Evaluation of cardiotonic activity of some traditionally used medicinal plants

K. Sanjeev Kumar, V. Vijay Kumar, A.G. Prasanthi, L.K. Kanthal, P. Bhoja Raju, K. Satyavathi*

Koringa College of Pharmacy, Korangi, Tallarevu (M), East Godavari Dist., Andhra Pradesh.

ABSTRACT:

The present study was undertaken to evaluate the cardiotonic activity of the aqueous, methanolic and chloroform extracts of the roots of Hemidesmus indicus and fruits of Terminalia bellirica, Terminalia chebula. The cardiotonic effect of aqueous, methanolic and chloroform extracts of the three plants was studied by using isolated frog heart perfusion technique (IFHP). Ringer solution without calcium was used as a vehicle for administration of aqueous, methanolic and chloroform extracts as test and digoxin as standard. A significant increase in the height of force of contraction (positive ionotropic effect) and decrease in heart rate (negative chronotropic effect) was observed and as compared to standard, test drug showed wide therapeutic index.

KEYWORDS: Cardiotinic activity, Frog heart perfusion technique, Digoxin, Therapeutic index, Hemidesmus indicus, Terminalia bellirica, Terminalia chebula.

INTRODUCTION:

Numbers of deaths in industrial world are increasing due to cardiac disease. Cardiac diseases are emerging as single largest contributors for morbidity in India. Cardiac glycosides and catecholamines are agents of choice in treatment of congestive cardiac failure (CCF)¹. But cardiac glycosides (e.g. digoxin) have narrow therapeutic index and hence cause many a times intoxication². Despite the advancement of knowledge in understanding the basic pharmacology of cardioactive drugs glycosides still have its adverse effects in terms of toxication³. Hence, there is a need for new drug research with wide therapeutic index and good cardiac activity, and by this aim, we have chosen Hemidesmus indicus, Terminalia bellirica and Terminalia chebula plants and evaluated its cardioactive potential.

Hemidesmus indicus- (sugandi, sariva, belonging to family: Asclepiadaceae(Apocynaceae)⁴, The flavanoid glycosides recognized in the flowers, were hyperoside, isoquercetin and rutin whereas in the leaves, only hyperoside and rutin were identified⁵. 2.5% of Tannins are present in leaves; roots are reported to contain sitoserol. A new ester identified include lupeol octacosanoate in addition to the known compounds viz., lupeol, amyrin, lupeol acetate, amyrin acetate, and hexatriacontane. Coumarins, triterpenoid saponins, essential oil, starch, tannic acid, triterpenoid saponins are also present. According to Ayurveda, root is cooling, aphrodisiac, antipyretic, alexiteric, antidiarrhoeal, astringent to bowels and useful in treatment of fevers, foul body odour, asthma, bronchitis, blood disorders, leucorrhoea, dysentery, diarrhoea, thirst, burning sensation, piles, eye troubles, epileptic fits, poisoning, rat bites etc. Ayurveda healers also prescribe it to men suffering from low libido and sexual impotence.

Terminalia bellirica-(beleric, bastard myrobalan) belonging to family: combretaceae⁶, Common Name(s):⁷ Arjuna, Axjun, Argun kahua, Kumbuk. Terminalia bellirica is a deciduous tree which is also grown as an avenue tree. The leaves are about 15 cm long petiolate, broadly and crowded towards the ends of the branches. The seeds contain several triterpenoids including β-sitosterol, and the saponin glycosides bellericoside and bellericanin. Other constituents include polyphenols (gallic acid, ellagic acid, phyllembin, ethyl galate, and chebulagic acid). Traditionally it is used as an astringent, laxative, infections of throat and chest. Terminalia bellirica has anti-inflammatory effect in the treatment of rheumatic disease. Dry fruit possesses potential broad-spectrum antimicrobial activity.

Terminalia chebula belonging to family combretaceae⁸, Common names: Haritaki, chebulic myrobalan. Terminalia chebula contains tannin, chebulic acid, glycosides, sugar, triterpenoids, steroids; small quantities of phosphoric acid. Major bioactive constituents are tannins, anthraquinones, chebulinic acid, chebulagic acid, chebulic acid, ellagic acid and gallic acid. Terminalia chebula is a rejuvenative, laxative (unripe), astringent (ripe), anthelmenthetic, nervine, expectorant, tonic, carminative, and appetite stimulant. It is useful for people who have leprosy (including skin disorders), anemia, narcosis, piles, chronic, intermittent fever, heart disease, diarrhea, anorexia, cough and excessive secretion of mucous, and a range of other complaints and symptoms.

MATERIAL AND METHODS:

· Drug: The fresh roots of Hemidesmus indicus, fruits of both Terminalia bellirica, Terminalia chebula were collected from Srisailam Reserve Forest, Kurnol Dist., Andhra Pradesh and authenticated in Department of Pharmacognosy, Koringa College of Pharmacy, Korangi, Andhra Pradesh.

· Animals: Frogs of Rana tagrina species⁹ were obtained from animal house of Korangi. Animals were feed with food and water adlibitum. They were maintained as per the norms of CPCSEA, and the experiment was conducted as per CPCSEA norms.

· Drug extract: Three different extractions are prepared for each plant material. 5gms of each plant material (Hemidesmus Indicus,Terminalia bellirica and Terminalia Chebula) was dissolved in 250ml of water, 250 ml chloroform, 250 ml methanol and kept a side for 24 hours with occasional shaking. After 24 hours each of extract was filtered and concentrated to a thick mass. This solid mass was used for screening of cardiotonic activity.

· Experimental methodology: ¹⁰ Pharmacological screening of cardiotonic activity of each extract using frog’s isolated heart by Isolated Frog Heart Perfusion technique.

Table-1: Effect of three plant extracts on isolated frog heart perfusion.

Sl. No.	Drug	Conc. (mg/ml)	Dose (in ml)	Heart Rate	HFC (mm)
*	Ringer	--------	Normal	55	03
1.	Aqueous extract of Hemidesmus indicus	1 mg/ml	0.1	53	05
2.			0.2	51	07
3.			0.3	50	08
4.	Chloroform extract of Hemidesmus indicus	1 mg/ml	0.1	54	07
5.			0.2	52	7.5
6.			0.3	50	08
7.	Methanolic extract of Hemidesmus indicus	1 mg/ml	0.1	49	06
8.			0.2	47	07
9.			0.3	45	8.5
10.	Aqueous extract of Terminalia bellirica	1 mg/ml	0.1	35	07
11.			0.2	33	08
12.			0.3	31	09
13.	Chloroform extract of Terminalia bellirica	1 mg/ml	0.1	52	06
14.			0.2	50	07
15.			0.3	49	7.5
16.	Methanolic extract of Terminalia bellirica	1 mg/ml	0.1	54	04
17.			0.2	53	05
18.			0.3	51	06
19.	Aqueous extract of Terminalia chebula	1 mg/ml	0.1	50	3.5
20.			0.2	49	04
21.			0.3	48	5.5
22.	Chloroform extract of Terminalia chebula	1 mg/ml	0.1	54	05
23.			0.2	52	06
24.			0.3	49	07
25.	Methanolic extract of Terminalia chebula	1 mg/ml	0.1	38	07
26.			0.2	36	8.5
27.			0.3	34	9.5
28.	Digoxin	0.5 mg/ml	0.1	58	07
29.			0.2	61	10
30.			0.3	63	--

HFC=Height of force of contraction.

The Statistical analysis was carried out using students’t test and it was found significant at P<0.001.

Evaluation of cardiotonic activity:

The frog of species Rana tigrina was pithed and pinned it to the frog board. A midline incision was given on the abdomen, the pectoral girdle was removed and the heart was exposed. The pericardium was carefully removed and put a few drops of hypodynamic frog ringer over the heart. The inferior venacava was traced, put a thread around it and given a small cut in order to insert the venous cannula. The cannula was inserted in the vein and the thread was tied to assure the cannula in place which is in turn connected to a saline bottle containing hypodynamic frog ringer solution^11,
12. A small cut in one of the aorta was given for the ringer to come out. Heart was isolated and attached to the stand with moderate flow of ringer. A thin pin hook was passed through the tip of the ventricle and with the help of a fine thread attached to the hook; it was tied to the free limb of the Sterling’s heart lever which was fixed to a stand. A proper tension was adjusted by altering the height of the lever. The normal heart rate was noted. All test samples were administered in different doses viz. 0.1ml, 0.2ml, 0.3ml respectively. The rate and force of heart contraction were noted as given in (Table-1 and Figure 1-9).

RESULT AND DISCUSSION:

All the dilutions of three plant extracts (Hemidesmus indicus roots, fruits of Terminalia bellirica and Terminalia chebula) restore cardiac activity of Hypodynamic frog heart i.e. it increases rapidity and force of contraction¹³.

The incremental dosage¹⁴ of three extracts produced positive inotropic and negative chronotropic effect on isolated frog heart and is dose dependent. The similar concentration of solution of digoxin produced positive inotropic and positive chronotropic effect. It was found that methanolic extract of Hemidesmus indicus, aqueous extract of Terminalia bellirica and methanolic extract of Terminalia chebula showed better response as compared to other extracts. It is interesting to know that three extracts have rapid onset of action compared to Digoxin. Further studies can confirm the reduced toxicity and this will be the advantage of three extracts over digitalis ^{15, 16}. Thus, in future it will be interesting to isolate the active chemical constituents which are responsible for the cardiotonic activity as well as to determine the possible mechanism of action.

REFERENCES:

1. Tripathi KD. Cardiac glycosides and drugs for CCF, Essentials of medical pharmacology. 5^th ed. Medical Publishers; 2003, 457-467.

2. Barar FSK, Essentials of Pharmacotherapeutics, 1st edition, S.Chand Publications, 1985, 253-254

3. Satoskar RS, Bhandarkar SD, Rege NN. Handbook of pharmacology and pharmacotherapeutics. 16^th ed. Popular Prakashan; 1997, 352-355.

4. Arun, V; Liju, V; Reena, J; et al. 2007. Traditional Remedies of Kani Tribes of Kottoor Reserve Forest, Agasthyavanam, Thiruvananthapuram, Kerala. Indian Journal of Traditional Knowledge, 2007, Vol.6(4), 589-594.

5. Alam MI, Audpy B, Gomes A. Viper venom neutralization by Indian medicinal plants (Hemidesmus indicus and Pluchea indica). Phytothery Resonance, 1996; 10: 58-61.

6. Alam Badrul . Antioxidant,Antimicrobial and Toxicity studies of the Different Fractions of Fruits of Terminalia belerica Roxb. Global Journal of Pharmacology. 2011;5(1):07-17.

7. Kumudhavalli MV, Vyas Mohit and Jayakar B. Phytochemical and Pharmacological evalution of the plant fruit of Terminalia belerica Roxb, International Journal of Pharmacy and Life Sciences. 2010; 1(1):1-11.

8. Suguna L, Singh S, Sivakumar P, Sampath P, Chandrakasan G. Influence of Terminalia chebula on dermal wound healing in rats. Phyther Res. 2002; 16:227–231.

9. Dama G.Y.,Tare H.L.,Gore M.S.,Shende V.S.,Deore S.R., Khandagale S.T.,Kandekar A.E. Comparative Cardiotonic Activity Of Helicteres Isora With Digoxin On Isolated Frog Heart. International Journal of Preclinical Research,Vol 2 | Issue 2 | Jul –Dec 2011 |81-86.

10. Mohire N.C., Salunkhe V.R., Bhaise S.B., Yadev A.V. Cardiotonic activity of aqueous extract of heartwood of Pterocarpus marsupium. Indian Journal of Experimental Biology, Vol.45, June 2007, 532-537.

11. Kulkarni SK, Handbook of Experimental Pharmacology, 2nd Edition, Vallabh Prakashan, 9, 1993, 74-76.

12. Kale SR, Kale RR. Practical Pharmacology and toxicology, 6th edition, Nirali Prakashan, 2003, 27- 28.

13. Tare HL, Thube BB. Comparative cardiotonic activity of C.angustifolia with digoxin on perfused frog heart. IJPRD, 1(1), 2009, 1-14.

14. Rai P.T. et al., Evaluation of Cardiotonic activity of leaves of Vitex negundo linn, International Journal of Green Pharmacy, Oct.-Dec.-2009, 306-309.

15. Goyal Ramesh K., Practicals in Pharmacology, 6th edition, B.S.Shah Prakashan, 2006-2007, 107-108.

16. Patel Rashmika P., Patel Manish P. Cardiotonic activity of isolated cardiac glycoside from the fruits of Corchorus aestuans linn, International Research Journal of Pharmacy, 2012, 3(7), 239-242.

Received on 29.01.2013

Modified on 10.02.2013

Accepted on 14.02.2013

Research J. Pharmacology and Pharmacodynamics. 5(2): March –April 2013, 87-91