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Der Pharmacia Sinica, 2011, 2 (1): 129-134

ISSN: 0976-8688
CODEN (USA): PSHIBD

Studies on diuretic activity of three plants from Menispermaceae family

K.K Hullatti1*, M.S Sharada2 and I.J Kuppasth3

1
Department of Pharmacognosy, KLE’s College of Pharmacy, Belgaum, INDIA
2
Department of Studies in Botany, University of Mysore, Mysore, INDIA
3
Department of Pharmacology, National College of Pharmacy, Shimoga, INDIA
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ABSTRACT
This study evaluated the comparative diuretic potential of methanolic root extracts of Cissampelos
pareira, Cyclea peltata and Stephania japonica (Fam: Menispermaceae) in saline primed normal rats
after oral administration. Methanolic root extracts of Cissampelos pareira, Cyclea peltata and Stephania
japonica (100 and 200 mg/kg P.O) or the reference drug, furosemide (20 mg/kg P.O) were administrated
to male Wistar rats (Modified Lipschitz method) and their urine output and electrolyte changes were
quantitated at several intervals of time after the dose for 5 h. All the extracts displayed dose-dependent
diuretic effects. At 100mg/kg both Cyclea peltata and Stephania japonica root extracts shown significant
increase in urinary out put (p < 0.001). However At 200 mg/kg dose all the three extracts significantly
increased (p < 0.001) urine volume and urinary electrolytes, qualitatively similar to that of furosemide.
The methanolic root extract of Cyclea peltata has shown highest diuretic activity (Diuretic action – 3.5).
These findings collectively indicate that the extracts exhibited significant diuretic activity, providing
evidence, at least in part, for their Aurvedic use. Especially root of Cyclea peltata has got highest diuretic
action among three tested extracts. These results support the use of C. peltata roots as “Patha” in
Ayurvedic system of medicine for the treatment of urinary related disorders.

Key words: Cissampelos pareira, Cyclea peltata, Modified Lipschitz method, Patha, Stephania
japonica.
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INTRODUCTION

Diuretics are the drugs used in the treatment of hypertension, congestive cardiac failure, ascites
and pulmonary diseases. The presently available diuretics such as thiazides and furosemide
exhibit various adverse effects such as electrolyte imbalance, metabolic alterations etc. [1]. Some
of oral diuretics are derived from medicinal plants. The Indian Ayurvedic system of medicine is
rich in treating renal problems. Hence a vast number of medicinal plants mentioned in Ayurvedic
system of medicine are known to possess diuretic properties such as Abelmoschus esculentus

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(Bhindaka), Bacopa monnieri (Brahmi), Barbarea vulgaris (Cress), Boerhavia diffusa
(Punarnava), Emblica officinalis (Amla) etc. [2].

The roots of Cissampelos pareira L. Var. hirsuta (DC.), Cyclea peltata (Lam.) J. Hooker and
Thoms., and Stephania japonica (Thunb.) of Menispermaceae family are known as Patha, in
Ayurveda, which is used in the treatment of various diseases like stomach pain, fever, skin
conditions, cardiac pains etc. [3]. The root extract of Cissampelos pareira has been tested for
diuretic activity and its moderate diuretic activity has been reported [4]. The roots of Cyclea
peltata have been tested for their stone formation inhibition activity on ethylene glycol induced
nephrolithiasis in rats [5]. The roots of Stephania japonica are regarded as substitute for
Cissampelos pareira and used in the treatment of fever, diarrhoea and urinary diseases [6]. No
scientific data is available related to diuretic activity or any other urinary related disorders
Stephania japonica.

The various literatures suggests that the Ayurvedic drug Patha is used as diuretic, and the roots
of above said three plants have been used as source of this drug, hence it was decided to evaluate
comparative diuretic activity for these three plant materials.

MATERIALS AND METHODS

Plant material
The roots of Cissampelos pareira Var. hirsuta were collected from dry deciduous forests of
Chamundi hills, Mysore. Cyclea peltata and Stephania japonica roots were collected from
evergreen forests of Madikere. The taxonomical identification of the plants were done by Dr. G.
Shivamurthy, Department of studies in Botany, University of Mysore, Mysore, and the voucher
specimens were deposited at the herbarium (specimen No. KKH-001/2006, KKH-002/2006 and
KKH-003/2006).

Preparation of extracts
The roots were washed with water and shade dried. The dried materials were powdered and
passed through No. 10 sieve. The powdered plant materials were extracted with methanol as
solvent in a soxhlet apparatus. The extracts were concentrated using a rotary flash evaporator
(Superfit, Ambala, India) and semisolid mass was obtained (11.6%, 15.8% and 9.1% w/w with
respect to the powdered material). The extracts were stored in refrigerator until use and the
required quantity was suspended in 1% (w/v) Tween 80 for pharmacological studies.

Experimental animals
The experiment was initiated after approval of Institutional Animal Ethical Committee [(IAEC)
(NCP/IAEC/2/06-07)]. Adult male Wistar rats of either sex weighing 250-300 g procured from
the animal house of National College of Pharmacy, Shimoga, India and were used for the
activity. The animals were maintained under standard conditions as per OECD guidelines
(Temperature 25±2oC) with dark and light cycle (14/10 hrs) and fed with standard dry pellets and
water ad libitum throughout the experiment.

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Standard drug
Furosemide a high ceiling loop diuretic, in the dose of 20 mg/kg BW [7] was used as the
reference drug (Positive control). It was dissolved in water prior to administration.

Biochemical methods
Sodium and potassium levels in the urine were quantitated by flame spectrophotometry
(Systronics flame photometer-129) and chlorides by spectrophotometry (Autochem nexgen
span).

Toxicity studies
Acute toxicity for the determination of LD50 value was performed with different doses of the
extract according to the up and down method using Swiss albino mice of either sex weighing 20
– 25 g. [8].

Diuretic activity
The animals were divided into eight groups of six animals each. Animals were fasted overnight
with water ad libitum and subjected to pharmacological studies. Before treatment, all animals
received physiological saline (0.9% NaCl) at an oral dose of 25 ml/kg body weight (BW). The
first group served as the control and the second group was treated with an oral dose of 20 mg/kg
BW of furosemide. Third and fourth groups were treated with an oral dose of 100 mg/kg and 200
mg/kg BW of methanolic extract of Cissampelos pareira respectively. Similarly, fifth and sixth
groups were treated with methanolic extract of Cyclea peltata while seventh and eighth groups
with methanolic extract of Stephania japonica. All the animals were placed in individual
metabolic cages. Urine was collected and measured at 1, 2, 3, 4 and 5 h after the dose. Sodium
and potassium concentration in urine samples was determined at 5th h by standard biochemical
methods. The ratio of urinary excretion in the test group to that in the control group was used as
a measure of the diuretic action for the given dose of the drug. The diuretic activity was
calculated by comparing diuretic action of extract to that of the standard drug [9].

Statistical analysis
Statistical analysis was analyzed using Student’s t-test for unpaired data. p-Values less than 0.05
were considered statistically significant and less than 0.001 as highly significant. Results are
expressed as mean value ± SEM.

RESULTS

Toxicity studies
All the three extracts did not show any kind of toxic effect when administered up to a dose of
2000mg/kg BW.

Diuretic activity
Effect on urine volume
Results are shown in fig. 1. The methanolic extracts of the roots of Cissampelos pareira at a dose
of 100 mg/kg BW did not show marked diuresis during the 5 h of the test (Cissampelos pareira
3.65±0.19mL versus control 3.33±0.23 mL; P < 0.05).

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Table 1 Effect of methanolic root extracts of C. pareira, C. peltata and S. japonica on urine excretion and ionic
concentration in rats

Cumulative Electrolyte concentration (mmol/l)

Vol. of urine Diuretic
Treatment Dose (ml/kg BW) action Na+ K+ Cl- Na+/K+
Saline 25 ml/kg 3.33±0.23 1.0 85.45±1.13 55.23±0.83 111.50±1.64 1.55±0.04
Furosemide 20mg/kg 14.26±0.78** 4.3 132.68±0.40** 89.83±1.17** 134.50±1.64** 1.48±0.02
C. pareira 100mg/kg 3.65±0.19* 1.1 87.52±0.57 54.50±2.07 124.00±1.41** 1.61±0.06
200mg/kg 8.93±0.29** 2.7 135.62±0.49** 84.50±1.87** 130.50±2.17** 1.60±0.04
C. peltata 100mg/kg 5.47±0.16** 1.6 86.07±0.38 52.50±1.87 134.67±2.66** 1.64±0.05
200mg/kg 11.73±0.47** 3.5 138.15±0.41** 89.93±1.47** 139.17±1.20** 1.50±0.03
S. japonica 100mg/kg 4.17±0.15** 1.3 85.12±0.26 53.34±1.21 140.34±1.37** 1.59±0.04
200mg/kg 9.13±0.40** 2.7 136.45±0.35** 89.50±1.87** 132.50±1.05** 1.50±0.04
Values are mean ± S.E.M., n = 6, * P < 0.05, ** P < 0.001. In parenthesis the cumulative urine volume compared
with control

Fig.1. Time course of diuresis in rats with methanolic root extracts of C. Pareira, C. peltata, S. japonica,
vehicle and Furosemide

The Cyclea peltata and Stephania japonica root extracts significantly increased urinary output to
that of the control (Cyclea peltata 5.47±0.16 mL and Stephania japonica 4.17±0.15 mL versus
control 3.33±0.23 mL; P < 0.001) but the effect was much less than that of furosemide

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(14.26±0.78 mL versus control 3.33±0.23 mL). However, all the extracts of Cissampelos
pareira, Cyclea peltata and Stephania japonica at the dose of 200 mg/kg BW significantly
increased the urinary output. Urine output continued to be stimulated throughout the study
period, such that the cumulative urinary excretion was significantly higher compared to the
control (Cissampelos pareira - 5.47±0.29 mL, Cyclea peltata- 11.73±0.39 mL and Stephania
japonica- 9.13±0.86 mL versus control 3.33±0.23 mL at 5 h after dose; P < 0.01) (Table 1.).

Effect on urinary electrolyte excretion

The effect of single doses of furosemide (20 mg/kg BW) and the methanolic root extracts of
Cissampelos pareira, Cyclea peltata and Stephania japonica (100 and 200 mg/kg BW) on
electrolyte (Na+ and K+) excretion in the 5 h urine in presented in Table 1. All the three plant
root extracts enhanced the excretion of the electrolytes (P < 0.01) which was almost similar to
that of the furosemide. The Na+ / K+ excretion ratio was almost stable (1.48 to 1.64) in all the
groups.
DISCUSSION

The aim of this study was to investigate the comparative diuretic activity of methanolic root
extracts of Cissampelos pareira, Cyclea peltata and Stephania japonica, As these three plants
are used as sources of “Patha” an important Ayurvedic drug which is used in treatment of
urinary related and heart related disorders [10].

Our current study examined the diuretic potential of Cissampelos pareira, Cyclea peltata and
Stephania japonica, root extracts in rats. Oral route was chosen to meet the way used by people
in traditional medicine. The diuretic action induced by the methanolic plant extracts was also
investigated and compared with a standard reference drug, furosemide and control group
(distilled water). The acute treatment of rats by the plant extracts showed a significant diuretic
activity in a dose-dependent manner. The results showed that the highest dose of Cyclea peltata
possesses a strong diuretic activity when given orally at single dose (Fig. 1). The extract of
Cyclea peltata caused a significant increase in urine output from the first hour as observed with
clinically used loop diuretic, while Cissampelos pareira, and Stephania japonica, extracts
showed comparatively moderate diuretic activity. In comparison with these results, a single dose
of furosemide induced a rapid and significant diuresis within 1 h of administration. The duration
of action with furosemide is brief (less than 4 h when oral administration was performed and 3 h
in case of intravenous administration) [11]. All the three plants demonstrated significant increase
in urinary excretion of Na+ and K+ with the dose of 200mg /kg used in our experiments. These
features suggest that the plant extract is acting in a similar way as furosemide, which increases
urinary output and urinary excretion of sodium by inhibiting Na+/K+/Cl- transporter system in the
thick ascending loop of Henley [12].

CONCLUSION

Out of three extracts, methanolic extract of Cyclea peltata at a dose of 200mg/kg BW has shown
the highest diuretic activity. Since all the three plants are known to contain
Bisbenzylisoquinoline type of alkaloids [13] the possible constituents responsible for the diuretic
activity may be these alkaloids, but however the exact mechanism of action and active principle
need to be investigated. This finding supports the Ayurvedic use of these plants as diuretic

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specially roots of Cyclea peltata. Further studies are in progress to evaluate the diuretic activity
of alkaloidal fractions of these plants.

REFERENCES

[1] S. Gupta, L. Neyses, Eur. Heart J., 2005, 26, 644-649.

[2] C.P. Khare, Indian Medicinal Plants, Springer-Verlag Berlin/Heidelberg, 2007.
[3] S.N. Yoganarasimhan, Medicinal plants of India, Part I, Interline publishing Pvt. Ltd., New
Delhi, 2002.
[4] A. Caceres, L.M. Giron, A.M. Martinez, J Ethnopharmacol, 1987, 19, 33-245.
[5] A.J. Christina, P.M. Lakshmi, M. Nagarajan, S. Kurian, Methods Find Exp Clin Pharmacol,
2002, 24, 77-79.
[6] K.R. Kirtikar, and B.D. Basu, Indian medicinal plants, Vol. I., 2nd ed., Periodical experts
Book Agency, New Delhi, 1991.
[7] A., Agunu, E.M., Abdurahman, G.O., Andrew, Z., Muhammed, J Ethnopharmacol, 2005, 96,
471-475.
[8] M.N. Ghosh, Toxicity studies, In: Fundamentals of experimental pharmacology, 2nd ed. SC
and RC book agencies, Calcutta, 1984.
[9] P.K. Mukherjee, Pharmacological screening of Herbal Drugs In: Quality control of Herbal
drugs, Business horizons pharmaceutical Publishers, New Delhi, 1st ed., 2002.
[10] P.V. Sharma, Dravyaguna – Vijnana, Vol. 2., Chaukhambha Bharati Academy, Varanasi,
India, 2005.
[11] D. Brater, Mécanisme de résistance aux diurétiques—conséquences cliniques. In: Grunfeld,
J.P. (Ed.), Actualités néphrologiques. Jean Hamburger. Flammarion Médecine-Sciences, Paris,
1992.
[12] E.K. Jackson, Drugs affecting renal and cardiovascular function. In: J.C. Hardman, A.G.
Gilman, L.E. Limbird, (Eds.), Goodman and Gilman’s The Pharmacological Basis of
Theraputics, 9th ed. Pergmon press, New York, 1996.
[13] K.K. Hullatti, M.S. Sharada, Phcogmag, 2007, 3(11), 173-176.

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