Comparative Study of different Solvent Extract of Plectranthus amboinicus Stem for their Diuretic Activity

 

Noor Salma1*, Chandra Prakash K2, Syed Sagheer Ahmed3, Saba Tabassum4, Ahalya Devi K H5

1Department of Pharmacology, D R Karigowda College of Pharmacy, Hassan, Karnataka, India.

2,3Department of Pharmacology, Sri Adichunchanagiri College of Pharmacy,

Adichunchanagiri University, BG Nagara, Karnataka, India.

4Government Unani Hospital, Nagamangala, Karnataka, India.

5Department of Pharmacology, Bharathi College of Pharmacy, K.M Doddi, Karnataka, India.

*Corresponding Author E-mail: noorsalma.007@gmail.com

 

ABSTRACT:

Diuresis can be beneficial in removing toxins from the body. Globally traditional and folklore medicines plays a vital role in health care services. Plectranthus amboinicus belongs to Lamiaceae family is an Indian medicinal plant which has been utilized traditionally to treat various diseases. However, there is no systematic methodology for the use of Plectranthus amboinicus stem as a diuretic. Hence the present study is proposed to evaluate the diuretic activity of various solvent extract of Plectranthus amboinicus stem in the rat model. The acute toxicity study was performed in albino mice by fixed-dose method (OECD guidelines number 425) of CPCSEA. The diuretic activity was carried out using the Lipschitz method. The total urine volume collected in 24hours was observed. Sodium, potassium and chloride concentration in urine were also recorded. The ionic concentrations were recorded calorimetrically. Acute toxicity study proved no toxicity and mortality in any of the animal up to the dose of 2000mg/kg bodyweight. All three extracts of Plectranthus amboinicus have exhibited dose-dependent diuretic activity by increasing urine volume and excreting sodium ions in urine. The urine pH was also elevated. From the study, it can be concluded that the plant Plectranthus amboinicus could be considered as a diuretic agent. This current study encourages the use of this plant in folk medicine. Further study is required for isolation, characterization and formulation of the active constituents responsible for its diuretic activity.

 

KEYWORDS: Diuresis, Plectranthus amboinicus, Lipschitz method, petroleum ether, dichloromethane.

 

 

INTRODUCTION:

Diuretic action can be very beneficial, especially with regards to removing toxins from the body. Kidney helps to clean the blood by removing harmful substances from the body in the form of urine. Diuretic agents are valuable in several severe conditions like hypertension, Congestive Cardiac Failure, nephritis1, glaucoma and cirrhosis2. Globally traditional and folklore medicines plays a vital role in health care services3. In human beings, herbs are indicated as a main source of drugs in the management of various illnesses.

 

Across the globe, the utilization and acceptance of herbal remedies are in demand because of its safety4,5. As per the record of the World Health organization, about 80% of the world population is committed in traditional healing system6. In the current scenario, numerous Indian pharmaceutical industries are involved in the manufacturing of herbal products by virtue of favorable climate for cultivation7.

 

Plectranthus amboinicus belongs to the family Lamiaceae, commonly known as Country borage in English8. It is grown as a household herb in Tamilnadu. This herb is native to East Indies and is widely cultivated in Africa and almost all tropical countries9. The leaves of this herb were used traditionally in the treatment of sore throats, cough, asthma, malarial fever, nasal congestion and cancer. It is also utilized as a remedy for rheumatism, infections and flatulence. It is also having hepatoprotective, diuretic, antimalarial, antibacterial, antiepileptic and antioxidant property10,11. Various activities like antifungal, anti-inflammatory12, antianxiety13, antibiotic14, antimicrobial15, hepatoprotective16, anticancer17, anthelmintic, bronchodilator18 and antidiabetic19 activities were reported for this plant. There is no proper systematic evidence for the diuretic activity of Plectranthus amboinicus stem. Hence the present study is proposed to evaluate the diuretic activity of various solvent extract of Plectranthus amboinicus stem in the rat model.

 

MATERIALS AND METHODS:

Extraction:

Plectranthus amboinicus plant was collected locally from Nagamangala Taluk. The powdered stem was extracted by soxhlet apparatus using three solvents (petroleum ether, dichloromethane and ethanol) based on their polarity. These extracts were stored in a required temperature (40C) until used for the experimental studies.

 

Experimental Animals:

Albino mice and rats weighing around 140 to 200g and 20 to 25g respectively were procured and maintained under standard environmental conditions. Animals were served with free access to food and water. Approval was taken from the Institutional Animal Ethical Committee and received ethical clearance number (SACCP-IAEC/28/2015).

 

Acute toxicity studies:

Plectranthus amboinicus extracts were tested for acute toxicity using albino mice as per OECD guidelines 425. Animal’s survival was noted for 48 hours and 14 days after administration of the different doses of extracts.

 

Phytochemical Screening:

Presence of Phytoconstituents was proved by performing standard procedures given in the textbook of Kokateand Khandelwal followed by some modifications20-23

 

Diuretic Activity:

The method of Lipschits et.al., was employed for the assessment of diuretic activity. 48 albino rats of either sex weighing between 140-200gm were selected and divided into 8 groups of 6 animals in each group. On the day of the experiment, animals were given 1ml/200gm of body weight normal saline orally. The first group was served as control (receive vehicle only); the second group had received 100mg/kg of furosemide per oral route (standard drug). Third to eight groups of the animals were given with all the solvent extracts of Plectranthus amboinicus at a dose of 200 and 400mg/kg body weight.

 

Immediately after the dosing, all the rats were housed in the metabolic cages with special provision for urine and faeces collection. Animals were kept at room temperature of 35±10C throughout the experiment. The urine was collected in a measuring cylinder for the period of 24 hours. During this phase, food and water were withdrawn from all the animals. The total volume of urine collected in 24hours and various ionic concentrations in the urine like Na+, K+, Cl- were noted. The concentration of sodium and potassium ions was measured calorimetrically and the results were reported as mean± SEM24,25

 

Statistical Analysis:

The results were stated as Mean±SEM and considered to statistical analysis by ANOVA followed by Dunnett ‘t’ test.

 

RESULTS:

The Preliminary phytochemical investigation stated the occurrence of flavonoids, alkaloids, carbohydrates, proteins, triterpenoids, tannins and glycosides in the ethanolic extract. In dichloromethane extract, various constituents like carbohydrates, saponins, proteins, glycosides, steroids and alkaloids were present and in petroleum ether extract saponins and steroids were observed.

 

The petroleum ether, dichloromethane and ethanol extracts did not caused any toxicity or mortality in any of the mice when administered at a dose of 2000mg/kg body weight orally. To study diuretic activity the extracts were administered to the rats in the dose of 200 and 400mg/kg body weight which is equal to 1/10th and 1/5th of maximum safe dose respectively.

 

Diuretic activity:

All the three extracts of Plectrantus amboinicus showed dose-dependent diuretic activity by increasing urine volume and excreting Sodium (Na+), Potassium (K+) and Chloride (Cl-) ions. The detailed activity is represented in table 1.

 

Table 1: Diuretic activity of Stem extracts of Plectranthus amboinicus in experimental rats.

Group

Volume of urine in ml

Na+ mmol/L

K+ mmol/L

Cl- mmol/L

PH

Control

3.08 ± 0.15

47.66 ± 0.95

47.5 ± 1.25

75.83 ±0.60

5.61± 0.11

Standard

8.63 ± 0.17**

99.5 ± 1.33**

55.83 ± 1.01**

112.16 ± 1.30**

7.31 ± 0.04

PEPA 400

5.55 ± 0.19**

70.5 ± 0.76**

51.66 ± 1.22*

83.33 ± 0.61**

6.35 ± 0.04

PEPA 200

4.06 ± 0.12**

62.5 ± 1.08**

48.16 ± 0.94ns

62.83 ± 0.60**

6.05 ± 0.06

DCMPA 400

4.88 ± 0.13**

69.16 ± 0.87**

52.66 ± 0.55**

66.66 ± 0.80**

6.33 ± 0.20

DCMPA 200

3.93 ± 0.24**

61.83 ± 0.90**

48.83 ± 1.04ns

56.66 ± 0.49**

5.85 ± 0.06

ALPA 400

6.58 ± 0.15**

91.16 ± 1.70**

54.66 ± 0.88**

94.83 ±0.90**

7.03 ± 0.06

AL PA 200

5.51 ± 0.10**

72.16 ± 1.01**

49.5 ± 0.67ns

85.66 ± 1.28**

6.73 ± 0.09

Values are expressed as mean ± SEM (n = 6). ***P<0.001, **P< 0.01 and *P< 0.05 ns = not significant

 

PEPA 400: Petroleum ether extracts of Plectranthus amboinicus (400mg), PEPA 200: Petroleum ether extracts of Plectranthus amboinicus (200mg), DCMPA 400: Dichloromethane extracts of Plectranthus amboinicus (400mg), DCMPA 200: Dichloromethane extracts of Plectranthus amboinicus (200mg), EEPA 400: Ethanolic extracts of Plectranthus amboinicus (400 mg), EEPA 200: Ethanolic extracts of Plectranthus amboinicus (200mg).

 

DISCUSSION:

Plectranthus amboinicus is a herb being used in folk medicine for treating many ailments. Diuresis has the key property of extracting harmful substances and filtering blood through kidneys and thus improves the functioning of the kidneys. The diuretic action occurs due to the stimulation of the urinary tract by triggering neurohumoral pathway which may be due to the effect of electrolytes on the renal epithelium26. Hypertension has a chief role in the progression of cerebrovascular disorders, ischemic heart disease, other heart-related problems and renal failure. Management of hypertension may reduce the incidence of stroke and myocardial infarction up to 40% and 15% respectively.

 

Diuretics are used effectively in conjunction with other class of drugs to treat hypertension. A diuretic agent diminishes the vascular preload by lowering the volume of circulating intravascular fluid. It is also having the ability to reduce capillary and venous pressure by its vasodilatation action, thereby improves cardiac functioning.27 There is a necessity of plant-based novel diuretics with no or negligible adverse effects. This would be the best alternative to synthetic drugs because of its similar beneficial effect with least side effects. Various Phytoconstituents like flavonoids, saponins, volatile oils, sterols and triterpenes are known to posse’s diuretic activity.28,29 Similar constituents are present in Plectranthus amboinicus. Thus, the diuretic activity of this herb is due to the presence of these constituents which provide a systematic background for the traditional use of this plant for its diuretic potential. The present study concludes that the effects of petroleum ether, dichloromethane and ethanolic stem extracts of Plectranthus amboinicus exhibited dose-dependent diuretic activity. Further study concerning the isolation, characterization and formulation of the active principles responsible for diuretic activity at the cellular level is required.

 

ACKNOWLEDGEMENT:

The authors would like to extend a heartfelt thanks to the Principal and management of Sri Adichunchanagiri College of Pharmacy for facilitating this research.

 

CONFLICT OF INTEREST:

Declared None.

 

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Received on 19.06.2020           Modified on 13.08.2020

Accepted on 28.09.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(9):4727-4730.

DOI: 10.52711/0974-360X.2021.00822