5. NEUROPHARMACOLOGICAL PROFILE (IRWIN S METHOD)shodhganga.inflibnet.ac.in/bitstream/10603/19320/13/13... · 2018. 7. 9. · Effects of the test substance were scored on a scale of

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5. NEUROPHARMACOLOGICAL PROFILE

(IRWIN’S METHOD)

5.1 INTRODUCTION

Neuropharmacological Profile study suggested by Irwin[228] provides

clues for the classification of the active compounds, to proceed for further

testing.[227] In this procedure mice after administering the drugs are

observed for an initial undisturbed phase followed by phases in which

they are subjected to mild to increasing stimuli.

Broadly, the neuropharmacological profile is divided into

behavioral, neurological and autonomic changes after administration of

increasing doses of drugs. Behavioral Profile includes Awareness

(alertness, visual placing, passivity and stereotypy), Mood (grooming,

vocalization, restlessness, irritability/aggressiveness and fearfulness) and

Motor activity (reactivity, spontaneous activity, touch response and pain

response). Neurological Profile includes CNS excitation (startle

response, straub tail, tremors, twitches and convulsions), Posture (Body

posture, limb posture), Motor incoordination (staggering gait, abnormal

gait and righting reflex), Muscle tone (limb tone, grip strength, body sag,

body tone, abnormal tone) and Reflexes (pinna, corneal and ipsilateral

flexor). Autonomic Profile (writhing, pupil size, palpebral opening,

exophthalmoses, urination, salivation, piloerection, hypothermia, skin

color and heart rate). Other activities observed include Respiratory rate,

Lacrimation and Death.

106

For the assessment and quantification of the above parameters,

methods described by Turner were adopted.[227] The effects of the test

substances on the animal were scored on a scale of 0 to 8. The basic

score for normal signs or effects was given 4; scores below 4 were for

subnormal responses and those above for supernormal. The base score

for abnormal signs was given 0 and the maximal score 8.

5.1.1 Behavioral Profile

1. Awareness

i) Alertness or Stupor (4): It indicates either stimulation or depression of

central nervous system.

ii) Visual Placing (4): Measures the motor co-ordination by placing animal

in different positions and noting its ability to orient itself without bump

or fall. Subnormal activity indicates motor in-coordination.

iii) Passivity (0): Measures the animal response to unaccustomed position.

It indicates tranquillization (antipsychotic), central depression, muscle

relaxant property, paralysis or anesthesia. The mouse was grasped with

the thumb and index finger, which hold the dorsal skin of the neck, while

the mouse was in a walking position. The score was given as follows: -

─ If the mouse moved its head and limbs in trying to escape 0.

─ If the mouse still grasped in the same manner, held in vertical

position and it struggles 2.

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─ If the mouse is placed in supine position on the back of the

observer’s hand held in a fist so that the thumb can support the

mouse’s head, it tries to escape 4.

─ The mouse tries to escape when held vertically by one forepaw 6 or

by one hind paw 8.

─ Intermediate scores were given when the struggle was diminished

but not abolished.

Figure 5.1 Passivity Test

iv) Stereotypy (0): is a frequent mechanical repetition of movements in

mice and was identified by searching movements of head, self-biting,

circling, tail lashing etc; it usually indicates central stimulation or

depression.

108 2. Mood

i) Grooming (4): Increased grooming indicates central stimulation or

sympathetic stimulation.

ii) Vocalization (0): Vocalization is utterance of sound and indicates to a

noxious stimulus. Generally mouse utters no sound.

iii) Restlessness (0): Indicates central stimulation and is seen as

discomfort, visceral changes or the approach of convulsions.

iv) Irritability (0): is an extension of restlessness and in the higher degrees

is demonstrated by aggressiveness.

iv) Fearfulness (0): is a normal emotional response in contrast to anxiety

and phobia to a recognized external source of danger, which is

manifested by alarm apprehension or disquiet.

3. Motor Activity

i) Spontaneous Activity (4): Spontaneous activity and reactivity measure

the stimulation of CNS or its sedation; the stimulation of ganglia and

neuromuscular junctions. When the animal is placed in a bell jar, it

searches around. Normal mouse usually shows moderate degree of

inquisitive behavior. The scores were recorded as - less activity as 4,3 or

2; little motion as 1; animal sleeps as 0.

ii) Reactivity (4): Animal was placed on the top of table after removing

from the bell jar and similar scoring as above was followed.

109 iii) Touch Response (4): indicates the presence of anesthetic activity and

involves touching the animal with pencil or forceps at various parts like

side of neck, abdomen and groin.

iv) Pain Response (4): Measures analgesia, sedation and central

depression. This method involved, attaching an arterial cramp to the

base of tail approximately 2cm from the body.

5.1.2 Neurological Profile

1. Central Excitation: Higher scores indicate CNS depressant action.

i) Startle Response (0): To startle with surprise or sudden alarm in

response to loud sound.

ii) Straub Tail (0): The degree of elevation of tail was recorded. The scores

were recorded as follows: 45° - 1; 45-90° - 1.5; 90° - 2;

iii) Tremors (0), iv) Twitches (0), v) Convulsions (0): indicate CNS

stimulation or hypoglycemia.

2. Posture

i) Body Posture (4), ii) Limb Position (4) – deviation from normal indicates

neuromuscular blockade or central disturbances.

3. Motor Incoordination

i) Staggering Gait (0): indicates ataxia.

ii) Abnormal Gait (O): indicates muscular relaxation or relates to ataxia.

iii) Righting Reflex/Somersault Test (0): Animal is tossed five times, so that

animal takes 2-3 turns in air before falling down in each trial. The scores

110 are given as: Standing on four feet in all five trials – 0; Lying on side –

1/5 or 2/5 – 2, 3/5 or 4/5 - 3; 5/5 – 3. Lying on the back –1/5 or 2/5 –

4; 3/5 or 4/5 -5; 5/5 – 6; slowly regaining normal position from a supine

or side position -7; remains in supine position – 8.

Figure 5.2 Righting Reflex

High scores indicate the presence of a depressant of CNS; an

ataractic (antipsychotic) agent; a muscle relaxant; an anesthetic; or an

agent causing synaptic blockade in some part of the nervous system.

4. Muscle tone

The tests in this group indicate muscle relaxant activity, neuronal

blockade or central depression.

i) Limb Tone (4): Grasping a forepaw of the mouse and noting the

resistance to the extension of the paw can estimate it.

ii) Grip Strength (4): Making the animal to grasp a pencil and hold in

horizontal position and noting how well it holds onto the object tests grip

strength.

111 iii) Body Sag (4), iv) Body Tone (4) and v) Abdominal Tone: Estimated by

noting the muscle tension in comparison to the control.

5. Reflexes

Decreases in the scores indicate blockade of sensory nerve, the

spinal synapses or efferent pathway.

i) Pinna reflexes (4): Observed by touching at the center of the pinna with

fine hair. The unaffected mouse draws from irritating hair.

ii) Corneal reflexes (4): By touching stiff hair to the cornea of the eye.

iii) Ipsilateral flexor (4): Toe pad is compressed with forceps, which causes

animal to flex its legs in a retiring movement.

5.1.3 AUTONOMIC PROFILE

1. Optical Signs

i) Pupil Size (4): The size of the pupil is measured and it is compared with

and without drug. Enlarged size of pupil indicates parasympatholytic

activity or sympathomimetic activity. Low score indicates muscarinic

activity.

ii) Palpebral Opening (4): It can be either widened or may become narrow.

If widened, it indicates sympathomimetic activity and if narrowed, it

indicates ataractic (antipsychotic) or sedative activity.

iii) Exophthalmoses (0): indicates sympathomimetic activity

2. Secretory signs

i) Urination (0): Indicates muscarinic activity or irritation of urinary tract.

ii) Salivation (0): Indicates muscarinic activity.

112 3. General signs

i) Writhing (0): Indicates irritation of the tissue or stimulation of sensory

receptors.

ii) Piloerection (0): Indicates a compensation for a lower body temperature

or sympathomimetic activity.

iii) Hypothermia (0): Indicates possible ataractic (antipsychotic) or akinetic

activity.

iv) Skin Color (4): Skin color especially that of the back of the ear may

change from pink to red or white. Red color indicates vasodilatation and

possible sympatholytic activity. White color may indicate vasoconstriction

or sympathomimetic activity.

v) Respiratory Rate (4): May be elevated by toxic substances or respiratory

analeptics. Respiratory depressants and agents causing central

depression, especially in higher doses, decelerate it.

v) Heart Rate (4): It may indicate parasympatholytic activity if high and

cardiotonic activity if low but there are many other possibilities.

4. Death: indicates toxicity.

5.2 MATERIALS AND METHODS

Animals:

Swiss albino mice were procured from M/s Sainath Animals

(registered with CPCSEA) one week before the experiment for

acclimatization. Mice of either sex weighing 20-25g were housed in

groups of six under standard lab conditions (temperature 25o±1oC,

113 relative humidity 55±5%, 12h:12h dark:light cycle) with standard pellet

food and water ad libitum. The overnight fasted animals (water ad

libitum) were transferred to the laboratory at least 1h before the start of

the experiment. The experiments were performed in sets of three at a

time, during the day (09:00-16:00h) and as per the guidelines of the

Committee for the Purpose of Supervision and Control of Experiments on

Animals (CPCSEA), Government of India. The Institutional Animal Ethics

Committee approved the study protocol for all the drugs

(IAEC/SUCP/01/2007, IAEC/SUCP/01/2009, IAEC/SUCP/09/2009).

Drugs:

All the drugs, Hab-e-Jund (HJ), Cynodon dactylon (CD),

Kushmanda Lehyam (KL), Itrifal Kishneezi (IK) and Barleria cristata (BC)

were administered orally in 1% gum acacia solution except Panchagavya

Ghrutham (PG), which was administered in sesame oil. Food was given

after 4h but water was given ad libitum.

Dose:

From acute toxicity studies the drugs were found to be relatively

safe and hence a dose progression of 30,100,300,500 mg/kg body weight

was followed.[227] The following doses were administered orally:

PG - 100, 300, 500 mg/kg HJ - 100, 300, 500 mg/kg

CD – 30, 100, 300 mg/kg KL - 100, 300, 500 mg/kg

IK - 100, 300, 500 mg/kg BC - 30, 100, 300 mg/kg

The mice were observed for 4h after oral administration for various

behavioral, neurological and autonomic changes as mentioned above.

114 Effects of the test substance were scored on a scale of 0-8. Base score

was 4 for the normal signs or effects. Score below 4 indicate subnormal

response and those above are supernormal. The base score for abnormal

sign was 0 and maximal score was 8.

5.3 RESULTS

PG, HJ, KL and IK showed negligible actions at 30 mg/kg body

weight. Neurological profile of these drugs at doses of 300 and 500

mg/kg body weight are shown in the Tables 5.1-5.6. For CD and BC the

activities at 100 and 300mg/kg body weight are tabulated. At higher

doses the drugs showed certain behavioral, neuronal and other changes

suggesting the course to be taken for further studies. The changes in

Neuropharmacological profile of individual drugs are mentioned below.

5.3.1 Panchagavya Ghrutham

Behavioral Profile: PG decreased alertness, spontaneous activity,

reactivity touch response and pain response indicating that the drug is a

CNS depressant and analgesic.

Neurological Profile: PG showed no CNS excitation or muscle relaxation,

but decline in reflexes were observed indicating blockade of sensory

nerves, synaptic blockade or efferent pathway.

Autonomic Profile: No gross changes were observed.

Other: Excessive defecation occurred.

Death: No death occurred.

115 5.3.2 Hab-e-Jund

Behavioral Profile: HJ decreased alertness, spontaneous activity,

reactivity, touch response and pain response (comparatively less than

PG) indicating that the drug is a CNS depressant and comparatively a

mild analgesic.

Neurological Profile: HJ showed no CNS excitation but grip strength

reduced indicting muscle relaxation. Decline in reflexes was observed

indicating blockade of sensory nerves, synaptic blockade or efferent

pathway.



5.3.3 Cynodon dactylon

Behavioral Profile: CD decreased alertness, spontaneous activity,

reactivity touch response and pain response indicating that the drug is a

CNS depressant and analgesic. The pain response was completely

abolished at a dose of 100 mg/kg body weight.

Neurological Profile: CD showed tremors, twitches and convulsions

indicating hypoglycemic activity. It also produced muscle relaxation at

300 mg/kg body weight. Decline in reflexes was observed indicating

blockade of sensory nerves, synaptic blockade or efferent pathway.


Death: One delayed death occurred at a dose of 300 mg/kg body weight.

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5.3.4 Kushmanda Lehyam

Behavioral Profile: KL at low doses increased alertness, stereotypy,

restlessness, irritability/aggressiveness, indicating that the drug is a CNS

stimulant but reversed the same at higher doses (500 mg/kg body

weight).

Neurological Profile: KL again showed mild tremors indicating CNS

excitation at low doses and completely reversed the same at higher doses.



5.3.5 Itrifal Kishneezi

Behavioral Profile: IK increased alertness, stereotypy, restlessness,

irritability/aggressiveness, indicating that the drug is a CNS stimulant.

Neurological Profile: IK showed mild tremors, twitches and convulsions

indicating CNS excitation or hypoglycemic activity. Pain response was

completely abolished at the dose of 300mg/kg body weight.


Other: Excessive defecation occurred.


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5.3.6 Barleria cristata

Behavioral Profile: BC increased alertness, stereotypy, restlessness,

irritability/aggressiveness, indicating that the drug is a CNS stimulant.

Neurological Profile: BC showed mild tremors indicating CNS excitation.

Pain response was completely abolished at the dose of 300 mg/kg body

weight.



118 Table 5.1 Neuropharmacological Profile of Panchagavya Ghrutham

300 mg/kg 500 mg/kg

Time (h) BS 0 1 2 3 4 0 1 2 3 4

Behavioral

Awareness

Alertness 4 4 4 4 3 3 4 4 3 3 2

Visual Placing 4 4 4 4 4 4 4 4 4 4 4

Passivity 0 0 0 0 0 0 0 0 0 0 0

Stereotypy 0 0 0 0 0 0 0 0 0 0 0

Mood

Grooming 4 4 4 4 4 4 4 4 4 4 4

Vocalization 0 0 0 0 0 0 0 0 0 0 0

Restlessness 0 0 0 0 0 0 0 0 0 0 0

Irritability/(Aggression) 0 0 0 0 0 0 0 0 0 0 0

Fearfulness 0 0 0 0 0 0 0 0 0 0 0

Motor activity

Spontaneous Activity 4 4 4 3 3 2 4 3 2 2 2

Reactivity 4 4 4 3 2.3 2 4 2 2 2 2

Touch Response 4 4 4 2 0 0 4 1.3 1.3 0 0

Pain Response 4 4 3 2 0 1 4 2 0 0 0

Neurological

CNS excitation

Startle Response 0 0 0 0 0 0 0 0 0 0 0

Straub Tail 0 0 0 0 0 0 0 0 0 0 0

Tremors 0 0 0 0 0 0 0 0 0 0 0

Twitches 0 0 0 0 0 0 0 0 0 0 0

Convulsions 0 0 0 0 0 0 0 0 0 0 0

Posture Body Posture 4 4 4 4 4 4 4 4 4 4 4

Limb Position 4 4 4 4 4 4 4 4 4 4 4

Motor Inco-ordination

Staggering Gait 0 0 0 0 0 0 0 0 0 0 0

Abnormal Gait 0 0 0 0 0 0 0 0 0 0 0

Righting Reflex 0 0 0 0 0 0 0 0 0 0 0

Muscle tone Limb Tone 4 4 4 4 4 4 4 4 4 4 4

Grip Strength 4 4 4 4 4 4 4 4 4 4 4

Body Sag 0 0 0 0 0 0 0 0 0 0 0

Body tone 4 4 4 4 4 4 4 4 4 4 4

Abnormal tone 4 4 4 4 4 4 4 4 4 4 4

Reflexes

Pinna 4 4 3 2.3 2 2 4 2 2 1 0

Corneal 4 4 3 2 2 2 4 2 2 1 1

I.F.R 4 4 4 4 4 3 4 4 4 2 2

Autonomic Writhing 0 0 0 0 0 0 0 0 0 0 0

Pupil Size 4 4 4 4 4 4 4 4 4 4 4

Palpebral Opening 4 4 4 4 4 4 4 4 4 4 4

Exophthalmoses 0 0 0 0 0 0 0 0 0 0 0

Urination 0 0 0 0 0 0 0 0 0 0 0

Salivation 0 0 0 0 0 0 0 0 0 0 0

Piloerection 0 0 0 0 0 0 0 0 0 0 0

Hypothermia 0 0 0 0 0 0 0 0 0 0 0

Skin color 4 4 4 4 4 4 4 4 4 4 4

Heart rate 4 4 4 4 4 4 4 4 4 4 4

Misc. Respiratory rate 4 4 4 4 4 4 4 4 4 4 4

Lacrimation 0 0 0 0 0 0 0 0 0 0 0

Dead

Number acute 0 0 0 0 0 0 0 0 0 0 0

Number delayed 0 0 0 0 0 0 0 0 0 0 0

BS-Basal Score I.F.R-Ipsilateral Flexor Reflex

119 Table 5.2 Neuropharmacological Profile of Hab-e-Jund

300 mg/kg 500 mg/kg

Time (h) BS 0 1 2 3 4 0 1 2 3 4

Behavioral

Awareness

Alertness 4 4 4 4 3 3 4 4 3 2 2

Visual Placing 4 4 4 4 3 2.3 4 4 3 3 2

Passivity 0 0 0 0 0 0 0 0 0 0 0

Stereotypy 0 0 0 0 0 0 0 0 0 0 0

Mood

Grooming 4 4 4 4 4 4 4 4 4 4 4

Vocalization 0 0 0 0 0 0 0 0 0 0 0

Restlessness 0 0 0 0 0 0 0 0 0 0 0


Fearfulness 0 0 0 0 0 0 0 0 0 0 0

Motor activity


Reactivity 4 4 4 4 3 2 4 4 2 2 2

Touch Response 4 4 4 4 3 3 4 4 4 2 2

Pain Response 4 4 4 4 3 2 4 4 4 2 2

Neurological

CNS excitation


Straub Tail 0 0 0 0 0 0 0 0 0 0 0

Tremors 0 0 0 0 0 0 0 0 0 0 0

Twitches 0 0 0 0 0 0 0 0 0 0 0

Convulsions 0 0 0 0 0 0 0 0 0 0 0


Limb Position 4 4 4 4 4 4 4 4 4 4 4


Staggering Gait 0 0 0 0 0 0 0 0 0 0 0

Abnormal Gait 0 0 0 0 0 0 0 0 0 0 0

Righting Reflex 0 0 0 0 0 0 0 0 0 0 0


Grip Strength 4 4 4 4 3 4 4 4 3 2 2

Body Sag 0 0 0 0 0 0 0 0 0 0 0

Body tone 4 4 4 4 4 4 4 4 4 4 4

Abnormal tone 4 4 4 4 4 4 4 4 4 4 4

Reflexes

Pinna 4 4 4 4 3 3 4 4 4 2 2

Corneal 4 4 4 4 3 3 4 4 4 2 2

I.F.R 4 4 4 4 4 4 4 4 4 4 4


Pupil Size 4 4 4 4 4 4 4 4 4 4 4


Exophthalmoses 0 0 0 0 0 0 0 0 0 0 0

Urination 0 0 0 0 0 0 0 0 0 0 0

Salivation 0 0 0 0 0 0 0 0 0 0 0

Piloerection 0 0 0 0 0 0 0 0 0 0 0

Hypothermia 0 0 0 0 0 0 0 0 0 0 0

Skin color 4 4 4 4 4 4 4 4 4 4 4

Heart rate 4 4 4 4 4 4 4 4 4 4 4


Lacrimation 0 0 0 0 0 0 0 0 0 0 0

Dead

Number acute 0 0 0 0 0 0 0 0 0 0 0

Number delayed 0 0 0 0 0 0 0 0 0 0 0


120 Table 5.3 Neuropharmacological Profile of Cynodon dactylon

100 mg/kg 300 mg/kg

Time (h) BS 0 1 2 3 4 0 1 2 3 4

Behavioral

Awareness

Alertness 4 4 4 4 3 3 4 3 2 2 2

Visual Placing 4 4 4 4 4 2 4 4 2 2 2

Passivity 0 0 0 0 0 0 0 0 0 0 0

Stereotypy 0 0 0 0 0 0 0 0 0 0 0

Mood

Grooming 4 4 4 4 4 4 4 4 4 4 4

Vocalization 0 0 0 0 0 0 0 0 0 0 0

Restlessness 0 0 0 0 0 0 0 0 0 0 0


Fearfulness 0 0 0 0 0 0 0 0 0 0 0

Motor activity


Reactivity 4 4 4 3 3 2 4 3 3 2 2

Touch Response 4 4 4 2 0 0 4 0 0 0 0

Pain Response 4 4 3 2 0 0 4 2 0 0 0

Neurological

CNS excitation


Straub Tail 0 0 0 0 0 0 0 0 0 0 0

Tremors 0 0 0 0 1 1 0 0 2 2 2

Twitches 0 0 0 0 0 0 0 0 0 0 0

Convulsions 0 0 0 0 0 0 0 0 0 2 2


Limb Position 4 4 4 4 4 4 4 4 4 4 4


Staggering Gait 0 0 0 0 0 0 0 0 0 0 0

Abnormal Gait 0 0 0 0 0 0 0 0 0 0 0

Righting Reflex 0 0 0 0 1 1 0 0 2 2 2


Grip Strength 4 4 4 4 3 3 4 4 3 2 2

Body Sag 0 0 0 0 0 0 0 0 0 0 0

Body tone 4 4 4 4 4 4 4 4 4 4 4

Abnormal tone 4 4 4 4 4 4 4 4 4 4 4

Reflexes

Pinna 4 4 4 4 3 3 4 4 4 2 2

Corneal 4 4 4 4 3 2 4 4 2 2 2

I.F.R 4 4 4 4 2 2 4 4 0 0 0


Pupil Size 4 4 4 4 4 4 4 4 4 4 4


Exophthalmoses 0 0 0 0 0 0 0 0 0 0 0

Urination 0 0 0 0 0 0 0 0 0 0 0

Salivation 0 0 0 0 0 0 0 0 0 0 0

Piloerection 0 0 0 0 0 0 0 0 0 0 0

Hypothermia 0 0 0 0 0 0 0 0 0 0 0

Skin color 4 4 4 4 4 4 4 4 4 4 4

Heart rate 4 4 4 4 4 4 4 4 4 4 4


Lacrimation 0 0 0 0 0 0 0 0 0 0 0

Dead

Number acute 0 0 0 0 0 0 0 0 0 0 0

Number delayed 0 0 0 0 0 0 0 0 0 0 1


121 Table 5.4 Neuropharmacological Profile of Kushmanda Lehyam

300 mg/kg 500 mg/kg

Time (h) BS 0 1 2 3 4 0 1 2 3 4

Behavioral

Awareness

Alertness 4 4 4 4 5 5 4 4 3 3 3

Visual Placing 4 4 4 4 4 4 4 4 4 4 4

Passivity 0 0 0 0 0 0 0 0 0 0 0

Stereotypy 0 0 0 0 1 1 0 0 0 0 0

Mood

Grooming 4 4 4 4 4 4 4 4 4 4 4

Vocalization 0 0 0 0 0 0 0 0 0 0 0

Restlessness 0 0 0 0 1 1 0 0 0 0 0


Fearfulness 0 0 0 0 0 0 0 0 0 0 0

Motor activity


Reactivity 4 4 4 4 4 4 4 4 2 2 2

Touch Response 4 4 4 4 4 4 4 4 2 2 2

Pain Response 4 4 4 4 4 4 4 4 2 2 2

Neurological

CNS excitation


Straub Tail 0 0 0 0 0 0 0 0 0 0 0

Tremors 0 0 0 0 1 1 0 0 0 0 0

Twitches 0 0 0 0 0 0 0 0 0 0 0

Convulsions 0 0 0 0 0 0 0 0 0 0 0


Limb Position 4 4 4 4 4 4 4 4 4 4 4


Staggering Gait 0 0 0 0 0 0 0 0 0 0 0

Abnormal Gait 0 0 0 0 0 0 0 0 0 0 0

Righting Reflex 0 0 0 0 0 0 0 0 0 0 0


Grip Strength 4 4 4 4 4 4 4 4 4 4 4

Body Sag 0 0 0 0 0 0 0 0 0 0 0

Body tone 4 4 4 4 4 4 4 4 4 4 4

Abnormal tone 4 4 4 4 4 4 4 4 4 4 4

Reflexes

Pinna 4 4 4 4 4 4 4 4 4 4 4

Corneal 4 4 4 4 4 4 4 4 4 4 4

I.F.R 4 4 4 4 4 4 4 4 4 4 4


Pupil Size 4 4 4 4 4 4 4 4 4 4 4


Exophthalmoses 0 0 0 0 0 0 0 0 0 0 0

Urination 0 0 0 0 0 0 0 0 0 0 0

Salivation 0 0 0 0 0 0 0 0 0 0 0

Piloerection 0 0 0 0 0 0 0 0 0 0 0

Hypothermia 0 0 0 0 0 0 0 0 0 0 0

Skin color 4 4 4 4 4 4 4 4 4 4 4

Heart rate 4 4 4 4 4 4 4 4 4 4 4


Lacrimation 0 0 0 0 0 0 0 0 0 0 0

Dead

Number acute 0 0 0 0 0 0 0 0 0 0 0

Number delayed 0 0 0 0 0 0 0 0 0 0 0


122 Table 5.5 Neuropharmacological Profile of Itrifal Kishneezi

300 mg/kg 500 mg/kg

Time (h) BS 0 1 2 3 4 0 1 2 3 4

Behavioral

Awareness

Alertness 4 4 4 4 5 5 4 4 5 6 6

Visual Placing 4 4 4 4 4 4 4 4 4 4 4

Passivity 0 0 0 0 0 0 0 0 0 0 0

Stereotypy 0 0 0 0 0 0 0 0 0 1 1

Mood

Grooming 4 4 4 4 5 5 4 4 4 6 5

Vocalization 0 0 0 0 0 0 0 0 0 0 0

Restlessness 0 0 0 0 1 2 0 0 1 2 2


Fearfulness 0 0 0 0 0 0 0 0 0 0 0

Motor activity


Reactivity 4 4 4 5 6 5 4 5 6 6 6

Touch Response 4 4 4 2 2 2 4 2 0 0 0

Pain Response 4 4 4 1 1 0 4 1 0 0 0

Neurological

CNS excitation


Straub Tail 0 0 0 0 0 0 0 0 0 0 0

Tremors 0 0 0 0 0 0 0 0 0 1 1

Twitches 0 0 0 0 0 0 0 0 0 0 0

Convulsions 0 0 0 0 0 0 0 0 0 0 0


Limb Position 4 4 4 4 4 4 4 4 4 4 4


Staggering Gait 0 0 0 0 0 0 0 0 0 0 0

Abnormal Gait 0 0 0 0 0 0 0 0 0 0 0

Righting Reflex 0 0 0 0 0 0 0 0 0 0 0


Grip Strength 4 4 4 4 4 4 4 4 4 4 4

Body Sag 0 0 0 0 0 0 0 0 0 0 0

Body tone 4 4 4 4 4 4 4 4 4 4 4

Abnormal tone 4 4 4 4 4 4 4 4 4 4 4

Reflexes

Pinna 4 4 4 4 4 4 4 4 4 4 4

Corneal 4 4 4 4 4 4 4 4 4 4 4

I.F.R 4 4 4 4 1 1 4 4 2 0 0


Pupil Size 4 4 4 4 4 4 4 4 4 4 4


Exophthalmoses 0 0 0 0 0 0 0 0 0 0 0

Urination 0 0 0 0 0 0 0 0 0 0 0

Salivation 0 0 0 0 0 0 0 0 0 0 0

Piloerection 0 0 0 0 0 0 0 0 0 0 0

Hypothermia 0 0 0 0 0 0 0 0 0 0 0

Skin color 4 4 4 4 4 4 4 4 4 4 4

Heart rate 4 4 4 4 4 4 4 4 4 4 4


Lacrimation 0 0 0 0 0 0 0 0 0 0 0

Dead

Number acute 0 0 0 0 0 0 0 0 0 0 0

Number delayed 0 0 0 0 0 0 0 0 0 0 0


123 Table 5.6 Neuropharmacological Profile of Barleria cristata

300 mg/kg 500 mg/kg

Time (h) BS 0 1 2 3 4 0 1 2 3 4

Behavioral

Awareness

Alertness 4 4 4 4 4 4 4 4 4 5 5

Visual Placing 4 4 4 4 4 4 4 4 4 4 4

Passivity 0 0 0 0 0 0 0 0 0 0 0

Stereotypy 0 0 0 0 1 1 0 0 0 2 2

Mood

Grooming 4 4 4 4 4 4 4 4 4 5 5

Vocalization 0 0 0 0 0 0 0 0 0 0 0

Restlessness 0 0 0 0 1 1 0 0 0 2 2


Fearfulness 0 0 0 0 0 0 0 0 0 0 0

Motor activity


Reactivity 4 4 4 4 5 5 4 4 6 6 6

Touch Response 4 4 2 0 0 0 4 0 0 0 0

Pain Response 4 4 2 2 0 0 4 0 0 0 0

Neurological

CNS excitation


Straub Tail 0 0 0 0 0 0 0 0 0 0 0

Tremors 0 0 0 0 0 0 0 0 0 2.3 2

Twitches 0 0 0 0 0 0 0 0 0 0 0

Convulsions 0 0 0 0 0 0 0 0 0 0 0


Limb Position 4 4 4 4 4 4 4 4 4 4 4


Staggering Gait 0 0 0 0 0 0 0 0 0 0 0

Abnormal Gait 0 0 0 0 0 0 0 0 0 0 0

Righting Reflex 0 0 0 0 0 0 0 0 0 0 0


Grip Strength 4 4 4 4 4 4 4 4 4 4 4

Body Sag 0 0 0 0 0 0 0 0 0 0 0

Body tone 4 4 4 4 4 4 4 4 4 4 4

Abnormal tone 4 4 4 4 4 4 4 4 4 4 4

Reflexes

Pinna 4 4 4 4 4 4 4 4 4 4 4

Corneal 4 4 4 4 4 4 4 4 4 4 4

I.F.R 4 4 4 4 4 4 4 4 4 4 4


Pupil Size 4 4 4 4 4 4 4 4 4 4 4


Exophthalmoses 0 0 0 0 0 0 0 0 0 0 0

Urination 0 0 0 0 0 0 0 0 0 0 0

Salivation 0 0 0 0 0 0 0 0 0 0 0

Piloerection 0 0 0 0 0 0 0 0 0 0 0

Hypothermia 0 0 0 0 0 0 0 0 0 0 0

Skin color 4 4 4 4 4 4 4 4 4 4 4

Heart rate 4 4 4 4 4 4 4 4 4 4 4


Lacrimation 0 0 0 0 0 0 0 0 0 0 0

Dead

Number acute 0 0 0 0 0 0 0 0 0 0 0

Number delayed 0 0 0 0 0 0 0 0 0 0 0


124

Table 5.7 Comprehensive behavioral and neurological profile of the drugs (Peak Response at highest dose)

Profile (Basal score) PG HJ CD KL IK BC

Awareness (4) 2 2 2 3 6 5

Visual Placing (4) 4 2 2 4 4 4

Stereotypy (0) 0 0 0 0 0 2

Grooming (4) 4 4 4 4 6 5

Restlessness (0) 0 0 0 0 2 2

Irritability (0) 0 0 0 0 2 2

Spontaneous Activity (4) 2 2 2 2 6 6

Reactivity (4) 2 2 2 2 6 6

Touch Response (4) 0 2 0 2 0 0

Pain Response (4) 0 2 0 2 0 0

Tremors (0) 0 0 2 0 1 2

Convulsions (0) 0 0 2 0 0 0

Righting Reflex (0) 0 0 2 0 0 0

Grip Strength (4) 4 2 2 4 4 4

Pinna Reflex (4) 0 2 2 4 4 4

Corneal Reflex (4) 1 2 2 4 4 4

Ipsilateral Flexor Reflex (4) 2 4 2 4 0 4

Death (0) 0 0 1 0 0 0

PG-Panchagavya Ghrutham, HJ-Hab-e-Jund, CD-Cynodon dactylon, KL-Kushmanda Lehyam, IK-Itrifal Kishneezi, BC-Barleria cristata.

5.4 DISCUSSION

PG, HJ and CD showed CNS depression, besides PG and CD

showed significant analgesic activity. CD showed severe tremors and

convulsions at higher doses indicating hypoglycemic activity. With these

observations further studies for antiepileptic activity were taken up.

125

KL, IK and BC showed CNS stimulation. But KL at higher doses

showed CNS depression. KL, IK and BC showed potent analgesic activity.

IK increased the volume and frequency of defecation. The drugs were

studied for their antidepressant activity.

5.5 CONCLUSION

From the results of acute toxicity studies and neuropharma-

cological profile study the doses of the drugs were decided to proceed for

further studies.

Documents

5. NEUROPHARMACOLOGICAL PROFILE (IRWIN S METHOD)shodhganga.inflibnet.ac.in/bitstream/10603/19320/13/13... · 2018. 7. 9. · Effects of the test substance were scored on a scale of