Studies a tfie therapeutic potential of t/ie f d t i 0 t L

STUDIES ON THE THERAPEUTIC POTENTIAL OF THE FORMULATRX-

1. STUDIES ON ANTIOXIDANT ACTTVITY

Introduction

Recent investigations in the field of free radicals have emphasized

their role in the pathophysiology of atherosclerosis. Reactive oxygen species

(ROS) have been demonstrated to be involved in the triggering of several diseases,

and compounds tliat can scavenge free radicals have great potential in

ameliorating these disease process."'

Lipids undergo peroxidative changes in the arterial wall, which

eventually produce endothelial injury. Modification introduced in LDL by free

radical mediated processes enhances foam cell production and these events are

known to be the early events in atherosclerosis.

Epidemiological studies based on dietary information indicated a

negative correlation between the presence of antioxidants in the plasma and the

incidence of coronary artery disease.242 Endogenous defence mechanism have

been identified which use antioxidants or free radical scavengers to neutralize

reactive oxygen species generated lipid peroxidation; however the extensive

generation of free radicals appears to overwhelm, the natural defence

mechanisms, dramatically reducing the level of endogenous antioxidants.243

Mort: recent studies suggest that the inflammatory tissue damage is

due to the liberation of reactive oxygen species (ROS) from the phagocytes

invading the inflammation sites.'"-' Several anti-inflammatory drugs have been

shown to inhibit production of free radicals by neutrophils, which migrate to

inflammatory sites causing tissue injury.245

A number of natural products such as ellagic acid, curcumin and

vitamins (A, C: and E) have gained wide momentum as they are shown to possess

antioxidant potential. Most of the plant polyphenols and flavonoids are good

antioxidants. In this study we have analysed the in vitro free radical scavenging

activity of the formulation, 'Caps HT2'.

Materials and methods

Animals Male Wistar albino rats (200-250 gm) were used for the study. The

animals were maintained in well-ventilated cages and fed with standard pellet

diet and water adlibiturn.

Antioxidant activity

Superoxide radical scavenging activity

Superoxide radical scavenging activity was determined by the NBT

reduction method 213 as described in the Materials and methods (2: 7a).

Hydroxyl radical scavenging activity

Measured by studying the competition between deoxyribose and the

extract for hydroxvl radicals generated from the Fe3+/ascorbate/EDTA/H202

system. Deoxyribose degradation was measured as TBARS (2: To).

Inhibition of lipid peroxidation

Ferrous ascorbate system induces thiobarbituric acid reactive

substances (TBARS) formation in liver homogenate in vitro, which is a measure of

lipid peroxidation (2: 7c).

Results

The data shown in Table 3: 1 explains the antioxidant potential of the

herbal formulation, 'Caps HT2'. The formulation was found to scavenge the

superoxide generated by photoreduction of riboflavin. The concentration needed

for 50% scavenging of superoxide was 55.00 pg/ml for the formulation and

6.32 pg/ml for the standard curcumin.

Degradation of deoxyribose mediated by hydroxyl radicals

generated by the Fe:'+/ascorbate/EDTA/H202 system was also found inhibited by

the formulation. The concentration needed for 50% inhibition was 610.00 pg/ml

and 8.93 pg/ml for the formulation and curcumin respectively.

The generation of lipid peroxides by Fe2+/Ascorbate in rat liver

homogenate was found inhibited by the addition of the formulation extract. The

concentration needed for 50% inhibition was 48.5 pg/ml whereas curcumin

needed was 2.7pg/ml.

The present study revealed the antioxidant activity of the

formulation, 'Caps HT2'. The initiating free radicals 02-, Hz02 and OH- were well

scavenged by the herbal formulation in vitro, which is comparable to that of the

known antioxidant curcumin.

Table 3: 1 - Effect of the formulation 011 free radical generation

The values are average of triplicate tubes.

-

ICso value (pg/ml)

Superoxide Hydroxyl radical

Formulation 55.00 610. 00

Curcumin 6.32 I 2.71 !

Lipid peroxide

48.50

8.93

Discussion

Although several mechanisms have been proposed to explain the

pathogenesis of cardiovascular diseases, most attention has focussed on a role for

reactive oxygen metabolites. The imbalance between reactive oxygen species and

antioxidants in the development of atherosclerosis has prompted the investigation

of antioxidant supplementation as possible therapy. Potential antioxidant therapy

should therefore include natural antioxidant enzymes or agents, which are capable

of scavenging free radicals.

Antioxidant agents of natural origin have attracted special interest

because they can protect human body from free radical mediated injury with no

side effects. Numerous medicinal plants and their formulations are used for

cardiovascular disorders in ethnomedical practices as well as on traditional system

of medicine in India. Thl? plant polyphenols and flavonoids have long been

recognised to possess antiinflammatory, antithrombotic, antioxidant and

anticarcinogenic activities.

The present study revealed the significant antioxidant activity of the

formulation 'Caps HT2'. Among the selected plants, Commiphora muku1,94 Allium

sativum,'37, li9-141 Semecarpus anacardium228 Hemidesmus i nd i~us?~~ Terminalia

aquna, 247 Tinospora cor~/ifolia,~62 Withania somnifera "-23, 178 and Ocimum

sanctum 24, '48-249 were already reported as antioxidant agents.

2. STUDIES ON ANTIPLATELET AGGREGATION ACTIVITY

Introduction

Platelets 'Ire important in the recognition of vascular injury,

formation of effectivc homeostatic plugs, retraction of clots and wound healing. 250

Platelets are also the tuajor constituents of thrombus and a source of powerful

vasoconstrictors that can cause vasospasm and enhance coagulation by

diminished blood flow.251 It is generally recognised that platelet aggregation is

abnormally increased in CAI) patients. 252

1ntravascular tluombosis is known to be the final catastrophic event

in atherosclerosis. It occur:; immediately after the rupture of the atheromatous

plaque. Various experimental and clinical evidences have established the role of

platelets as the central participant in the final thrombotic event. 253

The platelet aggregation inhibition therapy has gained much

acceptance in atherosclerosis as it limit the arterial thrombosis. In this study we

have examined the platelet aggregation inhibition efficacy of the herbal

formulation, 'Caps HT2'.

Methods

Platelet aggregation inhibition activity

Aggregation of platelet rich plasma with ADP as agonist was studied

and the effect of different concentrations of the formulation was assessed as

described in the chapter Materials and Methods (2: 9).

Results

Addition of ADP to a suspension of washed human platelets caused

a marked decrease in optical density at 600 nun indicating the aggregation of

platelets. The herbal formulation (50-150 pg/ml) interestingly inhibited platelet

aggregation in a concentration dependent manner (Fig3: 1).

Discussion

Platelets play an important role in the process of atherothrombosis

by adhering to the damaged regions of the endothelial surface. The activated

platelets form platelet-to-platelet bonds, binds also to leucocytes bringing them

into a complex process of plaque formation and growth.254

The antiplatelet therapy constitutes the best available tool for

ameliorating the rnechanis~ns related to atherogene~isl4~ and this herbal

formulation interestingly inhibited platelet aggregation. It has been revealed that

platelet aggregation il~duced by ADP, epinephrine, collagen, thrombin or platelet

activating factor could be inhibited by pre-treatment with onion as well as

garlic juice.'"'", '4% 255 The effect of gum guggulu on platelet adhesiveness was

already studied.""

Fig. 3: 1 - Effect of the formulation on ADP induced platelet aggregation inhibition

A - Control, B - Funnulation (5Opg/ml), C - Formulation (100pg/ml), D - Formulation (150pg/mI), E - Heparin (20pg/ml) and F - ADP (lOpM/d)

3. STUDIES ON ANTICOAGULANT ACTIVITY

Introduction

The obvious link between the coagulation system and atherosclerosis

has long been accepted in terms of thrombosis of a coronary artery, with the

resulting myocardial infarction.25h Reduced fibrinolytic activity and raised

prothrombotic activity are associated with hyperlipidaemia and are modified by

interventio~i.~~' Clinical artd pathological studies have shown that disruption or

erosion of atherosclerotic plaque is the major cause of coronary thrombosis.

Atherosclerosis and thrombosis are associated with many common

pathological features, such as deposition and aggregation of platelets;

monocyte/macrophage infiltration and dysfunctional endothelium and these

conditions are associated with increased oxidative stress. The coincidental

expression of various causative factors predominantly initiates blood coagulation

and clotting, leading to the thrombotic events. It is possible that inhibition of

platelet aggregation and thrombosis has a major role in decreasing cardiac events

in patients with preexisting cardiovascular disease^.^" Presently, we evaluated

the anticoagulant activity of t.he herbal formulation 'Caps HT2'.

Materials and methods

Animals The male New Zealand White rabbits (1.5 - 2kg) selected for the

study were caged in uniform hygienic conditions and fed with normal diet and

water ad Jibit~~m.

Anticoagulant activity by plasma recalcification method

The plasma recalcification time was assessed in rabbit plasma by the

intravenous administration of the formulation as described in the chapter

Materials and Methods (2: 8)'.

Results

Effect on plasma recalcification time

Normal plasma recalcification time noticed was 58 * 2 Sec in rabbits.

Administration of heparin delayed the time to 145 lt: 4 Sec (Table 3: 2), whereas the

herbal formulation delayed it to 139 t 5 Sec. Compared to heparin this drug is

more or less equally effective.

Discussion

Thrombosis is the main complication of atherosclerosis and there is

considerable evidence that the platelets of patients with atherothrombotic disease

are in an increased state of activation compared with those in healthy

individuals.25-I Even though it was not sure whether the clotting factors are casual

in the initiation and development of atherosclerosis, the various coagulation

factors associated with platelet aggregation and adhesion were seen elevated in

Table 3: 2 - Anticoagulant activity of the formulation

Sample Plasma recalcification time (in seconds)

Normal rabbit blood

Heparin (lmg/kg)

Formulation Pmg/ kg)

'The values are mean i SD of 6 animals/group

the case of atherosclerosis.

I11 this study we demonstrate that the formulation, 'Caps HT2' is

significantly potent in inhibiting thrombogenesis by enhancing the plasma

coagulation time. The clinical implication of this study was the effective use of

supplementations of the formulation to patients with vascular thrombotic

diseases.

It is reasonable to assume that an imbalance in the haemostatic

system secondary to increased clotting activity and impaired fibrinolytic functions

are of great importance in the development of thrombosis. The high TG and

VLDL have been shown to be associated with coagulation of proteins;

prothrombin and other factors explain the hypercoagulability noticed in

hypertriglycericiaen~i,~.

4. STUDIES ON LIPOPROTEIN LIPASE ENZYME ACTIVITY

Introduction

The> I~y~wrtr~glyceridacmia is an important risk factor in the

development of atherosclerosis and recent works on the development of

hypertriglycer~ciaen~la has focussed on the defects in the mechanism of their

removal of plasma VLDL and chvlon~icrons. Lipoprotein lipase is the rate-

limiting enzyme for the clearance of chylomicrons and VLDL from the plasma.

The enzyme is loc'ited in the arterial bed of many tissues but is probably

synthesized and released by parenchymal cells such as adipocytes and monocytes

for attachment to capillary endothellurn. Endothelial surface is thought to be the

principal site of action of enzyme.

A decrease in the activity of the lipoprotein lipase enzyme could

result in the accumulation of VLDL and chylomicrons in the circulation. The

lipoprotein lipase gene mlutations are associated with marked dyslipidaemia

leading to premature atherosclero~is.~~~ In view of this, agents that can enhance

the LPL activity are conside1:ed as hypolipidaemic and antiatherogenic.

The lipid lowering therapy has gained much acceptance in

atherosclerosis as it limits arterial thrombosis. Here we have examined the

efficacy of the herbal formulation, 'Caps HT2' in the treatment of cardiovascular

diseases by screening the lipoprotein lipase enzyme releasing activity.

Materials and methods

Animals

The New Zealand White rabbits (male) used for the study were caged

in uniform hygienic conditiorui and fed with normal diet and water adlibifurn.

Lipoprotein lipase enzyme releasing activity.

The Lipoprotein lipase enzyme releasing activity of the formulation

was determined by the method of Edward D. Korn (1959) "9 with the intravenous

administration of the drug (5 mg/kg body wt.) for a period of 15 min as described

in the Materials ancl Methods (2: 10).

Results

The drug treated animals for a period of 15 rnin showed increased

production of glycerol as an index of greater release and activity of enzyme from

the arterial intima l'he glycerol liberated in the drug treated animals was found

10.88 mg% whereas in the normal animals it was only 3.6% (Table 3: 3).

Discussion

Lipoprotein 1ipar;e has been reported in the post heparin plasma of

rabbits and has a major role in the transport and metabolism of triglycerides of

exogenous origin."" I t is the key enzyme, which regulates the disposal of lipid

fuels in the body .">" Consideri~ng the role of LPL in the metabolism of VLDL and

chylomicrons it could be assumed that the agent that enhance the liberation of

enzyme into the blooil can play a major role in hypertriglyceridaemia. The

glycerol liberated by the action of lipoprotein lipase enzyme in the treated animals

Table 3: 3 - Lipoprotein lipase releasing activity of the formulation in rabbits

- - - - -- --

Groups

Normal

Heparin

(lmg/ kg)

Glycerol liberated (mgl dl/ hr)

'I'lie values a r e mean + SD of 6 animals/group

was found to bt: 3 times greater than the normal untreated ones. This herbal

formulation showed an enhancing role on releasing and activating the enzyme,

effecting the metabolic degradation of lipids.

Thus this study emphasizes the lipid clearing and hypolipidaemic/

hypotriglyceridaerilic activities of the formulation, 'Caps HTT, unveiling its

antiatherogenic eiiicac).

5. STUDIES ON ANTI-INFLAMMATORY ACTIVITY

Introduction

Experimental and clinical studies, based on the markers of

inflammation and inflammatory mediators in plasma, as well as in tissue samples

obtained from atherosclerotic tissues, have provided ample evidence for the

presence of ongoing inflammation in atherosclerosis.261 In atherosclerosis,

inflammation may alter the integrity of the vascular endothelium and cause

exposure to thrombogenic material in the plaque with clot formation and

reduction in coronary flow.262

Currently, the most widely used anti-inflammatory drugs for chronic

diseases like arthritis, cardiovascular diseases etc., possess well known side effects

and none of them are suitable for prolonged use. Here we have analysed the anti-

inflammatory response of the formulation, 'Caps HT2'.

Materials and methods

Animals Male Wistar albino rats (200-250 gm) used for the study were

maintained in well-ventilated cages and fed with standard pellet diet (Lipton

India Ltd., Bangalore) and water adlibitu~r!.

Anti-inflammatory activity

Anti-inflammatory activity was studied using the carrageenan

induced acute pedai oedema and the formalin induced chronic pedal oedema in

rats as describcd in the Materials and Methods (2: l l a and 2: llb).

Results

Anti-inflammatory effect of the formulation against carrageenan-

induced acute irdlarnmation is shown in the table (3: 4). The formulation

significantly reduced the paw thickness (P<0.001) as compared to that of control

rats. The volume of the paw oedema was highest at 2nd hr after carrageenan

administration and is followed by a gradual decline (Fig 3: 2).

The results of formalin (Table 3: 5, Fig 3: 3) induced paw oedema

show that the formulation was also significantly (P<0.001) effective in chronic

inflammation. There was 42.7% reduction in oedema when the formulation was

administered at a dose of 300 mg/kg and 38.3% decrease by the higher dose (500

mg/ kg) administration.

The anti-inflammatory efficacy of the herbal formulation was

comparable to that of the standard drug acetyl salicylic acid (aspirin). In acute as

well as chronic inflammations, the 300 mg/kg dose administration showed greater

efficacy.

Discussion

Atherosclerosis is an inflammatory disease and does not result

simply from the accumulation of lipids. If we can selectively modify the harmful

Table 3: 4 - Anti-inflammatory activity of the formulation in rats (Carrageenan induced)

Groups Dose

(mg/kt: body wt.)

Control

Aspirin

Formulation

Formulation

Increase in paw thickness after 3 hrs

(cm)

% inhibition

The values arc mean f S'D of 6 ani~nals/group. ' p <0.001

Fig. 3: 2 - Effect of the formulation on Carrageenan induced pedal oedema in rats

Initial 0 hr t hr 2hr 3 hr Time inte wale

Table 3: 5 - Anti-inflammatory activity of the formulation in rats (Formalin induced)

Groups Increase in paw

Control .-. 0.47 + 0.04 -

Aspirin 100 0.29 k 0.03 ' 38.3

Formulation 300 0.27 i 0.02' 42.7

I Formulation 1 500 0.29 t 0.03 '

i ! 3 8 3

The values arc mlran + SL) of 6 ammals/ group, ' p < 0 001

Fig 3: 3 - Effect of the formulation on formalin induced pedal oedema in rats

0- I I

Initial day 3 day 5day 7%'

Treatment period

components of inflammation in the arteries and leave protective aspects intact, we

may create new avenues for the diagnosis and management of cardiovascular

diseases other than hyperch.olesterolaemia.261

Inflammation is a response of vascular tissue of the body to injury

involving infiltraticln of cells, production of mediators and release of hydrolytic

enzymes. One of the majaor causes of inflammation is the free radicals which

causes peroxiclation of membrane lipids, ageing, atherosclerosis, delayed wound

healing, oxygen toxicity, liver disorders, etc.206

The present study revealed the anti-inflammatory activity of the

formulation, 'Casps HT2'. The formulation also significantly inhibited the acute

inflammation induced by carrageenan as well as chronic inflammation induced by

formalin.

The higher concentrations of the formulation (500mg/kg) showed

the same anti-inflanimatory efficacy as that of the standard drug acetyl salicylic

acid, whereas the lesser concentration (300mg/kg) resulted in higher percentage

efficacy. The findings also indicate that increase in the free radical scavenging

activity of the formulation may be responsible at least in part, for the anti-

inflammatory activity effected.

Among the selected plants of the formulation, Withania

somnifera,'7', 178 Oci1nu11.1 sanctum 1" and Hemidesmus indicus 246 were already

reported as anti-inflammatory agents. In conclusion the anti-inflammatory

activity suggest the therapeutic use of the formulation, 'Caps HT2' in

cardiovascular anci ,issuciatetl diseases.

6. STUDIES ON FWPOLIPIDAEMIC ACTIVITY

Introduction

Myperlipiciaernia is the major risk factor in the initiation and

progression of atherosclerotic lesions. Evidence from studies both in animals and

humans indicate that atherosclerotic progression can be slowed if elevated serum

concentration of the atherogenic lipoprotein and triglycerides are reduced, which

in turn prevents coronary heart disease.

Chronic ingest:ion of atherogenic diet containing cholesterol was

known to produce marked hyperlipiclaemia, which is one of the major correctable

risk factors for causing coronary heart diseases. Raised levels of low-density

lipoprotein cholesterol (LDL) as well as reduced high-density lipoprotein

cholesterol (HDL) are risk factors in atherosclerosis. Lipid peroxides also

contribute to the development of atherosclerosis, possibly due to the oxidation of

LDL. High levels of lipid peroxidation products have been associated with an

atherogenic lipid pr12fiLe.2~~

The risk of cosronary heart disease (CHD) can be lowered by

treatment that reduces the plasma cholesterol concent~at ions .~~~ Presently we

examined the hypocholesterolaemic/hypolipidaemic effect of the herbal

formulation, 'Caps HT2' on serum lipid parameters in diet induced

hyperlipidaemic/hypercholesterolaemic rats for a period of 30 days.

Materials and methods

Animals Male Wistar albino rats (200-250 g) used for the study were caged in

uniform hygienic collditlons and fed with standard pellet diet and water ad

libitum.

Hypolipidaernic activity

The rats were made hyperlipidaemic by the administration of

atherogenic diet and the serum lipid parameters were estimated. The detailed

method was given in the chapter Materials and Methods (2: 13a - 2: 13e)

Results

The rats fed with HFD during the experimental period showed a

mean body weight increase of 33.3% (B), whereas the normal rats showed an

increase of only 15.1% (Table 3: 6). Simultaneous administration of the herbal

formulation during the experimental period of one month showed a highly

significant decrease in body weight compared to HFD fed group B (Fig 3: 4).

Expcri~ncntal lhyperlipidaemia in rats was associated with an

increase in serum lipid p:rofile.ZG Treatment with the herbal formulation

significantly changed the lipid parameters (Table 3: 6). The administration (300

and 400 mg/kg body wt) for a period of 30 days was associated with a significant

(P<0.001) decline in total cholesterol, LDL cholesterol, triglycerides and

phospholipids. A highly significant (P<0.001) increase in the HDL cholesterol was

also noticed by the administration of all the four different concentrations

(Table 3: 6) of the formulation.

Table 3: 6 - Effect of the formulation on serum lipid profile of rats fed with HFD for a period of 30 days.

Groups

A . \T,,,,l ' . L .".'.."' -% - HFD

Total Cholesterol Cholesterol

(mg/dl) !mg/dl) i

Cholesterol / Atherogen~c (mg/dl\ Index

-----C-- 18.4k0.9 I O.S:fG.O5 I

Triglycerides

(me!dl)

Phospholipids 1 % wt. '

( m I lilcrease

L - Lovastatin (5mg/kg), R - Formulation (100 mg/kg). F2 - Formulation (200 mg/kg), Fg- Formulation (300 mg/kg), Fa- Formulation (400 mg/kg). "B compared to A, 'C, 'D, 'E, 'F and 'G compared to B, ' P <0.001, " P <0.005, -' P <0.01

ng. 3: .. 4 .- - - . - - Peentag* ..... - ch%g+ in bdj l weight of n;t51 @~atgd Nth Em) and the f~rrnulation for P pe~iod of 30 days

A e B E P G Experimental groups

Fig 3: ti show the percentage change in lipid profile of the various

drug treated groups compared to normal rats (A). Administration of the

formulation (300 anci 400mg/kg body wt.) for one month was associated with a

significant (P<O.UO1) ~Iccline in total cholesterol (23-24%), LDL cholesterol (50-

53%), triglycerides (4044%) and phospholipids (27-28%), (Fig 3: 6). The efficacy

of the formulation was found greater than that of the standard drug lovostatin

(table 3: 6) and is also evident in relation to the atherogenic index (Fig 3: 7). The

atherogenic index was 0.91 in the case of lovostatin fed group (C) where as it was

reduced 0.71 anci 0.72 in the c:ase of the formulation treated groups (F and G). The

HDL cholesterol showed 28-36% increase (Fig 3: 6) in concentration in relation to

the MFD fed group (B) with the administration of the formulation, 'Caps HT2' and

exceeded in the increase in percentage (27%) with that of the standard .drug

lovostatin.

Discussion

Association of dietary lipids with hyperlipidaemia especially

hypercholesterolaernia anci the latter wit11 the development of atherosclerosis was

well established. Reduction in blood lipids/cholesterol especially from LDL

fraction could be used as; a means oi arresting the development of

atheros~lerosis.~66 Cholesterol feeding in experimental animals induces marked

change in the conlposition and atherogenicity of plasma lipoproteins.

In thc present study, administration of the herbal formulation,

'Caps HT2' resulted in a highly signiiicant reduction in total cholesterol, LDL

cholesterol triglyccricies and phospholipids with a concomitant rise in HDL

Fig. 3: 5 - Percentage change in the serum lipid profile of rats treated with HFD and formulation compared to normal rats

for a period of 30 days

Experimental groups .TC WHDL WLDL WTG WPL

B - HFD, C - HFD+ Lovastatin (5mg/kg), D - HFD+ Formulation (100 mg/ kg), E - HFD+Formulation (200 mg/ kg), F - HFD+Fonnulation (300 mg/kg), G - HFD+Formulation (400 mg/kg).

Fig. 3: 6 - Percentage change in the lipid profile of rats treated with HFD and formulation compared with HFD fed for a period of 30 days

-60 ~+--- I I I I

C D E F G Groups

C - HFD+ Lovastatin (5mg/kg), D - HFD+Formulation (100 mg/kg), E - HFD+ Formulation (200 mg/ kg), F - Hm)+ Formulation (300 mg/kg), G - HFD+ Formulation (400 mg/ kg)

Fig. 3: 7 - Atherogenic index of rats treated with HFD and the formulation for a period of 30 days

D Experirrrental groups

A - Normal, B - HFD, C - HFD+ Lovastatin (51ng/kg), D - HFD+Fonnulation (100 mg/kg), E - HFD+ Formulation (200 mg/kg), F - HFD+ Formulation (300 rng/kg), G - HFD+ Formulation (400 mg/kg).

cholesterol. 'I'lic pel-rentagcb decrease in body wctiglit observed betrvcen control

and drug treated groups t v a j indicative of its efficacy against obesity. Changes in

the concentrations i ~ t LDt, cholesterol and HDL cholesterol induced by therapy

retard or reverse tlic flow of cliolcsterol into atheromatous lesions, thereby

improving the stability of plaques and lowering the risk of plaque disruption.^

It is possible that the beneficial cardiovascular effects of the

formulation may be related to its antioxidant, anticoagulant, hypolipidaemic,

platelet aggregation ir~liibitic~n, and lipoprotien lipase releasing properties. It is

evident froni the: results that the 300 and 400 mg/kg body wt. of the formulation

administration exerteci significant I~ypocholesterolaemic, hypotriglyceridaemic

and hypophospholip~daen~ic effects with a highly significant rise in good

cholesterol (HDL). Thus the present study substantiates clearly the effectiveness

of the herbal formulation, 'Caps HT2' as a hypocholesterolaemic/hypolipidaemic

and antiatherogenic agent and could be used in the amelioration of cardiovascular

and related disease conditions.