-induced Writhing Response method”, International Journal of Pharma
and Bio Sciences; 2010, 1(4): 662-668.
15. Yadu Nandan Dey and Ajoy Kumar Ghosh, “Pharmacognostic Evaluation and
Phytochemical analysis of the tuber Amorphophallus paeoniifolius”, International
Journal of Pharma. Research and Development; 2010, 2(9): 44-49.
16. Shankhajit De, Yadu Nandan Dey and Ajoy Kumar Ghosh, “Anti-inflammatory
activity of methanolic extract of Amorphophallus paeoniifolius and its possible
mechanism”, International Journal of Pharma and Bio Sciences; 2010, 1(3): 1-8.
17. Jayaraman Angayarkanni, Kunga Mohan Ramkumar, Ulaganathan Priyadarshini and
Poornima Ravendran, “Anti-oxidant potential of Amorphophallus paeoniifolius in
relation to their Phenolic content”, Pharmaceutical Sciences; 2009, 48(6): 659-665.
18. Das S S, Malini Sen, Dey Y N, De S and Ghosh A K, “Effects of Petroleum ether
extract of Amorphophallus paeoniifolius tuber on Central Nervous System in mice”,
Indian Journal of Pharmaceutical Sciences; 2009, 71(6): 651-655.
19. Angayarkanni J, Ramkumar K M, Poornima T and Priyadarshini U, “Cytotoxic
activity of Amorphophllus paeoniifolius tuber extracts In-vitro”, American-Eurasian
Journal of Agriculture and Environmental Sciences; 2007, 2(4): 395-398.
20. Yadu Nandan Dey and Ajoy Kumar Ghosh, “Evaluation of Anthelmintic activity of
the methanolic extract of Amorphophallus paeoniifolius tuber”, International Journal
of Pharmaceutical Sciences; 2010, 1(11): 117-121.
21. De S, Dey Y N and Ghosh A K, “Phytochemical Investigation and Chromatographic
Evaluation of the different extracts of tuber of Amorphophallus paeoniifolius
(Araceae)”, International Journal on Pharmaceutical and Bio Medical Research; 2010,
1(5): 150-157.
22. Purwal L, Shrivastava V and Jain U K, “Studies on Anti-diarrheal activity in leaves of
Amorphophallus paeoniifolius in Experimental animals”, International Journal of
Pharmaceutical Sciences and Research; 2011, 2(2): 468-471.