Diuretic activity of whole plant extract of Leptadenia reticulata

 

S. Mohanraj1*, Santhoshkumar C. 2 and A. Chandran3

1Jogaiah Institute of Technology and Sciences, College of Pharmacy, Kalagampudi, Andhra Pradesh, India

2Srikrupa Institute of Pharmaceutical Sciences, Siddipet. Andhra Pradesh, India

3PGP College of Pharmaceutical Science & Research Institute, Namakkal, India

 

ABSTRACT:

The present study was undertaken to investigate the diuretic effect of aqueous and ethanolic extract of the Whole plant of Leptadenia reticulata in normal rats. Aqueous and ethanolic extracts of whole plants of Leptadenia reticulata were administered to experimental rats orally at doses of 100mg/kg p.o. Furosemide (100 mg/kg) was used as positive control in study. The diuretic effect of the extract was evaluated by measuring urine volume, sodium, potassium and chloride content. Urine volume was significantly increased by both aqueous and ethanolic extract in comparison to the control group but this effect was less than furosemide. Significant increase in renal clearance of sodium, potassium and chloride ions was observed in treated and standard groups.

 

KEYWORDS: Diuretic activity, Furosemide, Leptadenia reticulata, aqueous and ethanol

 

INTRODUCTION:

Herbal medicine represents one of the most important fields of traditional medicine all over the world. To promote the proper use of herbal medicine and to determine their potential as sources for new drugs, it is essential to study medicinal plants, which have folklore reputation in a more intensified way1. Diuretics are drugs that increase the rate of urine flow, sodium excretion and are used to adjust the volume and composition of body fluids in a variety of clinical situations. Drug-induced diuresis is beneficial in many life threatening disease conditions such as congestive heart failure, nephritic syndrome, cirrhosis, renal failure, hypertension, and pregnancy toxaemia2. Naturally occurring diuretics include caffeine in coffee, tea, and cola, which inhibit Na+ reabsorption and alcohol in beer, wine and mixed drinks, which inhibit secretion of ADH3. Leptadenia reticulata is a much branched twining shrub of family Asclepiadaceae. It is commonly known as Dodi in Hindi, Jivanti in Sanskrit and Palaikodi in Tamil. Flowers are greenish yellow, in many flowered cymes (in lateral or subaxillary cymes), the follicules are sub woody and turgid. Stem is cylindrical and bent occasionally at places. It is 5 to 10 cm long, 0.5 to 2.5cm in diameter. The surface is rough, longitudinally ridged, Wrinkled and forrowed , transversely cracked and with vertically elongated lenticels at places. Externally whitish brown, internally pale brown, fracture short and splintery, odor and taste are not characteristics.The bark is yellowish brown , corky , deeply cracked.Leaves are ovate to cordate ,4 to 7.5 cm long , 2 to 5 cm broad , entire , acute , subacute to mucronate , base symmetrical  , petiole 1 to 3 cm , glabrous above and pubescent below, green color taste and odor not characteristics. The roots are externally rough, white or buff colored with longitudinal ridges.The root size varies from 3 to 10 cm in length and 1.5 to 5 cm in diameter4,5,6,7.


 

 

 

It is widely used in traditional system of medicinand also well known for its vasodilator8, anti-depressant9, anti-abortificant10, Prostatic hyperplasia11, anti-anaphylactic effect12, anticancerous activity13, anti implantation activity14, anti-microbial activity15.

 

MATERIALS AND METHODS:

Plant collection and authentification

The fresh whole plant of Leptadenia reticulata Linn (Asclepiadaceae) for the research work was collected from the roadsides of Narasapur in the month of November 2011 and was authenticated by botanical survey of India Coimbatore and the specimen voucher samples are kept in BSI herbarium for reference BSI/SRC/12/20/2011.

 

Preparation of extracts

500gms of dried and powdered leaves were extracted with ethanol in Soxhlet apparatus for 24 hours (3 cycles hour). A dark brownish green colored residue was obtained after concentrating the extract under reduced pressure (Yield – 7.8%). The aqueous extract was obtained by macerating 500gms of powdered Leptadenia reticulata whole plant with 5 liters of distilled water (72 hours). The extract was filtered and concentrated under reduced pressure to obtain a green colored residue (Yield – 5.4%).

 

EVALUATION OF DIURETIC ACTIVITY:

Experimental Animals

The Wistar rats of either sex weighing 150-220g procured from the animal house were used for the study. The animals were maintained in polypropylene cages of standard dimensions at a temperature of 28±1° C and standard 12 hour: 12 hour day / night rhythm. The animals were fed with standard rodent pellet diet (Hindustan Lever Ltd) and water ad libitum. Prior to the experiment the animals were acclimatized to the laboratory conditions. All animals experiments conducted during the present study got prior permission from institutional animal ethics committee (IAEC) and followed the guidelines of IAEC.

 

DRUG: Furosemide tablet was collected from the local market of Narasapur was used as known diuretic agent. The standard solution was prepared by dissolving the tablet in the solvent.The dose of furosemide was maintained by 100mg/kg body weight.

 

Grouping of Animals

Animals were housed in a group of four in separate cages under controlled conditions of temperature (22 ± 2°C). All animals were given standard diet and water regularly. Animals were further divided in four groups with six animals in each group.

Group- I: Control , received in normal saline

Group-II:Standard(Furosemide100mg/kg,i.p),respectively in normal saline

Group- III: Aqueous extract of Leptadenia reticulata (100 mg/kg), respectively in normal saline

Group- IV: Ethanolic extract of Leptadenia reticulata (100 mg/kg), respectively in normal saline

PROCEDURE:

Lipschitz Method

Male rats (Wister albino strain) weighing 100 to 150gm were maintained under standard

Condition of temperature and humidity. 4 groups of six rats in each and were fasted and deprived of water for eighteen hours prior to the experiment. The first group of animals serving as control, received normal saline (25ml/Kg, p.o.) the second group received furosemide (100mg/Kg, i.p.) in saline; the third group received aqueous extract of ALR at the doses of 100 mg/Kg, respectively, in normal saline. the fourth group received ethanolic extract of ALR at the doses of 100 mg/Kg, respectively, in normal saline. The urine was collected in measuring cylinders up to 5 hrs after dosing. During this period, no food or water was made available to animals. The parameters taken for individual rat were body weight before and after test period, total concentration of Na+, K+, and Cl- in the urine. Na+, K+ concentrations were measured by Flame photometry and Cl- concentration was estimated by titration with silver nitrate solution (N/50) using three drop of 5% potassium chromate solution as indicator. Results are reported as mean ± SD, the test of significance (p<0.01 and p<0.05) was stastically16, 17. Furosemide sodium salt was given by stomach tube. Optimal dose activity relation was found to be 100mg/Kg of furosemide per kg body weight in series of supportive experiments. Results are reported as mean ± SD, the test of significance (p<0.01 and p<0.05) was stastically.

 

Statistical analysis

All the results are expressed as mean ± standard error. The data was analyzed statistically using ANOVA18 at a probability level of P < 0.001.

 

RESULTS AND DISCUSSION:

The preliminary phyto chemical analysis showed the presence of Flavanoids, Glycosides, Saponins, phenols, carbohydrates, Proteins and Amino acids, Tannins, Terpenoids and Alkaloids in both the extracts . Present study shows that the aqueous and ethanolic extracts of Leptadenia reticulata leave possess good diuretic activity. Urine volume, cation and anion excretion were increased, Na+/K+ ratio of 2.04 and 2.18 were obtained for aqueous and ethanolic extract respectively. The normal value for Na+/K+ ratio is reported to be 2.05 – 2.83. The concentration of aldosterone is found to be dependent on Na+/K+ ratio. If the Na+/K+ ratio falls below the normal in plasma the aldosterone secretion will be decreased and if the ratio rises above the normal value the aldosterone secretion will be increased. Significant increase in Na+, K+ and Cation excretion was observed in aqueous and alcoholic extract treated animals but it was less than the furosemide control. Further studies are required to assess the medicinal value of leaves of leptadenia reticulata as a potential diuretic agent (Table 1).

 

 

 

 

Table: 1 Diuretic activity of Leptadenia reticulata aerial parts extracts

S. No

Treatment

Dose

Mg/kg

No of rats

Urine volume(ml)

Electrolyte excretion

Na+

meq/100g

K+

meq/100g

Cl+

meq/100g

1

Control(normal saline)

25ml/kg

6

1.900 ±0.04

11.22±0.3

72.00±0.6

8.16±0.58

2

Furosemide

100

6

4.100±0.06

18.21±0.2

95.10±0.6

14.20±0.2

3

Aqueous

100

6

3.700±0.07

14.13±09*

85.12±0.0*

12.18±0.8

4

Ethanol

100

6

3.200±0.08

14.09±0.7

80.11±0.9

11.99±0.3

The values are expression of the mean standard error *P < 0.001vs.control

 

Diuretics relive pulmonary congestion and peripheral oedema. These agents are useful in reducing the syndrome of volume overload, decreases cardiac workload, oxygen demand and plasma volume, thus decreasing blood pressure19. Thus, diuretics play an important role in hypertensive patients. In present study, we can demonstrate that aqueous and alcoholic extract may produce diuretic effect by increasing the excretion of Sodium, Potassium and Chloride. The control of plasma sodium is important in the regulation of blood volume and pressure; the control of plasma potassium is required to maintain proper function of cardiac and skeletal muscles19. The regulation of Sodium, Potassium balance is also intimately related to renal control of acid-base balance. In present study aqueous and alcoholic extracts showed active principles such as flavanoids and terpenoids are known to be responsible for diuretic activity. Results of present investigation showed that both the aqueous and alcoholic is most effective in increasing urinary electrolyte concentration of all the ions i.e. Sodium, Potassium and Chloride. A primary law of the kidneys is that Na + excretion is a step function of mean arterial blood pressure (MABP) such that small increase in MABP cause marked increase in Na+ excretion20. One of the earliest strategies for the management of hypertension was to alter Na+ balance by restriction of salt in the diet. Diuretic agents having antihypertensive effects were used alone and had greater efficacy than all other antihypertensive drugs.

 

In this study pharmacological evaluation of diuretic action of aqueous and ethanolic extracts of Leptadenia reticulata was evaluated using furosemide under controlled laboratory condition. As diuretic therapy may lead to number of life threatening electrolytic disorder and toxicities, so safety profile studies are carried out following a sub chronic administration of extracts

 

CONCLUSION:

The results obtained in this study provide a quantitative basis to explain the traditional folkloric use of Leptadenia reticulata as a diuretic agent. This effect may be explored in the use of the plant in the management of some cardiovascular diseases.

 

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Received on 23.02.2012

Modified on 05.03.2012

Accepted on 18.03.2012                                               

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Research J. Pharmacology and Pharmacodynamics. 4(2): March - April, 2012, 84-86