A comparative evaluation of cardioprotective activity of...

Indian Journal of Natural Products and Resources

Vol. 2(3), September 2011, pp. 335-344

A comparative evaluation of cardioprotective activity of two Makandi

(Coleus forskohlii Willd.) formulations against isoproterenol induced

myocardial infarction in hyperlipidaemic rats

Madhavi Jagtap1, B K Ashok

2, Sulakshan S Chavan

3, H M Chandola

1 and B Ravishankar

4*

1Department of Kayachikitsa, 2Pharmacology Laboratory, 3CCRAS, Pharmacological Research Unit, Institute for

Post Graduate Teaching & Research in Ayurveda, Gujarat Ayurved University, Jamnagar - 361 008, Gujarat, India 4SDM Research Centre for Ayurveda and Allied Sciences, Kuthpady, Udupi - 574 118, Karnataka, India

Received 21 July 2010; Accepted 29 March 2011

The present study was undertaken to evaluate cardioprotective activity of Makandi (Coleus forskohli Willd.) Churna and

Ghanavati on the basis of electrocardiographic, biochemical, cardiac output and histopathological parameters against

isoproterenol (ISO) induced myocardial infarction in diet induced hyperlipidaemic rats. The drug treatment and

hyperlipidaemic diets were administered for 20 consecutive days. Myocardial injury was induced by injecting ISO

(85 mg/kg) to rats at an interval of 24 h for two days. Forty eight hours after the first dose of ISO injection, parameters like

ECG, cardiac output, biochemical and histological observations of the heart tissues were performed. Hyperlipidaemic diet

followed by ISO significantly increased heart weight, altered serum lipid profiles, SGPT and ALP activity, caused ST

segment elevation, prolonged QT interval and QTc in ECG, increased blood pressure and decreased cardiac output.

Histopathologically also, heart showed severe cytoarchitectural disturbances like myonecrosis, fatty changes and

endocardial oedema. When administered orally, both the formulations of Makandi decreased atherogenic index, almost

normalized ST segment elevation, QT interval prolongation and cardiac output. Histopathologically also remarkable

protection was observed. Analysis of the results showed that Makandi in Churna form has both anti-atherogenic potential

and cardioprotective activity, while Ghanavti has only weak cardioprotective activity.

Keywords: Cardioprotective, Coleus forskohlii, Hyperlipidaemia, Makandi, Makandi churna, Makandi ghanavati,

Myocardial infarction.

IPC code; Int. cl. (2011.01) A61K 36/53, A61K 125/00, A61P 9/00

Introduction

Coleus forskohlii Willd. is popularly known as

Makandi in Ayurveda belonging to the family

Lamiaceae, has been used in traditional medicine

since ancient times for treatment of heart diseases,

abdominal colic and respiratory disorders1,2

. The plant

is a rich source of a diterpene, forskolin which is

virtually responsible for all reported pharmacological

activities. It is an activator of adenylate cyclase3,4

and also has positive ionotropic, anti-glaucoma,

anti-inflammatory, anti-platelet aggregation,

bronchospasmolytic actions5 and reduce body weight

by increasing lean body mass6. However, almost all

activities have been carried out on active principle

forskolin i.e., on the isolated fraction of the drug. Till

date no work has been reported on conventional

Ayurvedic preparations of this drug.

The majority of the studies show that

hyperlipidaemia, independently from the development

of coronary atherosclerosis, worsens the outcome of

ischaemic injury7. These findings emphasize the

necessity of lipid lowering therapy and promote the

development of new cardioprotective drugs that are

capable to reverse the increased susceptibility of

hearts to ischaemic stress and to recapture cardiac

stress adaptation in hyperlipidaemia. Thus, the present

study was undertaken to evaluate comparative

cardioprotective effect of two formulations prepared

from Makandi, viz. Makandi churna (powder of

roots) and Makandi ghanavati (concentrated form of

aqueous extract from roots) against isoproterenol

induced cardiac injury in hyperlipidaemic diet

induced hyperlipidaemic rats.

__________

*Correspondent author: E-mail: [email protected];

Phone: 09483929319 (Mob.)

INDIAN J NAT PROD RESOUR, SEPTEMBER 2011

336

Materials and Methods

Test formulations

The fresh tuberous roots of Makandi (C. forskohli,

Plate 1) were procured from local market and

authenticated by qualified pharmacognosist of our

institute through macroscopical and microscopical

characters. The tuberous roots were washed with

water, cut into pieces and dried under shade. The

Churna (MC) and Ghanavati of Makandi (MG) were

prepared from dried roots as per classical procedure8,9

.

Prelimnary phytochemical studies

Both Churna and Ghanavati of Makandi were

subjected to preliminary phytochemical studies

including quantitative phytochemical investigation

through HPLC analysis for important chemical

moiety as forskolin10

. The separation was carried out

on a C-18 column (Inertsil C18 250 mm × 4.6 mm i.d.

with a 5 µm particle size) using Acetonitrile : Water

(90:10 v/v) as mobile phase. The eluent was

monitored using a UV-Visible detector at 210 nm.

Standard Forskolin (87.00 %) was used as marker

compound.

Animals

Wistar strain albino rats of either sex weighing

200 ± 30 g were selected from the animal house

attached to the institute. They were housed at

25 ± 03°C with constant humidity of 50-70% on a

12 hour natural day and night cycles. They were fed

with diet Amrut brand rat pellet food supplied by

Pranav Agro Industries, Baroda and tap water was

given ad libitum. Institutional Animal Ethics

Committee had approved the experimental protocol

(Approval number IAEC/09-10/05MD 04) and the

care of animals was taken as per the CPCSEA

guidelines.

Dose selection and schedule

The clinical dose of Makandi ghanavati and

Makandi churna are 3 and 4.2 g, respectively10

. The

dose for experimental animals was calculated by

extrapolating the human dose to animals (270 mg

rounded to 300 mg/kg and 378 mg rounded to

400 mg/kg, respectively) based on the body surface

area ratio by referring to the standard table of Paget

and Barnes (1969)11

. The human doses of the

formulations were decided on the basis of the clinical

experiences, palatability, since the majority of clinical

studies have used injected forskolin, it is unclear if

oral ingestion of these formulations will provide

similar benefits in the amounts recommended by

various authors [About 80 g of dried course powder of

Makandi gives 14 g of Ghana (aqueous extract)]. The

drug solutions were made with distilled water and

administered to animals (0.5 ml/100 g body weight)

with the help of gastric catheter sleeved to syringe.

The drugs were administered to over night fasted

animals.

Statistical analysis

The results were presented as Mean ± SEM for six

rats in each group. Statistical comparisons were

performed by both unpaired student’s t test and one

way ANOVA with Dunnets’ multiple t test as post-hoc

test by using Sigma stat software (version 3.1) for

all the treated groups with the level of significance

set at P<0.05.

Plate 1 Coleus forskohlii: a-Plant, b-Tuberous roots

JAGTAP et al.: EVALUATION OF CARDIOPROTECTIVE ACTIVITY OF MAKANDI FORMULATIONS

337

Experimental protocol

The experimental rats were divided into six groups

of 6 animals each and treated as follows:

Group I: Normal control rats received distilled

water (NC)

Group II: Cholesterol control rats treated with

hyperlipidaemic diet (CC)

Group III: Isoproterenol control rats treated with

Isoproterenol (ISO) (85 mg/kg body weight,

s.c. in saline).

Group IV: Cholesterol plus Isoproterenol

(CCISO) control group

Group V: Makandi ghanavati (MG) +

cholesterol + Isoproterenol (MG + CCISO)

Group VI: Makandi churna (MC) + cholesterol

+ Isoproterenol (MC + CCISO)

Test drugs and vehicles were administered to

respective groups at morning hours and continued for

20 days. The hyperlipidaemic diet was administered

to different groups to induce hyperlipidaemia based

on previous studies12,13

. The hyperlipidaemic diet

includes hydrogenated vegetable oil (Vanaspati Ghee-

'Raag' brand, Batch No. BA 55, Adani Wilmar Ltd.,

Gujarat) and cholesterol extra pure powder (Batch

No. 14022 Suvidhnath Laboratories, Baroda) made in

to 20% suspension in coconut oil (Parachute coconut

oil, Batch No. PSO73, Goa). The suspension was

administered (0.5 ml/100 g) to the rats daily for

20 days (at evening hours) to all the groups except

normal control (Group I) and isoproterenol control

(Group II) group. On 20th day, 1 hour after the

administration of test drug and vehicle, ISO was

administered subcutaneously. Two doses of ISO

(85 mg/kg) were administered at 24 hours interval14

.

Measurement of ECG

At the end of experimental period (after 24 h of

second ISO injection) the rats were anaesthetized with

ether and ECGs were recorded using a portable

electrocardiogram machine (Cardiofax-Medicaid

systems). ECG was recorded by using only the four

standard (limb) leads attached to four extremities of

the animals, the chest leads were not used. In all cases

of myocardial infarction, Lead II show the clear,

distinct individual waves than Lead I and III.

Therefore, ECG was monitored on Lead II only.

Measurement of blood pressure and cardiac output

Systolic blood pressure, diastolic blood pressure,

mean blood pressure, pulse rate, cardiac output (CO),

left cardiac work (LCW), left ventricular ejection time

(LVET), pre-ejection period (PEP) and stroke index

(SI) were recorded using impedance cardiographic

analysis (ICG) provided with BIOPAC student

lab system using Sramek/VEPT method in

anaesthetized rats. Biochemical analysis

After recording the BP and cardiac output, blood

was collected from retro-orbital plexus, serum was

separated and used for estimation of different serum

biochemical parameters. The procedure followed for

this was a requisite quantity of serum fed to the auto

analyzer (Fully automated Biochemical Random

Access Analyzer, BS-200; Lilac medicare Pvt. Ltd.,

Mumbai) which was automatically drawn in to the

instrument for estimating different biochemical

parameters like serum total cholesterol (CHOD–PAP,

end point)15

, serum HDL cholesterol (Trinder

reaction)16

, serum triglyceride (GOP–PAP method,

end point)17

, serum alkaline phosphatase (IFCC

Method, Kinetic method)18

, SGOT (IFCC method

without Pyridoxal Phosphate)19

and SGPT activity

(IFCC method, kinetic without Pyridoxal

Phosphate)20

, VLDL was calculated by using formula

VLDL=TG/5, LDL by LDL={TC-(HDL-VLDL)}21

and atherogenic index by=TC/HDL22

. Histopathological studies

At the end of the study, all the rats were sacrificed

by overdose of ether anaesthesia and the heart was

dissected out, washed in ice cold saline. The weight of

heart was noted and the tissues were immediately

fixed in 10% buffered neutral formalin solution. After

fixation, tissues were embedded in paraffin and serial

sections were cut and each section was stained with

hematoxylin and eosin23

. The slides were viewed

under Trinocular Research Microscope (Germany) at

various magnifications to note down the changes in

the microscopic features of the tissues studied.

Results and Discussion

Effect on heart weight

The heart weight of rats treated with isoprotrenol

significantly increased when compared with the

normal control rats (Table 1). Nirmala and

Puvanakrishnan (1996)24

have reported that this

increase in heart weight might be due to increased

oedema within intramuscular space, massive vascular

hemorrhage and/or extensive necrosis of cardiac

muscle fibres followed by the invasion of damaged

tissues with inflammatory cells. These observations


338

are in line with the histopathological findings of our

study. Both the test drugs did not attenuate the

observed cardiac weight gain. This may be due to

persistent oedematous condition in the heart in spite

of drug treatment.

Effect on serum lipid profiles

High cholesterol diet is regarded as an important

factor in the development of cardiac diseases

since it leads to development of hyperlipidemia,

atherosclerosis and ischemic heart disease25

. In the

present study feeding of hyperlipidemic diet not only

elevated serum cholesterol level but also elevated BP.

Injection of isoprenaline in hyperlipidemic diet given

rats produced higher degree of cardiac injury. Though

the injury profile was similar to isoprenaline treated

but the severity of the injurious lesions were

remarkably high (Plate 2d).

Administration of cholesterol and hyperlipidemic

diet lead to significant elevation of serum total

cholesterol, LDL cholesterol and atherogenic index.

The increase in total cholesterol was maintained even

after injection of the isoprenaline, in addition

significant decrease in serum triglyceride, HDL-

cholesterol and VLDL-cholesterol was observed.

Both MG and MC failed to attenuate serum lipids

elevations. However, CCISO induced atherogenic

index which is the ratio of total cholesterol to

HDL-cholesterol, was remarkably reversed by both

MG and MC treatment (Table 2).

The serum triglyceride levels were found

moderately lowered in CC and ISO given groups.

When both of them were combined as in case of

CCISO group, significant lowering of triglyceride

level was observed. This lowering can be attributed to

isoprenaline induced modulation of triglyceride

metabolism possibly through increased utilization.

Since similar decrease was observed in case of VLDL

cholesterol also. It implies that metabolism of VLDL

may also be modulated. The probable mechanisms of

the observed triglyceride lowering can be due to

increase in β-oxidative degradation of fatty acids, and

lipoprotein lipase activity leading to increased

metabolism of triglycerides in VLDL particles and

chylomicrons and diversion of fatty acids to muscles

for utilization in their activity along with reduction in

Table 1 Effect of Makandi ghanavati and Makandi churna on heart weight

Parameter NC CC ISO CCISO MG + CCISO MC + CCISO

Heart weight

(g/100g)

0.325 ± 0.0169 0.312 ± 0.0098 0.453 ±

0.0144***®

0.436 ±

0.0152***®

0.423 ± 0.0114 0.454 ± 0.0267

The data were expressed as Mean ± SEM; significant differences in each group vs the control is ***P< 0.001.

(® ONE WAY ANOVA-F value 27.133; P<0.001 DMTT- P<0.05 for ISO and CCISO vs Control).

Table 2 Effect of Makandi ghanavati and Makandi churna on serum lipid profiles


Serum

Cholesterol

64.00 ± 2.37 73.50 ± 3.09* 82.50 ± 8.61* 81.00 ± 4.76** 98.33 ± 7.81 86.50 ± 8.01

Serum HDL

cholesterol

36.33 ± 3.04 30.00 ± 1.84 25.17 ± 2.01* 26.83 ± 1.70* 34.33 ± 2.20 36.50 ± 3.27#®®

Serum VLDL

cholesterol

20.57 ± 2.75 17.30 ± 1.21 12.87 ± 3.02 11.60 ± 1.12* 10.00 ± 1.37 14.10 ± 2.76

Serum LDL

cholesterol

48.23 ± 2.53 65.40 ± 6.26* 70.20 ± 7.60* 67.77 ± 5.06** 74.00 ± 7.99 64.10 ± 6.60

Atherogenic

index

1.82 ± 0.15 2.68 ± 0.19**® 3.43 ± 0.50**® 3.42 ± 0.32*** ®

2.28 ± 0.29#¥ 2.42 ± 0.31#

Serum

triglyceride

102.83 ± 13.75 86.50 ± 6.07 64.33 ± 15.09 58.00 ± 5.59* 50.00 ± 6.85 70.50 ± 13.82

The data were expressed as Mean ± SEM and unit of each parameter is (mg/dL).

Significant differences in each group vs. the normal control are *P< 0.05, **P< 0.01 and ***P< 0.001; vs CCISO are #P <0.05

(® ONE WAY ANOVA-F value 5.726; P=0.005 DMTT- P<0.05 for ISO and CCISO vs Control)

(®®ONE WAY ANOVA-F value 4.982; P=0.025 DMTT- P<0.05 for MC + CCISO vs CCISO)

(¥ONE WAY ANOVA-F value 4.240; P=0.038 DMTT- P<0.05 for MG + CCISO vs CCISO)


339

hepatic production of VLDL26

. CCISO induced

lowering was not antagonized by the test preparations.

Instead, further lowering was observed. The reason

for this lowering of VLDL was not known.

Effect on serum enzymes

Highly significant elevation in both SGOT and

SGPT activity was observed in CCISO treated rats

(Table 3) in comparison to control group. The

increase was 182% in case of SGPT and 138% in case

of SGOT. In MG treated CCISO receiving group,

slightly higher elevation of SGPT activity was

observed where as in MC treated CCISO receiving

group only 47% inhibition was observed. This

indicates marked reversal of CCISO produced

elevation. In case of SGOT activity, in MG treated

CCISO receiving group, a moderate 86.75% increase

was observed which was moderately less in

comparison to CCISO control. In MC treated CCISO

receiving group, no decrease was observed. The

observed elevation correlates very well with the

histopathological changes observed in heart of CCISO

combination group indicating the observed elevation

is due to drastic tissue damage caused to heart.

Though perfect correlation was not obtained in drug

treated group the observed decrease indicates that

both of them have attenuating effect on CCISO

induced tissue injury.

Significant elevation of serum ALP activity was

observed in CC and CCISO control group. In MG

treated group a further elevation was observed where

as in MC treated group a moderate fall of 24% was

observed in CCISO control group. This indicates that

MC treated CCISO receiving group has moderate to

good reversal effect on the CCISO induced toxic

effect probably involving bone and cardiac tissue.

Effect on ECG parameters

Electrocardiograph abnormalities are the main

criteria generally used for the definite diagnosis of

myocardial infarction27

. ECG changes were derived

from standard II-lead ECG and the information

recorded are PR, QRS, QT and RR intervals, heart

rate and QTc interval (Table 4, Fig. 1). The main

changes observed were elevation of ST segment

(Fig. 1d), prolongation of QT interval and QTc

interval in CC and CCISO control group in

comparison to normal control group. QT interval

provides a measure of ventricular repolarization and is

determined by the balance of the repolarizing inward

sodium and calcium currents, and the outward

potassium and chloride currents28

. The QT interval

correlates with measurements of cardiac autonomic

function, with cardiac vagal dysfunction resulting in

prolongation of the QT interval. QT interval

represents both the dispersion and the lengthening of

the AP duration and correlates with the LV mass. The

prolongation of this interval is considered as an

important parameter to determine cardiac toxicity

potential29

. In humans and large animals, QT interval

varies strongly and inversely with heart rate.

Consequently, in clinical practice and human

pharmacologic studies it is typical to correct the

measured QT interval for heart rate in order to

obtain measures of QT interval which are heart

rate independent (QTc). The prolongation of both

these parameters in CC and CCISO control group

indicates presence of cardiac injury and aberration in

the ventricular repolarization. The fact that both the

test formulations were effective in reversing this

CCISO induced QT interval prolongation points

towards the presence of significant cardioprotective

activity in them.

Effect on blood pressure

Rats in CC group showed significantly elevated blood pressure in terms of systolic, diastolic and mean blood pressure in comparison to normal control rats (Table 5). Administration of ISO and CCISO leads to

Table 3 Effect of Makandi ghanavati and Makandi churna on serum enzymes


SGPT

activity (IU)

49.17 ± 7.77 66.29 ± 10.20 94.60 ± 17.78* 136.20 ± 16.03*** 122.17 ± 15.15** 080.50 ± 12.12#

SGOT

activity (IU)

186.50 ± 13.53 190.57 ± 22.55 196.80 ± 33.96 297.17 ± 32.26** 277.33 ± 24.46 337.50 ± 68.02

Alkaline

phosphatase

activity (IU/L)

136.83 ± 39.68 301.67 ± 30.42** 223.67 ± 8.74* 276.17 ± 46.34* 323.83 ± 41.63** 208.50 ± 42.19

The data were expressed as Mean ± SEM. Significant differences in each group vs the normal control are *P< 0.05, **P< 0.01 and

***P< 0.001; vs CCISO is #P <0.05.


340

moderate non-significant fall in BP in comparison to normal control. This shows that ISO instead of elevating BP has the tendency of lowering it in CC given groups. Both MG and MC failed to antagonize the hyperlipidaemic diet induced BP elevation.

Effect on cardiac output parameters

The main and significant change that was observed

after feeding rats with cholesterol rich diet and

CCISO combination was decrease in cardiac output

(Table 6). This decrease was found completely

reversed in MC treated group. MG did not reverse this

decrease, however a slight further decrease was

observed. The left cardiac work was also found to be

decreased in CCISO group in comparison to normal

control rats. This decrease was also reversed by MC

while MG did not produce the reversal. Left

ventricular ejection time was found to be moderately

prolonged by CCISO combination this moderate

prolongation was not influenced by either of the test

drugs. Stroke index was found moderately decreased

in CCISO group in comparison to normal control. The

stroke index was found doubled in MC administered

group in comparison to CCISO control whereas in

MG treated group, a marginal decrease was observed.

Thus, the overall profile indicates that administration

of CCISO combination leads to significant reduction

in the working capacity of the heart. This

incapacitation was completely reversed by MC while

MG was not effective. Histopathological findings

No inflammatory cells infiltration was seen in the

heart of normal control rats (Plate 2a & b).

Isoprenaline injection to rats leads to significant

myonecrosis, infiltration of inflammatory cells, fatty

changes and endocardial oedema compared to normal

control (Plate 2c). In CCISO control group these

Table 4 Effect of Makandi ghanavati and Makandi churna on ECG parameters


PR interval 0.035 ±

0.0034

0.040 ±

0.0022

0.041 ±

0.0051

0.038 ±

0.0016

0.036 ±

0.0036

0.037 ±

0.0019

QRS interval 0.043 ±

0.0021

0.046 ±

0.0015

0.040 ±

0.0039

0.046 ±

0.0029

0.035 ±

0.0056

0.046 ±

0.001

QT interval 0.082 ±

0.0030

0.102 ±

0.0031***

0.109 ±

0.0179

0.113 ±

0.0078***

0.0872 ±

0.0042##

0.0908 ±

0.0025#

Heart rate 339.111 ±

11.08

354.667 ±

39.95

362.200 ±

44.52

384.200 ±

22.24

418.833 ±

12.19

463.750 ±

27.29

RR interval 0.177 ±

0.0062

0.181 ±

0.0206

0.178 ±

0.0258

0.158 ±

0.0086

0.144 ±

0.0043

0.131 ±

0.0075

QTc 0.193 ±

0.0056

0.245 ±

0.0093***®

0.255 ±

0.0183**®

0.285 ±

0.0109***®

0.231 ±

0.0110#β

0.252 ±

0.0068#

The data were expressed as Mean ± SEM.

Significant differences in each group vs. the control are *P< 0.05, **P< 0.01 and ***P< 0.001; vs CCISO are #P<0.05 and ##P<0.01.

(® ONE WAY ANOVA-F value 10.030; P<0.001 DMTT-P<0.05 for CC, ISO and CCISO vs Control).

(β ONE WAY ANOVA-F value 7.408; P=0.006 DMTT-P<0.05 for MG + CCISO vs CCISO).

Table 5 Effect of Makandi ghanavati and Makandi churna on blood pressure


Systolic BP

(mm of Hg)

127.00 ± 09.29 157.0 ± 04.69* 113.400 ± 11.59 130.667 ± 06.87 132.00 ± 06.25 148.75 ± 03.54

Diastolic BP

(mm of Hg)

83.000 ± 05.71 102.16 ± 04.54* 073.800 ± 06.95 093.667 ± 05.51 95.600 ± 04.55 110.250 ± 07.37

Mean blood

pressure

(mm of Hg)

101.167 ± 09.55 128.667 ± 04.08* 93.800 ± 08.89 112.333 ± 06.02 114.800 ± 04.15 128.500 ± 04.94

The data were expressed as Mean ± SEM.

Significant differences in each group vs the control is *P< 0.05.


341

Fig.1 Photographs of ECG leads showing: a-normal ECG pattern of control (NC) group; b-slight elevation of ST segment in CC

group; c-elevation of ST segment in ISO group; d-marked elevation of ST segment in CCISO group; e-slight elevation of ST segment in

MG treated group; f-elevation of ST segment in MC treated group


342

pathological features were found further increased

involving vast area of the myocardium (Plate 2d).

This shows injection of ISO to hyperlipidemic

diet given rats further enhances myocardial injury.

Administration of MG moderately stemmed these

CCISO combination induced myocardiopathy

(Plate 2e). MC provided marked protection against

CCISO induced myocardiopathy (Plate 2f). Thus,

Plate 2 Photomicrographs of heart sections (1× 400 magnification): a-control group (NC) showing normal cytoarchitecture; b-CC

group showing mild fatty changes and myocarditis; c-ISO group showing multiple focal necrosis with infiltration of leukocytes (arrow);

d-CCISO group showing multiple focal necrosis with infiltration of leukocytes (arrow) and proliferating fibroblasts; e-MG treated group

showing very less degree of focal necrosis and infiltration of leukocytes; f-MC treated group showing almost normal cytoarchitecture.


343

Makandi in the form of Churna may be of therapeutic

and prophylactic value in the treatment of myocardial

infarction.

Myocardial infarction is the rapid development of

myocardial necrosis caused by critical imbalance

between the oxygen supply and the demand of the

myocardium. A better understanding of the processes

involved in myocardial infarction has stimulated the

search for new drugs which could limit the

myocardial injury30

. The major abnormalities noticed

in myocardial infarction are lipidaemia, peroxidation

and loss of plasma membrane integrity31

. Further,

studies indicate that South Asians have elevated levels

of LDL cholesterol and triglycerides, while also

suffering from a deficiency in HDL cholesterol32,33

.

The mechanism by which hyperlipidaemia may

influence the severity of myocardial ischaemia is not

exactly known, however, accumulation and

redistribution of tissue/membrane cholesterol and the

resulting changes in sarcolemmal and mitochondrial

membrane micro-viscosity rather than the direct effect

of high serum lipoprotein levels and coronary

atherosclerosis may account for this34

. A decrease in

cardiac NO bioavailability35

and ecto-5'-nucleotidase

activity, an inhibition of the mevalonate pathway36

,

as well as enhanced apoptotic cell death have been

also shown to contribute to increased

ischaemia/reperfusion injury and loss of

preconditioning in hyperlipidaemic animal models37

.

Conclusion

Based on activity profile obtained, it can be

concluded that both the formulations of Makandi have

good anti-atherogenic potential and the Churna

formulation has remarkable cardioprotective activity,

while Ghanavati has weak cardioprotective activity. It

is more interesting to note the observed differences in

the activity profiles of the formulations, despite the

fact that both possess the same active ingredient. The

HPLC analysis of these formulations showed

4.812 mg of forskolin in 1 g of Makandi ghanavati

and 3.51 mg of forskolin in 1 g of Makandi churna.

As already stated forskolin is responsible for many

pharmacological actions related to cardio vascular

system. It was hypothesized that the forskolin present

in these two formulations may be responsible for the

anti-atherogenic and cardioprotective effects.

However, the results obtained did not correlate with

the forskolin content. Makandi churna with lower

forskolin content showed better cardioprotective

activity in comparison to Makandi ghanavati which

had higher forskolin content. This indicates that other

constituents may also play a major role. Elucidation

of the full chemical profile may throw light on the

nature of other constituents.

Acknowledgement

Authors are thankful to Prof. M.S. Baghel,

Director, IPGT & RA, Gujarat Ayurved University,

Jamnagar for the constant encouragement and for

providing facilities to carry out this study.

References 1 Bhushnam, Lala Shaligramji Vaishya Virachita Shaligram

nighantu, Khemaraj Shrikrushnadasa Prakashana, Mumbai,

Parishishta Bhaga, Sanskaran, 1993, p. 931.

2 Nighantu Ratnakara, A compendium of system of Hindu

medicine, edited by Bhishagvarya Late Krishna Shastri R.

Navre collated with spacious notes by Vasudev Laxman

Shastri Pansikar & Krishnaji Vitthal Soman, Part I,

Gunadosha Prakarana, 1993, p. 152.

3 Metzer H and Linder E, FORSKOLIN: Its chemical,

biological and medical potential, IRCH Med Sci, 1981, 9, 99.

Table-6 Effect of Makandi ghanavati and Makandi churna on cardiac output


Cardiac output

(ml/min)

20.90 ± 03.40 10.10 ± 01.40* 11.60 ± 04.90 09.70 ± 03.50* 07.60 ± 02.30 23.20 ± 08.10

LCW

(Kg.min/m2)

0.0269 ± 0.0074 0.0162 ± 0.0024 0.0115 ± 0.0052 0.0141 ± 0.0057 0.0108 ± 0.0033 0.0371 ± 0.0134

LVET (sec.) 0.160 ± 0.0202 0.161 ± 0.0163 0.202 ± 0.0116 0.198 ± 0.0359 0.161 ± 0.0201 0.211 ± 0.0197

PEP (sec.) 0.310 ± 0.0452 0.263 ± 0.0242 0.319 ± 0.0658 0.315 ± 0.0436 0.0436 ± 0.0425 0.294 ± 0.0587

Stroke index 4.366 ± 1.052 2.327 ± 0.308 2.913 ± 1.050 3.168 ± 1.072 1.954 ± 0.703 6.021 ± 1.954

The data were expressed as Mean ± SEM. Significant differences in each group vs. the control is *P< 0.05.


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