COMPARISON OF ANALGESIC AND

ANTIINFLAMMATORY ACTIVITY OF

AQUEOUS AND ETHANOLIC EXTRACT OF

ASPARAGUS RACEMOSUS WITH ASPIRIN

DISSERTATION SUBMITTED TO

THE TAMILNADU DR.M.G.R. MEDICAL UNIVERSITY,

CHENNAI

In the partial fulfillment of regulation for the award of the Degree of

Doctor of Medicine in Pharmacology

M.D. BRANCH – VI

INSTITUTE OF PHARMACOLOGY

MADURAI MEDICAL COLLEGE

MADURAI

MARCH 2009

CERTIFICATE

This is to certify that the dissertation entitled

“COMPARISON OF ANALGESIC AND

ANTIINFLAMMATORY ACTIVITY OF AQUEOUS AND

ETHANOLIC EXTRACT OF ASPARAGUS RACEMOSUS

WITH ASPIRIN” is a bonafide record work done by

Dr.B.MAHARANI under my direct supervision and guidance

in the Institute of Pharmacology, Madurai Medical College,

Madurai during the period of his post graduate study for MD,

Branch VI –Pharmacology from 2006 – 2009.

Place: Madurai Director & Professor,

Institute of Pharmacology,

Date: Madurai Medical College

Madurai.

DECLARATION

I Dr.B.MAHARANI solemnly declare that the

dissertation titled “COMPARISON OF ANALGESIC AND

ANTIINFLAMMATORY ACTIVITY OF AQUEOUS AND

ETHANOLIC EXTRACT OF ASPARAGUS RACEMOSUS

WITH ASPIRIN” has been prepared by me under the able

guidance and supervision of my guide Dr.R.MEHER ALI.

M.D., former Director and Professor of Pharmacology, Institute

of Pharmacology, Madurai Medical College, Madurai in partial

fulfillment of the regulation for the award of MD

(Pharmacology) degree examination of The Tamilnadu Dr.

M.G.R. Medical University, Chennai to be held in March 2009.

This work has not formed the basis for the award of

any degree, or diploma or any other to one previously from any

other university.

Place: Madurai Dr. B. MAHARANI

Date:

ACKNOWLEDGEMENT

At the outset, I thank our Dean, Madurai Medical College,

Madurai for permitting me to carry out the study in the Institute of

Pharmacology, Madurai Medical College, Madurai.

I express sincere gratitude to my respective teacher and guide

Dr.R.MEHERALI, M.D., Former Director – Professor and Head,

Institute of Pharmacology for his constant encouragement and valuable

guidance at every stage of this study. I have gained much from his

immense wealth of knowledge and deep understanding of research

principles, that I could complete the study with little difficulty is a

testimony to his vast experience and qualities as a teacher and guide.

I recollect with pleasure the valuable support and encouragement

extended by Dr.M.SHANTHI, M.D., Additional Professor of

Pharmacology, MMC.

I am extremely thankful to my co-guide Dr.S.VIJAYALAKSHMI,

M.D., Associate Professor Pharmacology for her critical review,

valuable suggestions, unstinted co-operation at every stage for the

successful completion of the study with better precision.

I am extremely thankful to Dr.S.Thamilarasi, M.D., Associate

Professor in Pharmacology for her valuable suggestions for the

successful completion of this work.

I express my heartful thanks to Dr.R.Sarojini, M.D.,

Dr.K.M.S.Susila, M.D. Dr.R.Renugadevi, M.D. Dr.R.Navajothi, M.D.,

and Dr.R.Sudha, M.D., Assistant Professors and S.Selvakumar,

M.Pharm, Tutor for their genuine concern and interest in my work and

for their valuable suggestions during the course of the work.

I express my gratitude and profound thanks to K.Periyanayakam,

M.Pharm, Reader, Professor and Head, and all staff, Department of

Pharmacognosy for their help in the preparation of the extract.

I am indebted to Dr.A.Maheswaran, M.V.Sc., Veterinary Surgeon,

Central Animal House, Institute of Pharmacology, MMC for his help

throughout the study.

It is my duty to express my deep appreciation to my colleagues

Dr.V.Theivanai, Dr.K.Raadhika, Dr.R.Hema, Dr.K.Geetha,

Dr.A.Mohammed Gani, Dr.V.Ganesh, Dr.M.Malathi, Dr.S.Kannan,

Dr.M.Sheik Davooth, Dr.A.Lourdu Jafrin, Dr.S.Deepak,

Dr.S.Jeyaponmari, Dr.B.Jeyapriya, and Dr.B.Arulmohan for their

assistance and unflagging enthusiasm.

I thank Mr.Shenrayan, M.Sc., Head, Department of Statistics,

Madurai Medical College for his valuable help.

I take the pleasure in expressing my special thanks to all the

Technical Staff members in the Institute of Pharmacology and Central

Animal House, Madurai Medical College.

Finally, I thank my family members for their kind support and

encouragement throughout my study.

CONTENTS

S.No.

TOPIC

PAGE NO.

1.

INTRODUCTION

1

2.

AIM & OBJECTIVES

4

3.

REVIEW OF LITERATURE

5

4.

MATERIALS AND METHODS

39

5.

RESULTS

47

6.

DISCUSSION

51

7.

SUMMARY &CONCLUSION

54

8.

BIBLIOGRAPHY

9.

ANNEXURE

INTRODUCTION

Pain is “one of nature’s earliest sign of morbidity and

it stands preeminent among all the sensory experiences by which humans

judge the existence of diseases within themselves”. Pain remains the

primary reason for which patient seeks medical advice 1

. Acute pain

serves a biological purpose and is typically self limiting, ceasing when

healing is complete. Chronic pain is gravely disabling and distressing and

taxing to treat 2.

Inflammation is fundamentally a protective response

which helps the organism to get rid of microbes, toxins, necrotic cells and

tissues. Inflammation is a complex reaction, which consists

of vascular responses, migration and activation of leukocytes and

systemic reactions.

Chemical mediators of inflammation and pain are

histamine, serotonin, prostaglandins, leukotrienes, platelet activating

factor, nitric oxide, complements, cytokines and kinins. They excite

peripheral nociceptors and produce pain3.

The out come of inflammation may be beneficial when

invading organisms are phagocytosed and deleterious if it leads to chronic

inflammation without resolution. Inflammatory reactions include chronic

connective tissue diseases like rheumatoid arthritis and life threatening

hypersensitivity reactions to drugs and toxins4.

Opioid analgesics are used in the treatment of severe

pain of any type like traumatic, visceral, ischemic, postoperative, burn

and cancer pain5.

They act centrally through opioid receptors. Tricyclic

antidepressants, anticonvulsants and steroids are used as adjuvants in the

treatment of pain. Neuralgia is treated with Amitryptylline,

carbamazepine and gabapentin6. Non Steroidal Anti-Inflammatory Drugs

(NSAIDS) like acetaminophen, aspirin, indomethacin, meloxicam and

etoricoxib are preferred for the treatment of mild to moderate pain and

inflammation .They act peripherally by inhibiting the synthesis of

prostaglandins7. Diseases Modifying Anti Rheumatoid Drugs (DMARD)

like Methotrexate, Chloroquine and Gold are used to retard the

progression of degeneration and inflammation in resistant cases.

These drugs have adverse effects like gastric mucosal

damage, respiratory alkalosis, increase in bleeding tendency,

hypersensitivity reactions, renal disorders, hepatic damage etc8

.So a

search of drugs with good therapeutic effect and fewer side effects is

going on.

The planet earth has been blessed with vast majority

of flora and fauna .Most of these remains uninvestigated in the search for

biomolecules with specialized structures and target specificity. Plants are

potential sources of medicine .The various natural compounds present in

plants act on all systems of the body, having high therapeutic activity 9.

The Asparagus genus is considered to be of medicinal

importance because of the presence of steroidal saponins and sapogenins

in various parts of the plant .About 300 species of Asparagus are known

to occur in the world. Asparagus racemosus (shatavari) was

recommended successfully by Ayurvedic practitioners for the prevention

and treatment of gastric ulcers, galactogogue, antihepatotoxic and

immunomodulatory activites10.

Literature reveals that it also has analgesic and anti-

inflammatory activites11

.In the present study an attempt has been made to

evaluate the analgesic and anti-inflammatory activity of Asparagus

racemosus by comparing with the standard drug aspirin using albino mice

and albino rat.

AIM & OBJECTIVES

1. To evaluate and compare the analgesic effect of

Aqueous and Ethanolic extract of roots of Asparagus

racemosus with aspirin in albino mice.

2. To evaluate and compare the anti-inflammatory effect

of Aqueous and Ethanolic extract of roots of Asparagus

racemosus with aspirin in albino rats.

REVIEW OF LITERATURE

Pathophysiology of pain and inflammation

Animal models for screening Analgesic drugs

Animal models for screening

Anti-inflammatory drugs

Treatment of Pain and Inflammation

a. Opioid analgesics

b. Non- steroidal anti-inflammatory drugs

Botanical review – Asparagus racemosus

PAIN:

“Many, if not most, ailments of the body cause

pain”12. Pain is an unpleasant protective sensation and is the most

primitive of all senses. Sherrington defined pain as “physical adjunct of

an imperative protective reflex”13. In Latin, pain is called “poena” which

means penalty or punishment14

. It is produced by real or potential injury

to the body.

Characteristics of pain sensation15

:

Pain sensation has specific receptors and afferent fibres.

Pain continues as long as allogenic (pain causing) agent

persists.

Pain is a protopathic sensation

Pain has a large subjective component. Tolerance level

varies with individuals.

Pain is variously described as sharp, shooting, pricking,

burning, throbbing, and dull aching, sickening and may

be intermittent.

Pain may be associated with sweating, nausea, vomiting,

increased blood pressure, heart rate & increased

respiratory rate.

Pain receptors16

:

The receptors of pain sensation are the free nerve

endings. They are connected to A and C fibers. They are found in the

superficial layers of the skin, internal tissues such as periosteum, arterial

walls, joint surfaces, falx and tentorium in the cranial vault. Deeper

tissues are only sparsely supplied with pain nerve endings. Pain receptors

are nonadapting in nature. VR1 (Vaniloid Receptor-1), VRL-1 responds

to temperature above 43C and 50C respectively and produces pain.

Painful stimuli:

Multiple types of stimuli elicit pain. They are

chemical, mechanical, thermal and electrical stimuli. Bradykinin,

serotonin, histamine, potassium ions, acids, acetylcholine, proteolytic

enzymes, prostaglandins, substance-P and AMP are intrinsic chemical

stimuli that excite the pain receptors. Strong irritants, acids, alkalis, plant,

animal stings and venoms are extrinsic chemical stimuli that stimulate the

pain receptors. Muscle spasm and tissue ischemia also cause pain.

Dual pathways for transmission of pain signals17

:

Pain is either fast pain or slow pain.

Fast pain:

Fast pain is short, sharp and well localized. It is elicited by

mechanical, thermal and electrical type of stimuli. Impulses are carried by

small, myelinated A fibers to lamina-1 (lamina marginalis) of the dorsal

horns. They excite second order neurons of the neo spinothalamic tract

that terminate in ventral postero lateral nucleus of thalamus. Axons from

the thalamus reach the somatosensory area of cerebral cortex. Glutamate

is the probable neurotransmitter of the type A fast pain fibers.

Slow pain:

Slow pain is burning, aching and poorly localized pain

associated with tissue destruction. It is elicited by all type of stimuli.

Impulses are carried by unmyelinated slower C fibers to lamina II and

lamina III of the dorsal horn (substantia gelatinosa). They excite second

order neurons of the paleo spinothalamic fibers. 1/5th

of fibers terminate

in VPL (Ventro Postero Lateral) nucleus of thalamus, the remaining

fibers terminate in nuclei of reticular formation in brain stem or in tectum

of midbrain or in the grey matter, surrounding aqueduct of sylvius. Axons

from these neurons reach the somato sensory area of cerebral cortex.

Substance P is the probable neurotransmitter of the type C slow fibers.

Centers for pain sensation:

1. subcortical center – dorsomedial and intralaminar

nucleus of the thalamus. They perceive crude form of

pain sensation.

2. Cortical centers-

SSI (somato sensory) - present in postcentral gyrus

and areas 1, 2, 3.

SSII (somato sensory) - located inferior to postcentral

gyrus.

SSI & SSII are concerned with localization and

appreciation of finer forms of pain sensation.

Cingulate gyrus (area 24) - located in the medial side of

the frontal lobe is concerned with the emotional component of the pain

sensation.

Disorders stimulating pain18

:

1. Thalamic syndrome

2. Phantom limb

3. Angina pectoris

4. Causalgia

5. Intermittent claudication

6. Syringomyelia

7. Brown sequard syndrome

8. Tabes dorsalis

9. Spinal cord injury

10. Herpes zoster neuralgia

11. Trigeminal neuralgia or tic douloureux

Types of pain:

Nociceptive pain19

:

Pain that is associated with the discharges of

nociceptors is called nociceptive pain. It is classified as either somatic

pain or visceral pain. They are produced by stimulation of nociceptors.

Somatic pain:

It arises from skin, bone, joint and muscle or

connective tissue. It manifests as throbbing and well-localized pain.

Visceral pain:

It arises from internal organs such as the large

intestine or pancreas. It manifest as referred pain or as less well localized

pain and has crampy, spasmodic or aching character.

Referred pain:

It is defined as a pain that is perceived as coming from

an area that is remote from its actual origin (e.g.) pain in angina pectoris.

Inflammatory pain:

Inflammatory pain sets in area of injury more than a

minor injury and persists until the injury heals. Inflammatory mediators

stimulate nociceptors and produce pain.

Neuropathic pain20

:

A lesion of the peripheral or central nervous pathways

for pain produces neuropathic pain. Neuropathic pain includes peripheral

neuropathic pain (e.g.) diabetic neuropathy, herpes zoster neuralgia and

central neuropathic pain (e.g.) trigeminal neuralgia. Neuropathic pain has

an unusual burning, tingling or electric shock like quality and may be

triggered by very light touch. Hyperpathia (allodynia) are characteristics

of neuropathic pain.

Modulation of pain perception:

Pain perception is modulated at two places

I. At the peripheral nerve endings:

Endorphins and enkephalins combine with the

receptors at the peripheral nerve endings and decrease the response of the

receptor to nociceptive stimuli.

II. At the spinal cord level:

Two mechanisms that participate here are

i. Central pain suppressing mechanisms:

These descend to the spinal cord and modulate pain

perception.

ii. Peripheral mechanism:

Gate control Theory21

The psychologist Ronald melzack and the anatomist Patrick wall

gate proposed the gate control theory of pain in 1965.

This theory states that pain transmissions may be prevented by

innocuous inputs mediated by large myelinated afferent fibers, whereas

pain transmission may be enhanced by inputs that are carried over fine

afferent fibers.

INFLAMMATION:

Inflammation is a complex reaction to injurious agents

such as microbes and damaged cells usually necrotic that consist of

vascular response, migration and activation of leucocytes and systemic

reactions. The inflammatory process is closely intervened with the

process of repair. Inflammation is fundamentally a protective response.

Inflammation and repair may be potentially harmful e.g. rheumatoid

arthritis.

Inflammatory diseases cover a broad spectrum of

conditions including rheumatoid arthritis, osteoarthritis, inflammatory

bowel diseases, multiple sclerosis, asthma etc. Although inflammation is

the unifying factor, treatment approach required for each of the

inflammatory disease is often unique. Each patient population has distinct

therapeutic needs that are inadequately served by current prevention and

treatment strategies. Cytokines have been shown to play central roles in

inflammatory diseases and inhibition of their action or activation is a

proven or promising approach to modulation of these diseases. Few years

ago, inflammatory disorders were treated primarily with nonselective

anti-inflammatory drugs, however, now days specific mediator

antagonists alone or in combination and gene therapy are also being tried.

Efforts to develop new safer and more effective anti

inflammatory drugs are based on the improved understanding of the role

of key mediators identified as the key culprits in this malady.

Agents causing inflammation:

1. Physical agents –heat, cold, radiation, mechanical trauma

2. Chemical agents - organic and inorganic poison

3. Infective agents –bacteria, viruses, toxins

4. Immunological agents –cell mediated and antigen

antibody reactions

Signs of inflammation:

Celsus in 1st century A.D named four cardinal signs of

inflammation as

Rubor

Tumor

Calor

Dolor

Virchow later added 5th

sign as functio laesa

Types of inflammation22

:

Based on clinical onset and duration, inflammation may be

Acute –within minutes to hours

Sub acute –days to weeks

Chronic –weeks to months

Mediators of inflammation23

:

Inflammation is a multicellular and multi molecular process.

Mediators of inflammation are

1. Cells

Intravascular – neutrophills, lymphocytes, monocytes,

platelets, esionophills and basophills.

Extra vascular – mast cell, fibroblast and macrophage.

Vascular – endothelium.

2. Molecules:

Structural fibrous proteins –collagen, elastin.

Adhesive glycoproteins -fibronectin, laminin, tenascin,

non fibrillar collagen and others.

Proteoglycans.

3. Messangers:

Chemokines:

Cytokines meant for chemotaxis of cells are

called chemokines. They are the pathogenic mediators of

asthma, allergic diseases, infections, cancer, rheumatoid

arthritis and sarcoidosis.

Cytokines:

a. Regulating lymphocyte function – IL-2

(Interleukin), IL-4, IL-10 and TGF-(Transforming

Growth Factor)

b. Cytokines involved in natural immunity – TNF

(Tumor Necrosis Factor), IL-1 and Interferon.

c. Cytokines activating inflammatory cells – IFN-,

TNF-, IL-5, IL-10 and IL-12.

d. Cytokines stimulating haematopoiesis – IL-3, IL-7,

C-Kit ligand, CSF (Colony Stimulating Factor) for

granulocytes, macrophages and stem cell factors.

Adhesion molecules – E -selectin, P- selectin, L- selectin,

lewis- X, ICAM-1, 2 integrin and PECAM-1.

4. Plasma proteins:

Complement proteins

Immunoglobulins

Acute phase reactants

Coagulation factors

Fibrinolytic system

Acute Inflammation24:

It is the immediate and early response to injury.

Steps involved in acute inflammation:

The vascular changes in acute inflammation are

characterized by increased blood flow secondary to arteriolar and

capillary bed dilatation. Increased vascular permeability results in

exudates of protein rich extra vascular fluid. The leukocytes (neutrophils)

first adhere to the endothelium via adhesion molecules, then leave the

microvasculature and migrate to the site of injury under the influence of

chemotactic agents. Phagocytosis of the offending agent occurs, which

leads to death of infectious microorganisms. During chemotaxis and

phagocytosis, activated leukocytes releases toxic metabolites and

proteases extracellularly, causing endothelial and tissue damage (pain and

loss of function).

Outcomes of acute inflammation:

The nature and intensity of the injury, the site and tissue

affected and the ability of the host to mount a response modify the

consequences of acute inflammation. Acute inflammation has one of four

outcomes.

1. Complete resolution- occurs when the injury is limited or

short lived when there was little tissue destruction and when the tissue

was capable of regeneration.

2. Abscess formation- occurs in the setting of certain

bacterial or fungal infections.

3. Progression to chronic inflammation.

Chronic inflammation25:

Chronic inflammation is considered to be inflammation of

prolonged duration (weeks or months) in which active inflammation,

tissue destruction, and attempts at repair are proceeding simultaneously.

It is characterized by

a. Infiltration with mononuclear cells.

b. Tissue destruction induced by inflammatory cells

c. Repair involving new vessel proliferation and

fibrosis.

Chronic inflammation arises in the following conditions.

(i) Persistent infections – due to mycobacteria,

trepenoma pallidum, certain fungi.

(ii) Prolonged exposure to potentially toxic

agents. e.g. silica, elevated lipids.

(iii) Autoimmune diseases.

Types of chronic inflammation:

Conventionally it is subdivided in to 2 types.

1. Non-Specific.

2. Specific

Histologically, it is subdivided in to two types

1. Chronic non-specific inflammation- it is

characterized by nonspecific inflammatory cell

infiltration. e.g. Chronic osteomyelitis and lung

abscess.

2. Chronic granulomatous inflammation – it is

characterized by formation of granulomas e.g.

tuberculosis, leprosy, syphilis, actinomycosis,

sarcoidosis etc.

Out come of Chronic Inflammation:

Scarring or fibrosis – occurs when inflammation occurs in

tissues that do not regenerate. Extensive fibrinous exudates that cannot be

completely absorbed are organized by growth of connective tissue

elements, resulting in the formation of a mass of fibrous tissue.

Histological patterns of inflammation:

1. Serous inflammation – found in body cavities lined by

mesothelial cells. Cell-poor and fluid rich transudates are

found.

2. Fibrinous inflammation – severe than serous inflammation,

it has high vascular permeability, fibrin forms in the

inflammatory site.

3. Suppurative / Purulent inflammation – caused by

pyogenic bacteria like streptococci and staphylococci. It has

neutrophil rich exudates, marked tissue necrosis, abundant

neovascularization during the process of resolution.

Systemic effects of inflammation:

Acute phase response – it has endocrine, autonomic

and behavioural changes.

Fever- it is the commonest symptom and sign that

suggests inflammation. It is associated with myalgia, backache,

arthralgia, anorexia, somnolence, chills and rigors.

The inflammation is followed by body’s attempt to

heal the damage, the process of repair and it involves cell proliferation,

differentiation and extra cellular matrix deposition.

Animal models for screening analgesic and anti-

inflammatory drugs:

Animal models for Analgesic Drugs26, 27, 28

:

Pain is very difficult to define and to measure. The

response to noxious stimuli depends upon the instantaneous states of parts

of the nervous system and their interactions. Analgesics obtund the

responses to noxious stimuli. Animal models for screening analgesic

drugs are

1. Haffner’s tail clip method

2. Eddy’s hot plate method

3. Radiant heat method

4. Tail warm water immersion method

5. Tooth pulp electrical stimulation method

6. Monkey shock titration test

7. Measured caudal compression test

8. Pethidine potentiation

9. Nalorphine antagonism

10. Oxytocin cramping

11. Lenticular opacity

12. Formalin test

13. Writhing induced by chemicals

14. Randall selitto test

15. Neuropathic pain model

16. Pododolorimeter

17. Rectodolorimeter

Among the different available methods, Haffner’s tail

clip and Eddy’s hot plate method were utilized to evaluate analgesic

activity of test compound.

Animal models for Anti-inflammatory Drugs29

:

Present day anti inflammatory drug discovery is based

on preliminary in vitro observation in a number of standard anti-

inflammatory assays.

In vitro methods:

1. Inhibition of NO production induced by IFN- in

mouse macrophages.

2. Measurement of NO production in mouse

macrophages

3. Mast cell degranulation

4. Adhesion assays

5. Cyclooxygenase assays

The effective candidate drug in vitro tests is later

tested in whole animal models of acute, subacute and chronic

inflammation.

In vivo methods:

1. UV-B induced erythema in guinea pigs

2. Chemically induced paw edema

a. Carrageenan

b. Egg white

c. Kaolin

d. Mustard

e. Dextran

3. Pleural exudation method

4. Cotton pellet induced granuloma.

5. Freund’s adjuvant arthritis

6. Papaya latex induced arthritis

7. Air pouch model

8. Croton oil induced ear edema in mice

9. Arachidonic acid induced ear edema in mice

10. Anti-inflammatory activity against erythema caused

by radiation.

11. Tuberculin sensitivity

12. Inhibition of ascites

13. Hyaluronidase inhibition.

Among different models for anti-inflammatory

activity, carrageenan induced rat paw edema method is employed

commonly. Most of the currently available anti rheumatic drugs have

shown activity in this model, which utilize the transudative and exudative

phases of inflammation. Therefore for evaluation of anti-inflammatory

activity of a new compound we can rely on the reduction of transudative

and exudative phase of inflammation Viz carrageenan induced foot paw

edema method.

TREATMENT OF PAIN AND INFLAMMATION:

Historical events30, 31

:

In 1803 Serturner isolated the pure alkaloid morphine

from crude opium –named after Greek god of dreams Morpheus.

Morphine is the prototypical opioid agonist. Willow bark had been used

for many centuries for the treatment of fever and pain. Salicylic acid was

prepared from this plant, which is the protype Non Steroidal

Antiinflammatory Drug (NSAID).

Non-Pharmacological management of pain32

:

Transcutaneous electrical nerve stimulation, hot or cold

packs, massage and acupuncture these therapies were designed to activate

the gate control mechanism. Biofeedback, chiropractic, meditation, music

therapy, cognitive behavioral therapy guided imagery; cognitive

distraction and framing may be of help in treating pain.

Pharmacotherapy:

Analgesics are of two broad types, opioid and non-opioid drugs.

Opioid analgesics:

Mechanism of action:

They act as complete or partial agonist at opioid receptors.

There are several types of opioid receptors present in brain, spinal cord

and in the periphery e.g. gut. Three subtypes of opioid receptors involved

in analgesia are , , receptors. Most of the narcotic analgesics act

through receptors. Morphine also act on and receptors. These drugs

mimic the action of endogenous peptide neurotransmitters leu and met

enkephalin. Naloxone is an antagonist at all the receptors. Buprenorphine

and pentazocine is partial agonist at the receptor and pentazocine is an

agonist at the receptor33

.

Opioid analgesics are used in the treatment of severe pain of

any type like postoperative pain, cancer pain, neuropathic pain, angina

and also relieves anxiety associated with pain34

.

Morphine is high efficacy opioid analgesic used most

commonly for analgesic and non-analgesic purposes and it is obtained

from opium. Opium is dried juice obtained from the unripe seed capsule

of papaver somniferum35

.

Opium contains about 20 alkaloids. These alkaloids are

divided in to 2 groups36

i. Phenanthrenes

ii. Benzylisoquinolines

Phenanthrene derivatives:

Morphine

Codeine

Thebaine

Benzylisoquinolines derivatives:

Papaverine

Noscapine

Commonly used opioid analgesics

1. Natural opium alkaloids

Morphine

Codeine

2. Semisynthetic opiates

Diacetyl morphine

Pholcodeine

Hydromorphone

Oxymorphone

Hydrocodone

Oxycodone

3. Synthetic opioids

Pethidine

Fentanyl, sufentanil, alfentanil, remifentanil.

Methadone

Dextro propoxyphene

Tramadol

Mepridine

Levorphanol

Common adverse effects of opioid analgesics37

:

Nausea

Vomiting

Constipation

Respiratory depression

Histamine release

Hypotension, bradycardia

Increased intracranial pressure

Spasm of smooth muscles (bronchus, biliary tract,

urinary tract)

Tremors, delirium, urticaria, rash, anaphylactoid

reaction are rare adverse effects.

Morphine produces tolerance and dependence on

chronic administration.

Acute opioid toxicity:

It results from clinical overdosage, accidental over

dosage in addicts or attempts at suicide .The triad of coma, pinpoint

pupils , depressed respiration suggests opioid poisoning .Naloxone is the

treatment of choice .

Different routes of administration of opioid analgesics:

1. Oral

2. Parenteral

3. Patient controlled analgesia

4. Intraspinal infusions

5. Peripheral analgesia-topical

6. Rectal administration

7. Administration by inhalation

8. Oral transmucosal administration

9. Transdermal or iontophorectic application.

10. Peripheral administration e.g. knees

Newer aspects38

:

Nefopam –used for control of pain in opioid addicts

Tricyclic anti depressants – Amitryptylline ,Venlaflaxine

are used in severe painful neuropathy .

Gabapentin – an anti convulsant GABA analog, used for

the treatment of neuropathic pain.

Ketamine –NMDA antagonist improves analgesia and

reduces opioid requirements under condition of opioid

tolerance.

Future prospects:

Enkephalinase inhibitors-Thiorphan inhibits metabolic

degradation of enkephalins

VR-1 antagonists may be the useful target for controlling

peripheral pain during inflammation

Lidocaine and Mexiletine are useful in some chronic pain

states. It acts by blocking PN 3/SN 3-sodium channel in

nociceptive neurons in dorsal root ganglia.

Ziconotide –blocker of voltage gated N –type of calcium

channel. Used as intrathecal analgesic in patients with

refractory chronic pain.

Nicotine and its analogs – their use for postoperative

analgesia is under investigation.

9

- tetra hydro cannabinol –interacts with TRPV-1

capsaicin receptors to produce analgesia under certain

circumstances

Oral pregabalin, Duloxetine – used for the treatment of

painful neuropathy.

Non-steroidal anti inflammatory drugs:

The non-opioid analgesics as a group39

Relieve pain without interacting with opioid receptors

Reduce elevated body temperature (antipyretic effect)

Possess anti-inflammatory property

Have antiplatelet activity to varying degree

Do not cause sedation and sleep

Are non addicting

Mechanism of action:

These drugs act primarily on peripheral pain

mechanisms, and also in the CNS to raise the pain threshold.

Prostaglandins, prostacyclin, thromboxane A2 are produced from

arachidonic acid by the enzyme cyclooxygenase, which exists in a

constitutive (Cox-1) and an inducible (Cox-2) isoforms. Cox-1 serves

physiological house keeping functions, while Cox-2 is induced by

cytokines and it is responsible for the production of prostaglandins at the

site of inflammation. Now recently Cox-3 has been identified which is

expressed most abundantly in cerebral cortex and heart. It is involved in

pain perception and fever and not in inflammation.

NSAID’S acts by inhibiting Cox enzymes. Some

drugs also act by inhibiting adhesion molecules.

Commonly used NSAID’ S:

I. Non selective irreversible inhibitors of Cox: Aspirin,

sodium salicylate, sulfasalazine, olsalazine, methylsalicylate

II. Non selective reversible inhibitors of Cox:

Phenylbutazone, oxyphenbutazone, indomethacin, sulindac,

ibuprofen,ketoprofen, flurbiprofen, naproxen, mefenamic

acid, flufenamic acid, tenoxicam, piroxicam, ketorolac,

tolmetin, oxaprozin, diflunisal, diclofenac sodium, tiaprofen,

azapropazone and carprofen.

III. Weak inhibitor of Cox-1 and Cox-2 and other modes of

anti-inflammatory action: Nimesulide

IV. Preferential Cox-2 inhibitors: Meloxicam, etodolac,

nabumetone.

V. Selective Cox-2 inhibitors: Rofecoxib, celecoxib,

valdecoxib, etoricoxb, parecoxib, lumiracoxib.

VI. Cox-3 inhibitor (or) reversible inhibitor of cox-1:

Paracetamol, metamizol.

VII. NSAID’S which do not inhibit prostaglandin synthesis:

Nefopam

In addition to this glucocorticoids also have powerful

anti-inflammatory effects. Lipooxygenase inhibitors and Leukotriene

receptor antagonists also have anti-inflammatory activity and are used in

the treatment of asthma. The NSAID’S continue to have a significant role

in the long-term treatment of arthritis. Disease modifying anti rheumatic

drugs (DMARD’S) slows the bone damage associated with rheumatoid

arthritis and are thought to affect more basic inflammatory mechanisms.

They are:

Chloroquine

Hydroxy chloroquine

Sulfasalazine

Leflunomide

Mycophenolate mofetil

Gold

Penicillamine

Methotrexate

Chlorambucil

Cyclophosphamide

Cyclosporine

Azathioprine

TNF- Blocking agents

- adalimumbab

- Infliximab

- Etanercept

Abatacept

Rituximab

Anakinra

Common adverse effects of NSAID’S:

Gastrointestinal – gastric irritation, erosions, peptic

ulceration, gastric bleeding, perforation and oesophagitis.

Renal – Sodium, water retention and chronic renal failure.

Hepatic- Raised transaminases.

CNS – Headache, mental confusion and behavioural

disturbances

Haematological - bleeding and thrombocytopenia.

Interstitial nephritis, papillary necrosis and Reye’s syndrome

are rare adverse effects.

Asthma, angioneurotic swelling, urticaria, rhinitis and nasal

polyposis occurs in susceptible individuals.

Future prospects40

:

1. Nitric oxide plays a major role in maintaining the

integrity of gastric mucosa. Nitro aspirins – GI friendly

aspirins – helps in preventing aspirin induced GI lesions.

2. Cox-2 enzyme plays a role in bone repair. Cox-2

inhibitors can be used short term in postoperative

orthopedic cases.

3. Fully humanized anti- TNF- antibody, D2E7 was under

trial as DMARD.

4. Tenidap sodium as a DMARD – it is anti IL-1 inhibitor

and IL-1 receptor antagonist.

Surgical approaches to pain management:

1. Ablative procedures –

a. Regional nerve blockade with local anesthetics

(lidocaine) are used to manage transitory, severe,

localized pain.

b. Epidural blockade- when pain is localized to one or two

dermatomal segments.

c. Sympathetic blockade – in the treatment of causalgia and

related disorders.

2. Peripheral surgical approaches – peripheral nerve and dorsal

nerve root transection can be done in patients with truly

segmental truncal pain syndromes.

3. Central surgical approaches

- Anterolateral spinal cordotomy

- Prefrontal lobotomy

- Posterior rhyzotomy

Botanical Review

Asparagus racemosus

The history of medicine in India can be traced to the

remote past. The earliest mention of the medicinal use of plants is found

in the rig-veda. Susruta samhita written on 1000 B.C. contains a

comprehensive chapter on therapeutics. Charaka samhita gives a

remarkable description of the materia medica41

. Medicinal plants have

played a dominant role in the introduction of new therapeutic agents.

Discovery and development of new therapeutic agent is a continuing

process in India, there are over 15000 kinds of naturally occurring higher

plants. About 2000 species of these are reported to possess medicinal

values42

.

Herbs are plants or plant products that contain

chemicals that act upon body. These preparations are derived from plants

or fungi and are believed to have healing qualities. Herbal medicines may

be useful to prevent and treat diseases and ailments or to promote health

and healing43

.

The botanical name of the plant is Asparagus racemosus.

It belongs to the Family Asparagaceae, Liliaceae .The other names of this

plant are Aparagus sarmentosus, Asparagus gonoclados, Asparagus

adscendens. The common Vernacular names of this plant are Shatavari in

Sanskrit, Shakakul in Hindi, Satmuli in Bengali, Satavar in Gujarati and

Kilavari or Tannir vittan kizhangu in Tamil .There are about 300 species

of Asparagus. It is a climber found through out tropical Africa, Java,

Australia, India and Srilanka44

.

Essential constituents45

:

It has steroidal saponins, sapogenins, sarsaponins, tannins,

alkaloids, isoflavanes.

Proven activities46

:

1. Used for the treatment of ulcerative disorders of stomach

and duodenal ulcers.

2. Enhances milk output in postpartum women

3. The extract inhibits the uterine contraction

4. As an immunomodulatory agent

5. Antihepatotoxic activity

6. Antineoplastic effect

7. Bronchodilation

Medicinal uses:

Roots and leaves have medicinal properties. It

contains large amount of saccharine matter and mucilage. It is used as

“Madhura rasam, madhura vipakam, seta-veeryam” in ayurveda and

siddha. Roots are highly mucilaginous and have antidiarrhoetic,

refrigerant, diuretic, antidysentric, nutritive, tonic, demulcent,

galactogogue, aphrodisiac, antispasmodic, analgesic and anti-

inflammatory activities. It is used externally in rheumatism and called as

“Nervine tonic” and used for nervous disorders. It is also useful in

gonorrhoea.

Toxicity47

:

Asparagus racemosus had been described as absolutely

safe for long-term use, even during pregnancy and lactation. Systemic

administration even upto oral dosages of 64gm/kg had not produced any

abnormality in behaviour pattern of mice and rat.

In the present study analgesic activity of Asparagus

racemosus is evaluated by Haffner’s tail clip and Eddy’s hot plate method

using albino mice. The Anti inflammatory activity is evaluated by

Carrageenan induced foot paw edema method using albino rat.

MATERIALS AND METHODS

A Randomised controlled, prospective animal

experimental study was carried out in the Institute of Pharmacology and

Central animal house attached to Madurai Medical College, Madurai after

obtaining Institutional Animal Ethical clearance (Annexure-1) for a

period of 10 months from September 2007 to May 2008.

MATERIALS:

Animals:

24 Albino mice

24 Albino rats

Drugs and Chemicals:

1. Aspirin

2. Aqueous extract of roots of Asparagus racemosus

3. Ethanolic extract of roots of Asparagus racemosus

4. 1% Carrageenan

5. Distilled water

Equipments:

Tail clip

Hot plate

Plethysmograph

Oral feeding tube

Tuberculin syringe

Standard drug:

Acetyl salicylate tablet is powdered and mixed with distilled

water to get the drug solution of 10 mg /ml

Test-1:

Ethanolic extract of roots of Asparagus racemosus48

100 gm of shade dried root powder of Asparagus was added

with 1L of absolute alcohol and extract was obtained by soxhlet method.

Test-2:

Aqueous extract of roots of Asparagus racemosus

250 gm of shade dried root powder of Asparagus was added with

350 ml of distilled water and extract was obtained by reflux method.

Tail clip49

:

Bulldog clamp with thin rubber sleeve was used to produce

noxious stimuli by mechanical pressure (Tail compression).

Analgesiometer or Eddy’s hot plate50

:

It was designed to study the analgesic effect of drug at different

temperatures. The temperature was monitored on a galvanometer. In this

method heat (thermal stimuli) was used as source of pain.

Carrageenan:

The main sources of Carrageenan are chondrus crispus and

gigartina mamilloza .It is a sulfated polysacchride of the red sea weed.

The name was obtained from the irish coastal town of carragheen, 1%

solution was made by dissolving 1gm of carrageenan in 100 ml of normal

saline .It causes inflammation by the release of histamine, prostaglandins,

bradykinin and 5-HT and produces edema.

Plethysmograph:

It is an apparatus containing Mercury. This apparatus has a ‘U’

shaped limb and a vertical limb. The ‘U’ shaped limb is connected to the

vertical limb through a knob. The knob is adjusted to ensure adequate

height of mercury column in the ‘U’ shaped limb. The mercury

displacement due to dipping of the paw was read from the scale attached

to the mercury column.

Oral feeding tube51

:

For mice – A polythene tube of 2-3 cm long, sleeved on a long 18-20

gauge blunted hypodermic needle was used.

For rat – A 15-16 gauge blunted hypodermic needle of 7.5-10 cm length

was used. A small ball of solder was applied around the tip and a gentle

bend about 20-30 was made 2 cm proximal to the solder.

METHODOLOGY:

Analgesic activity:

The analgesic activity of ethanolic and aqueous extract of roots of

Asparagus racemosus was evaluated in mice by the following methods.

1. Mechanical method – Haffner’s tail clip method.

2. Thermal method – Eddy’s hot plate method.

Haffner’s tailclip method:

Binachi and Franceschini demonstrated this method.

24 albino mice of 20-25 gm were selected .A tail clip was

applied to the base of the mouse tail (for 30 sec) .The reaction time (first

attempt made by the animal to remove the tail clip) was observed. Those

animals not responding to the painful stimuli within 10 sec (cut off time)

were not included for the study .The responsive animals were randomly

grouped in to 4 of 6 animals each .The drugs are administered in the

following order after over night fasting.

Groups

Treatment

Control

Distilled water

Standard

Aspirin –100mg/kg

Test-1

Ethanolic extract of Asparagus racemosus – 200mg/kg

Test –2

Aqueous extract of Asparagus racemosus –200mg/kg

Reaction time was observed before and after 2 and 4 hr of drug

administration and the results were tabulated. (Table –1) (Figure-1)

Eddy’s hot plate method:

In this method analgesic activity was evaluated by using Eddy’s

hot plate. 24 albino mice of 20-25 gm were selected .The animals were

kept over a Hot plate maintained at 50 –55 c (for 30 sec). The reaction

time (first attempt made by the animal to avoid the painful stimulus like

licking of the paws or jump response) was observed. Those animals not

responding to the painful stimuli within 10 sec (cut off time) were not

included for the study. The responsive animals were randomly grouped in

to 4 of 6 animals each. The drugs were administered to the control group,

standard group, test group 1 and 2 similar to tail clip method after over

night fasting. Reaction time was observed before and after 2 and 4 hr of

drug administration and the results were tabulated (Table -2) (Figure-2).

Anti-inflammatory activity:

Anti-inflammatory activity was evaluated by carrageenan

induced rat paw edema method.

Carrageenan induced rat paw edema method:

24 Adult albino rats weighing 175 –225 gm were randomly

divided in to 4 groups of 6 animals each. The drugs are administered in

the following order after over night fasting.

Groups

Treatment

Control

Distilled water

Standard

Aspirin –300mg/kg

Test-1

Ethanolic extract of Asparagus racemosus –1600mg/kg

Test -2

Aqueous extract of Asparagus racemosus –1600mg/kg

Immediately after drug administration 0.1 ml of

1% Carrageenan was administered sub plantarly to the left hind paw. A

mark was made in the ankle joint to ensure the dipping of rat paw to the

same level in the plethysmograph every time. Paw volume was measured

before and 3 hrs after Carrageenan administration. Mean paw volume in

the control, standard, test –1 and test –2 was found .The percentage of

inhibition of edema in the treatment group was calculated by using the

following formula.

Percentage of inhibition of edema = Vc-Vt /Vc 100

Vc –Mean edema in the control group.

Vt –Mean edema in the treatment group (standard, test-1 and test-2)

The results were analyzed by using above formula and

the percentage of inhibition of edema compared. (Table -3) (Figure-3).

RESULTS

In the present study analgesic and anti-inflammatory activity

of aqueous and ethanolic extract of roots of Asparagus racemosus was

evaluated. Analgesic activity was evaluated in albino mice using tail clip

and hot plate methods. The anti-inflammatory activity was evaluated by

carrageenan induced rat paw edema method.

Analgesic activity:

Pain was induced by mechanical stimulus (Tail clip) and

thermal stimulus (Eddy’s hot plate) using albino mice. The analgesic

activity of Asparagus racemosus was evaluated and compared with

Aspirin. The results of tail clip and hot plate method were analyzed

statistically by students unpaired‘t’ test.

Tail clip method:

The mean reaction time for the control group after 2 and 4 hrs

of drug administration was 4 ± 2.61 sec and 4.5 ± 2.81 respectively,

hence it had no significant analgesic activity .The mean reaction time of

the standard group (Aspirin) after 2 and 4 hrs of drug administration was

18 ± 6.23 sec and 22.67 ± 5.72 respectively. It has statistically significant

analgesic activity (P<0.01) in comparison with control group. The mean

reaction time after 2 hrs and 4 hrs of drug administration for the test

group-1 was 24.5 ± 8.98 sec and 26.67 ± 8.17 and it had statistically

significant analgesic activity (P<0.001) in comparison with control group.

The mean reaction time for the test group-2 was 22.67 ± 9.15,

28.67 ± 3.27 after 2 hrs and 4 hrs of drug administration .It also had

statistically significant analgesic activity (P<0.001) in comparison with

control group. The results of tail clip method were tabulated (Table –1).

The results were represented graphically (Figure-1).

Hot plate method:

The mean reaction time for the control group after

2 and 4 hrs of drug administration was 4.5 ± 1.38 sec and 4 ± 1.26 sec

respectively and had no significant analgesic activity .The mean reaction

time of the standard group (Aspirin) after 2 and 4 hrs of drug

administration was 15 ± 2.19 sec and 12.5 ± 3.21 respectively. It has

statistically significant analgesic activity (P<0.01) in comparison with the

control group. The mean reaction time after 2 hrs and 4 hrs of drug

administration was 19.33 ± 8.91 sec and 18.83 ± 8.77 for test group -1

and it has statistically significant analgesic activity (P<0.001) in

comparison with control group. The mean reaction time after 2 hrs and

4 hrs of drug administration for test group-2 was 13 ± 3.16, 11.17 ± 2.14.

It also had statistically significant analgesic activity (P<0.001) in

comparison with control group. The results of hot plate method were

tabulated (Table–2). The results were represented graphically

(Figure –2).

Anti inflammatory activity:

Anti inflammatory activity was evaluated by

carrageenan induced foot paw edema method .The mean edema in the

control, standard and test group -1 & -2 were evaluated after 3 hrs of drug

administration and the percentage of inhibition of edema was calculated.

The mean paw volume in the control group was 1.08 ± 0.08 mm. There

was no significant inhibition of inflammation in the control group .The

mean paw volume in the standard group was 0.33 ± 1.005 mm .It had

69.45% of inhibition of inflammation. In the test group-1 the mean paw

volume was 0.42 ± 0.7043 mm and it had 61.11% of inhibition

of inflammation. In the test group-2 the mean paw volume was

0.33 ± 1.5905 mm and it had 69.45% of inhibition of inflammation.

The results were tabulated and compared (table-3).

The results were represented graphically (Figure-3). Hence from the

above study it was proved that Aqueous and Ethanolic extract of roots of

Asparagus racemosus had statistically significant analgesic and anti-

inflammatory activity.

DISCUSSION

Pain ‘an unpleasant sensory and emotional experience

is associated with actual or potential tissue damage’. It is the complex

group of neurophysiological process that may arise from the normal

response of the nervous system to tissue injury or pathological alterations

in neural activity or no identifiable pathology .The goal of pain

management is complete relief of pain.

Inflammation is essentially a protective response

intended to eliminate both the initial cause of cell injury and the necrotic

cells and tissues arising as a consequence of injury. Although it’s

protective it is also harmful. So treatment of inflammation is essential.

Pain and inflammation are commonly treated with

NSAID’s. Opioid analgesics are used in the treatment of severe pain. But

these drugs have common side effect like gastritis, nephrotoxicity,

hepatitis, respiratory depression, tolerance and dependence. Hence

researchers are making efforts to identify a compound with good

therapeutic effect and fewer side effects for the treatment of pain and

inflammation.

Asparagus racemosus has proven gastro protective,

galactogogue, tocolytic, anti hepatotoxic, immunomodulatory,

antineoplastic and bronchodilatory property .It has been called as

“Nervine tonic” and indicated for the treatment of nervous disorders.

The observations from the present study revealed that

the mean reaction time in Haffner’s tail clip method and Eddy’s hot plate

method are prolonged in standard and test groups compared to control.

The prolongations of mean reaction time in these groups are due to

analgesic activity of the standard drug and the extract. Statistical analysis

also revealed significant analgesic activity in the standard and test

groups-1 & 2.

In the study of Anti-inflammatory activity of the test

compound, it was found that the % of inflammation in the standard and

test groups are significantly reduced when compared with control. This

was due to anti-inflammatory activity of standard and the extract.

Asparagus racemosus has steroidal saponins,

sapogenins, sarsaponins, isoflavanes, tannins, alkaloids (asparagamine) as

essential constituents .The steroidal saponins, isoflavanes resemble

hormones. This constituent may be responsible for its anti-inflammatory

activity. The extract was safe even at the dose of 64 gm/kg. Being a

gastro protective and galactogogue the extract can be used as a safe

analgesic and anti-inflammatory agent during pregnancy. Since it has

bronchodilatory property and devoid of any hypersensitivity reactions, it

can be safely used in asthmatics as bronchodilator and anti-inflammatory

agent.

Further study directed towards the isolation and

identification of active constituents of the extract may provide an

opportunity for the development of a novel class of agent for the

treatment of pain and Inflammation.

SUMMARY AND CONCLUSION

The present study was attempted to investigate the

analgesic and anti-inflammatory property of Asparagus racemosus.

Analgesic activity was assessed by prolongation of the

reaction time in Haffner’s tail clip and Eddy’s hot plate method. The

prolongations of mean reaction time in these groups are due to analgesic

activity of the extract. Statistical analysis also revealed significant

analgesic activity in the standard and test groups-1& 2.

Anti-inflammatory effect of the extract was evaluated

using 1% Carrageenan induced rat paw edema method. The reductions in

the foot paw edema in these groups are due to anti-inflammatory activity

of the extract.Statistical analysis also revealed significant anti-

inflammatory activites in the standard and test groups 1&2.

The results of the present study indicate that the

Aqueous and Ethanolic extract of Asparagus racemosus posses significant

analgesic and anti-inflammatory effects. Hence further studies on

Asparagus racemosus may be helpful in developing a new approach for

the management of pain and inflammation with gastro, hepato and

bronchoprotective effect.

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TABLE –1

Analgesic activity of Asparagus racemosus –

Tail clip method

Groups

Reaction time (sec)

Mean ± S.D

2 hrs

4 hrs

Control

4 ± 2.60

4.5 ± 2.81

Standard

18 ± 6.23

22.67 ± 5.72

Test –1

24.5 ± 8.98

26.67 ± 17

Test -2

22.67± 9.15

28.67 ± 3.27

P<0.01

P<0.001

TABLE –2

Analgesic activity of Asparagus racemosus –

Hot plate method

Groups

Reaction time (sec)

Mean ± S.D

2 hrs

4 hrs

Control

4 .5 ± 1.38

4 ± 1.26

Standard

15 ± 2.19

12.5 ± 3.21

Test –1

19.33 ± 8.91

18.83 ± 8.77

Test -2

13 ± 3.16

11.17 ± 2.14

P<0.01

P<0.001

TABLE –3

Anti-inflammatory activity of Asparagus racemosus

Groups

Mean paw volume ± S.D

Mean % of Inflammation

Mean % of inhibition of Inflammation

Control

1.08 ± 0.08

100

0

Standard

0.33 ± 0.105

30.55

69.45

Test -1

0.42 ± 0.07

38.89

61.11

Test -2

0.33 ± 0.15

30.55

69.45

4

18

24.5

22.67

4.5

22.67

26.67

28.67

0

5

10

15

20

25

30

Mea

n r

eact

ion

tim

e(se

c)

2 hrs 4 hrs

Time

Figure-1

Analgesic activity -Tail clip method

Control

Standard

Test-1

Test-2 n=6(4)

Figure-2

Analgesic activity of Asparagus racemosus -Hot Plate method

4.54

15

12.5

19.3318.83

13

11.17

0

5

10

15

20

25

2 hrs 4 hrs

Time

Mea

n r

eact

ion

tim

e(se

c) Control

Standard

Test-1

Test-2n=6 (4)

100

30.55

38.89

30.55

69.45

61.11

69.45

0

10

20

30

40

50

60

70

80

90

100

% of

Inflammation

% of inhibition

of Inflammation

Figure-3

Antiinflammatory activity of Asparagus racemosus

Control

standard

Test-1

Test-2 n=6(4)

MEASUREMENT OF CARRAGEENAN INDUCED PAW

EDEMA – BY PLETHYSMOGRAPH

EDDY’S HOT PLATE METHOD

HAFFNER’S TAIL CLIP METHOD

MODULATION OF ENDOGENOUS OPIOID PATHWAY

BY OPIOIDS

AFFERENT PAIN PATHWAY

EFFERENT PAIN PATHWAY

MEDIATORS OF INFLAMMATION AND SITES OF

DRUG ACTION

ASPARAGUS RACEMOSUS

ROOTS OF ASPARAGUS RACEMOSUS