SP-31

NEPHROPROTECTOR ACTIVITY OF ETHANOLIC EXTRACT OF PODS

OFCANAVALIAGLADIATA

ANNAPUREDDYAISHWARYA, T. V E N K A T A S I V A REDDY, GUMMADI ASHA,

UPPULURISPANDANA*

Department of Pharmaceutical Chemistry, Nirmala College of Pharmacy, Atmakur, Mangalagiri, Guntur.

E Mail id: spuppuluri@gmail.com

ABSTRACT

Nephrotoxicity is a major limiting factor in cisplatin treatment. In the present study, hydro-ethanolic pod extract
of canavliagladiatawas investigated for its protective role in cisplatin induced nephrotoxicity. The experiment
was designed for 15 days and healthy adult albino rats were divided into 5 groups. Group 1 received 1% carboxy
methyl cellulose(CMC), in distilled water for 15 days, group 2 received cisplatin (6mg/kg b.w) on 5th day, group
3 received the low dose of C.gladiata (200mg/kg b. w) suspended in the vehicle for 15 days, group 4 received
the high dose of C.gladiata (400mg/kg b. w) suspended in the vehicle for 15 days, group 5 received only
C.gladiata suspended in the vehicle for 15 days, and animals belongs to group 3 and 4 were received cisplatin
(6mg/kg b. w)on the day 5. At the end of the experiment urine samples and blood samples were collected from
all the groups and were sacrificed to study renal functional parameters. Treatment with the C.gladiatapod
extract significantly (0.05) attenuates renal damage by decreasing serum creatinine and blood urea
nitrogen(BUN), enhanced the activities of catalase,GSH, LPO,UTP, CLcr, levels compared with cisplatin
treatment group. Our results suggest that, pod extract of C.gladiata may ameliorate renal damage caused by
cisplatin.

KEYWORDS: Nephrotoxicity, cisplatin implication, Canavaliagladiata, anti-oxidant parameters.

Abbreviations: CAT-catalase, Clcr-creatinine clearance, GSH- Glomerular filteration rate,

LPO-lipid peroxidation, UTP- urinary total protein.

INTRODUCTION

NEPHROTOXICITY
Nephrotoxicity is the toxicity in the kidneys. It is a poisonous effect of some substance, both toxic chemicals and
medications, on renal function. Nephrotoxicity is an intrinisicadverse effect of certain anti cancer drugs.
Anticancer drugs have therapeutic effect but it may also produce nephrotoxicity. 1,2Multiple risk factors exist in
majority of patients with acute renal failure.glomerular filteration rate declined beyond the age of 50, may be
because of vascular changes.3,4

CISPLATIN
Cisplatin is used in the treatment of several solid tumours unfortunally it is also one of the most toxic anti- cancer
drug its dose limiting toxicity being nephrotoxicity.6 Cisplatin produces an acute, mainly proximal tubular
functional impairment within hours administration.7,8

CISPLATIN NEPHROTOXICITY
The most serious and usually dose limiting and usually dose limiting toxicity of cisplatin renal experimental
cisplatin induced nephrotoxicity was first reported in 1971. Nephrotoxicity of cisplatin is associated with its
metabolism.9,10 Nephrotoxic effects of cisplatin has been well documented in all species studied including
mice, rats,dogs,and humans. Acute tubular necrosis is a prominent feature of cisplatin nephrotoxicity and is
clinically manifested by elevation of blood urea nitrogen (BUN) and serum creatinine (SC), proteinuria and
hyperuricemia. 11

Int J Life Sci Pharma Res. ISSN 225-0480; SP-10; “Exploring and Advancing Healthcare through Novel Strategies in Pharmacy Practice” 2019.
Int J Life Sci Pharma Res. ISSN 225-0480; SP-10; “Exploring and Advancing Healthcare through Novel Strategies in Pharmacy Practice” 2019.

CANAVLIA GLADIATA
Canavalia gladiata is a vigorous, deep-rooted, and annual to perennial climbing plant. The stems, which are
often slightly woody, can grow up to10 meters long, scrambling over the ground or twining into other plants
for support .12,13,14

ACTIVITIES OF CANAVALIA GLIADIATE:

 ANTI MICROBIAL ACTIVITY
 HEPATO PROTECTOR ACTIVITY
 ANTI HELMENTIC ACTIVITY
 ANTI BACTERIAL ACTIVITY

MATERIALS AND METHODS

Preparation of extract
The hanging aerial pods were washed thoroughly with distilled water and shade-dried. Ethanolic extract of
the dried roots of canavilia gladiate was prepared by soxheltmethod using. 15,16,17 The ethanoilc extract thus
obtained was dried under reduced pressure at aroom temperature not exceeding 40 oC to get a yield of 90%
from the crude extract. (18,19)

CHEMICALS
All the chemicals used were of analytical grade, from SD, merk [INDIA] chemicals .20,21

ANIMALS
Healthy adult male albino rats [100-150mgbw] of wistar strain aged 60-90 days were used for the study. The
rats were housed in polypropylene cages maintained at standard conditions. The animals had free access to
standard lab chew and tap water. 22

TREATMENT PROTOCOL
Animal were divided in to5 groups each group consisted of six rats [n=6]
 The animals of group-1 recived 1% carboxy methyl cellulose [CMC] in distilled water for 10 days.
 the animals of group-2 recived cisplatin[6mg/kgb.w]on 5th day
 The animals of group-3 recived the low dose of canavliagladiata [200mg/kgb.w] suspended in the
vehicle for 10 days.
 The animal of group-5 recived only canavliagladiata suspended in the vehicle for 10 days.
 Animals which belong to group-3 and 4 were received cisplatin [6mg/kg b.w, i.p] on day 5. Group -1
recived normal saline instead of cisplatin.23,24

BIO CHEMICAL DETERMINATION

On the day9, urine was collected with the help of metabolic cages and the urine samples were subjected for
estimation of urinary functional parameters. The animals were sacrificed by cervical decapitation and blood
samples werecollected by cardic puncture and were used for estimation.25,26

RESULTS
Table - 1
ESTIMATION OF SERUM PARAMETERS
S.NO GROUP BUN SC
1. Control 22.08±0.80 0.6±0.05
2. Ccl4 42.05±1.10 2.25±0.20
3. 200mg of extract 27.01±0.10 1.90±0.10
4. 400mg of extract 29.05±0.05 1.14±0.05
Int J Life Sci Pharma Res. ISSN 225-0480; SP-10; “Exploring and Advancing Healthcare through Novel Strategies in Pharmacy Practice” 2019.

Table - 2
ESTIMATION OF URINARY PARAMETERS:
S.NO GROUP UTP CLcr
1 Control 07.6±0.15 18.80±1.6
2. Ccl4 20.1±0.12 0.510±1.5
3. 200mg of extract 08.4±0.01 13.05±1.2
4. 400mg of extract 0.64±0.15 11.00±0.2

Table-3
ESTIMATION OF TISSUE HOMOGINATION PARAMETERS:
S.NO GROUP GSH LOP CAT
1. Control 145.12±0.1 11.12±0.1 01.25±0.12
2. Ccl4 72.15±2.3 15.12±0.1 08.10±0.57
3. 200mg of extract 85.42±2.7 12.12±0.5 12.05±1.3
4. 400mg of extract 90.10±1.90 12.0±0.0 14.0±1.00
Each value represents mean±SEM from six animals in each group.
P<0.05 is significant when compared with normal group.

DISCUSSION

Cisplatin is one of the most important anti-tumour agents and is highly effective against a diverse spectrum
of malignancies. However, the use of this agent in combating cancer is limited by the development of
nephrotoxicity along with various untoward side effects including nausea, vomiting, diarrhea and
myelosuppression.33,34 The present study was focused on the effects of ethanolic extracts of pods of Canavalia
gladiate on the renal damage induced by cisplatin. The precise mechanism of cisplatin induced nephrotoxicity
has not been elucidated. The basic effect of cisplatin induced toxicity on the cellular membrane is believed to
be peroxidation of membrane lipids. Cisplatin caused renal failure characterised by increase of serum marker
levels, decline in the renal functional parmeters indicated by increased the urinary protein extraction and
decreased the Clcr. The nephroprotector activity in rats was further more concluded by viewing the results of
the following parameters.

1. SERUMMARKERS
BUN: In the cisplatin induced rats the BUN levels are vigorously increased and when the extracts of C. gladiata
were administered the level are lowered and has attained the normal level in case of (400 mg/kg) dose.
SC: In the cisplatin induced rats the SC levels are vigorously increased and when the extracts of C.gladiatawas
administrated the levels are lowered significantly.

2.URINARY FUNCTIONAL PARAMETERS

UTP: In the cisplatin induced rats the UTP levels are vigorously increased and when the extracts of
C.gladiata were administered the levels are significantly lowered.
CLcr: In the cisplatin induced rats are the CLcr levels are lowered down and when the extracts of C.gladiata
were administered the levels are significantly increased.

3. TISSUE HOMOGENENATE PARAMETERS/ESTIMATION OF ANTIOXIDANT ENZYME

ACTIVITY
LPO: In the Cisplatin induced rats the LPO levels are increased than the normal level and when the extracts
of C .gladiata were administered the levels are significantly decreased.
GSH: In cisplatin induced rats the GSH levels are decreased than the normal level and when the extracts of
C.gladiata were administered the levels are significantly increased.
CAT: In the cisplatin induced rats the CAT levels are decreased than the normal level and when the extracts
of C.gladiata were administered the levels are significantly increased.
Hence by viewing the above results we can conclude that, C.gladiata attenuates thenephrotoxicity of
cisplatin in rats and confirm the antioxidant potential of canavaliagladiata.

Int J Life Sci Pharma Res. ISSN 225-0480; SP-10; “Exploring and Advancing Healthcare through Novel Strategies in Pharmacy Practice” 2019.
Int J Life Sci Pharma Res. ISSN 225-0480; SP-10; “Exploring and Advancing Healthcare through Novel Strategies in Pharmacy Practice” 2019.

Graphical Representation

CONCLUSION

When the high doses (400mg/kg) were administered to the rats, no toxicity was observed. So the ethanolic extract
ofpods of C. gladiata have no side effects and also it has the protective effect against cisplatin induced
nephrotoxicity.

ACKNOWLEDEMENTS

The authors are grateful thank to the Management and Principal for providing facilities to do this work.

REFRENCES

1. Abhimananyu Sharma, Asmitha Gupta, Sakshi Singh Amla Batra; Tiosporacordifolia (wild)
Hook.F.&Thomson- A plant with immense economic potential; J.Chem.Pharm. Res., 2010, 2(5); 327-333.
2. Amer MH, Izbicki R, and AI-Sarff M. 1978 T hermozolamide and whole brain irridation in melanoma
metastacic to the brain: a phase 2trial of the cytokine Melanoma and Central Nervous System Metastates.
PROC Am soc Clin.Oncol. 19:312.
3. Anderson ME., Naganum A and Meister, I Balachandran, PN Ravindran; In Vitro Cell. Dev. Biol.-plant, 2006,
42,584-588.
4. Bengtsson, U. (1962). A comparative study of chronic non- obstructive Pyelonephritis and renal papillary
necrosis. Act med. Scand., 388 (supple):1-71.
5. Blum,R.H. and Carter, S.K. (1974): Adriamycin: a new anticancer drug with significant clinical activity. Ann.
Intern Med. 80: 249-259.
6. Bompart G (1989) Cisplatin induced changes in cytochrome P-450, lipid peroxidation and Drug-metabolizing
enzyme activities in rat kidney cortex; Toxial Lett. Aug; 48 (2):193-9.
7. BT Kavitha, SD Shruthi, S Padmalatha Rai, YL Ramachandran; Phytochemical analysis and hepatoprotective
properties of TINOSPORA Cordifolia against carbon tetra-chloride-induced hepatic damage in rats Journal
of basic and clinical Pharmacy;2011.
8. Bunn PA Jr; 1989; the expanding role of cisplantin in treatment of non- small- cell lung cancer; SeminOnocol.
16(4 suppl6); 10-21.
 9. Burch PA; 1999; phase 2 trial of combination paclitaxel and cisplantin in advanced Urothelial Carcinoma (U
C) (Meeting abstract);ASCO AnnualMetting Ab.No.1266.
10. BV Shetty; V Singh; Flora of Rajasthan. 1 st edition, Merrut Publishers and Distributors, Merrut. Vol1.2010;
756-100.
11. Choie DD., LonecherDs. And Dal campo AA. (1981); Acute and chronic cisplantinum nephrotoxicity in rats;
Lab. Invest; 44,397-402.
12. Cok G.Goksel T., sover s., atil h., aysan.,(2006); comparing cisplatin plus etoposide with combination of
mitomycin, ifosfomide and cisplatin in advance non-small cell lung cancer patients;tuberktoraks 54(2):161-
7.
13. Crino l, clerici m, figol,i f , carlini p , ceci g, carpi a,santinia,dicostanzo f, boni c, et al, 1995;chemLotherapy
of advanced non-small-cell lung cancer :a comparision of three active regimens; a ranadmized trail of the
Italian oncology group for clinical research(G.O.I.R.C);ann oncol;6(4):347-53.
14. Cvitkovic,E.SpuauldingJ.,Bethune,V.,Matrin,j.,&Whitmore,W.F.[1997] Improvement of cis dichlorodiaimino
platinum [NSS-119875]Therapeutic index in an animal model.cancer,39;1375-1361. Daugaard G.et al.,
[1988b]: Effects of cisplatin on proximal convoluted and straight segments of the rat kidney. J. Pharmacol.
Exp. Ther., 244:1081-5.
15. Dentino M,LuftFC,YumMW, WilliamsSDandEinhorn LH [1978]; Long term effect of cis-
diamminedichloride Platinum [CDDP] on renal function and structure in man; Cancer;41:1274-81
16. Dobyan Dennis C, Joseph Levi, Charlotte Jacobs, Jon Kosek and Michale W. Winner [1980]; Mechanism of
ciplatinum nephrotoxicity: 2 Morphological observations; J. Phamacol. Exp. Ther.; 213[3]:551-56. D
Sarma, P Padma, RL Khosa; Constituents of Tinosporacordifolia root .Fitoterapis.1998; 69:541-542.
17.D Sarma, RL Khosa, M Sahai; Isolation of Jatrorrhizine from Tinosporacordifoliaroots.Planta Med .1995;
61:98-99.
18. Espana P ,Smith F ,Ahrams J. Haidak D, Veno W, Wooley P ,and Schein P,1984, Phase-2 study of cis-
diamine – dichloro platinum [cisplatinum], Bleomycin Methotrexate for advanced squamous cell
carcinoma of head and neck , cancer chemotheraphy and pharmacology, Vol. 12,2,98-100.
19. Goldstein RS, Noor weir B, Bond JT, Hook JB and Mayor GH [1981]; Cis- D ichloroDiammine- Platinum
nephrotoxicity; time course and dose response of renal functional parameters; Toxicol . Appl . Pharmacol;
15; 60 (2):163-75.
20. Hanneman J and Baumann K (1990). Nephrotoxity of cisplatin, carboplatin and transplatin; A
comparative in vitro study. Arch toxicology. 64(5)393-400.
21. Hanneman J and baumann K (1998).cisplatin induced lipid peroxidation and decrease of gluconeogenesis in
rat kidney cortex: different effects f antioxidants and radical scavengers . Toxicology 51 (2-3). 119-132.
22. Hara,M; Yoshida,M; Nishjima, M ; Ligo , M; Ohatani-Kaneko, r; Shimada, A; Hasegawa, T ; Akama, Y. and
Hirata, K.(2001). Melatonin a pineal secretory product with antioxidant properties, protects against
cisplatin-induced nephrotoxicity in rats, J. pineal.Res. 30:129-138.
23. Higby`D J, Wallace H J Jr,Alburt D, Holland JF. 1974. Diamino dichloro- platinum in the chemothrerapy of
testicular tumors. J Urol; 112; 100-4.
24. JG Asthana, S Jain , A Mishra , MS Vijayakant; Evaluation of antileprotic herbal drug combination with
dapsone Indian Drugs. 2001; 38:82-6
25.J Singh, K Sinha , A Sharma, NP Mishra, SP Khanuja; Traditional uses of Tinosporacordifolia (Guduchi) J
Med A romat plant Sci. 2003;25:748-51.
26.K Avnish Upadhyay, Kaushal kumar, Arvind kumar, and S Hari Mishra. International journal of Ayurveda
Research; 2010 Apr- Jun; 1(2): 112-121.
27. Karen Kelly, John Crowley, Paul a; Bunn Jr; Cary A. Preasant, Patrak, Grevastad, Carol M; MoinPour, ScottD
.ramsay, Antoinette J; Wozniak, Greoffery R; Weiss Dennis F; Noore, Valerie K ; Israel, Robert B
; Livingston and Dravid R. Chandra (2001). Randamised PHASE3 trail of paclitaxel plus Carboplatin
Versus Vinorelbine plus cisplatin in the treatment of patients with advanced non- small- all lung cancer : A
South West Oncology group Trail; J .Clinical Oncology; 19,13; 3210-18.
28. KC Gupta, R Vishwanathan; Antituberculous substances from plants. Antibiotic Chemother.1956; 6:194 -5.
Kleinknecht TH; Mitrou PS; K ochenDorfer KJ; and Bergmann L; and Mery, J.P.(1978) immunologically
mediated drug- induced acute renal failure contrib .Nephron; 10:42-52.
29. KN Aiyer, M kolammal; pharmacognosy of ayuervadic drugs, series1. 1 st ed. Trivendram: the central research
institute; 1963.

Int J Life Sci Pharma Res. ISSN 225-0480; SP-10; “Exploring and Advancing Healthcare through Novel Strategies in Pharmacy Practice” 2019.