Download - Doctor of Medicine in Pharmacology
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