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INSTITUTE OF PHARMACOLOGY
Experiment 1 The basic knowledge and technique of pharmacological
experiment
2. A drug passes through membranes more readily if it uncharged. AABB increase absorption;
ABBA accelerate excretion.
pKa-pH=lg(BH+ / B)
3. Strychnine is a weak basic CNS stimulant. It can excite spine selectively and cause convulsion
(legs or limbs are rigid), even die.
Materials .
3. animals: 24 mice
4. drugs: strychnine solution (pH =8.0 or 1.0); picric acid solution.
Methods
5. weigh the animals and stain them
6. animal grouping randomly: 12 mice per group
7. drug administration: ig 0.3ml/10g, record the time after administration.
8. observe the effects of every mice and the time each mice appearing convulsion within 15min
after drug administration
Results:
group n number of convulsive mouse The time each mouse appearing
convulsion
AIM:
1. to observe the effect of acetylcholine, atropine and BaCl2 on smooth muscle of ileum and
analyze the mechanism of action of atropine.
2. to grasp how to do experiment in vitro
Principles
1. M-R: distribution and effect when M-R is activated
To the ileum, the effect when M-R was stimualted is contraction of smooth muscle and increase
of tension of ileum.
Ach is an agonist of M-R, while atropine is an antagonist of M-R, so it can antagonize the
effect of Ach.
MATERIALS
1. drugs: Ach, atropine, BaCl2
2. animal:guinea pig
3. instruments: biolap98, tonotransducer,a series of eqiupments for experiment in vitro
Methods
1. set the temperature at 32 degree
2. setup the software named“biolap98”
enter “biolap98”---- choose number 1 channel----press “input signal”- “channel 1” --
“tension”----press “start”.
Press a button to let the curve lower 5 divisions from the middle line
set two parameters:(1)amplification mutiple: 100 (2)scan speed: 25s/div
3. measure 20ml of tyrode’s solution and pour it into the bathing tube
4. prepare the samples: 2cm ileum
get a guinea pig, and beat its head, when it dies, open its abodemen and get its ileum out, wash it
with tyrode’s solution, and cut it into several part, the lengh of each section is about 2cm.
Be careful when prepare the samples. Do not hurt the ileum with too much force .
5. hang the sample diagonally . suspend the sample quickly and incubated it with tyrode’s solution
and air. The air bubble 2-4 every second
Sample: one end on L type hook, the other end on thread hook. Fix the L type hook on iron stand,
fix the thread on tonotransducer. Let the thread not too loose, not too tense.
6. adjust the load: adjust the height of L type hook and let the curve ascending two divisions and fix
the L type hook.
7. add the drugs to the bathing tube : 0.2ml each time.
The sequence is : Ach---- Atropine-----Ach------ BaCl2-----Atropine
First add Ach, when the effects of Ach reach its peak, add Atropine, when the curve returned to
stable, add Ach for another time. Add BaCl2 , when its action reach peak, add Atropine .
8. stop the experiment and save the results.
RESULTS
Disscusion: analyze the results.
.
Experiment 6 Anticoagulant Effects of Drugs in Vitro
AIM:
To compare the characteristic of different anticoagulant drugs and analysize their mechanism.
Principles:
Heparin enhance the effect of antithrombin III to inactivate the coagulant factor 2,
9,10,11,12,so it can cause anticoagulant effect in vivo and in vitro.
Dicoumarin inhibit the modification or synthesis of the coagulant factor 2, 7, 9,10 in liver.
Sodium citrate solution always is used in storing blood, which produces anticoagulant effect by
combining calcium.
MATERIALS
drugs: Heparin,Dicoumarin,Sodium citrate solution,normal saline, potassium oxalate solution,
calcium chloride solution
animal: rabbit
Methods
3. Mark the tubes 0, 1, 2, 3.
Add drugs: add 0.25ml drugs into every tube respectively.
0----NS; 1---- Dicoumarin; 2----Heparin; 3----Sodium citrate solution
2. Anesthetize rabbit.
3. Operation: artery catheterization.
4. Get 5ml blood by injector washed by potassium oxalate solution and put 0.9ml blood into every
tube, then add 3mg/ml calcium chloride solution 0.1 ml at once, and after mixing gently, put the
tubes into 37centigrade water bath, record the time.
5. Observe: slope the tubes every 30 second gently. If the blood stops flowing, it is coagulant.
Please record the time.
6. If the blood in the tubes will never appear coagulant, add another high concentration 10mg/ml
3mg/ml calcium chloride solution 0.1 ml, go on observing just like before.
RESULTS
Disscusion: analysize the results.
Experiment 7 Effects of drugs on arterial blood pressure of anesthetic rabbit
AIM: to observe the effects of autonomic drugs on rabbit arterious blood pressure
Principles
1. heart: M-R and β1-R
vascular smooth muscle: α-R andM-R, β2-R
2. autonomic drugs include drugs acting on cholinoceptor and drugs acting on adrenoceptor. These
drugs exhibit their effcts through binding to receptors.
Materials:
1. animal: rabbit
2. drugs : Ach, neostigmine, atropine, NA, AD, Isop, phentolamine, propranolol
3. eqipmemts: BL-410, operating instruments, artery catheter, trachea catheter, pressure transducer
Methods
1. setup the software named“BL-410”
enter “BL-410”---- choose number 1 channel----press “input signal”- “channel 1” -- “tension”----
press “start”.
2. anesthetize the rabbit: 20% urathene 5~6ml/kg
3. operation:
(1) trachea catheterization
(2) CCA catheterization
(3) Fix the scalp needle ( for drug administration )
4. Drug administration: iv through the scalp needle
Group A
(1) Ach 0.1ml/kg
(2) Neostigmine 0.1ml/kg
(3) Ach 0.1ml/kg
(4) Atropine 0.4ml/kg
(5) Ach 0.1ml/kg
Group B
(1) AD 0.1ml/kg
(2) NA 0.1ml/kg
(3) Isop 0.1ml/kg
(4) Phetolamine (regitine) 0.2ml/kg slowly
(5) 5min later, repeat (1)(2)(3)
Group C
(1)AD 0.1ml/kg
(2)NA 0.1ml/kg
(3)Isop 0.1ml/kg
(4)propranolol 0.1ml/kg slowly
(5)5min later, repeat (1)(2)(3)
Results:
Print the figure and analyze the effects of the drugs
Experiment 8 Effects of Drugs on the isolated Guinea-pig Tracheal Ring
AIM:
To observe the effects of drugs on tracheal smooth muscle and select the asthmatic drugs.
Principles:
3. There are M-R, H1-R, β2-R in tracheal smooth muscle.
4. When M-R and H1-R are stimulated, the tracheal smooth muscle will be contraction and the muscular tension
will be increased.
When β2-R was stimulated, the tracheal smooth muscle will be relaxation and the muscular tension will be
decreased.
5. The conditions for an isolated tracheal ring to be alive in vitro:
(1) temperature: 37 ℃
(2) gas: oxygen
(3) nutrient solution: Krebs-Henseleit’s solution
MATERIALS
4. drugs: histamine, isoprenaline, Ach, aminophyline, propranolol
5. animal: guinea pig
6. instruments: bl-410E software, tonotransducer,a series of eqiupments for experiment in vitro
Methods
4. Set the temperature at 37 centigrade.
5. Start the software names“BL-410E”
enter “BL-410E”---- choose number 1 channel----press “input signal”-“channel 1” -- “tension”----press
“start”.
set parameters: amplification mutiple: 20
3. Measure 15 ml of Krebs-Henseleit’s solution and pour it into the bathing tube
4. Prepare the samples: 0.5cm tracheal, get a guinea pig, and beat its head, when it dies, get the tracheal out, and
cut it into several part, the lengh of each section is about 0.5 cm. Be gentle when prepare the samples. Do not
hurt the ileum with too much power.
5. Hang the sample parallel. Hang the sample quickly and incubated it with K-H’s solution and oxygen, Make the
air bubble continuously (2-3 every second).
Sample: one end on L type hook, the other end on thread hook. Fix the L type hook on iron stand, fix the thread
on tonotransducer. Let the thread not too loose, not too tense.
6. Adjust the load: adjust the height of L type hook and let the curve ascending 2g and fix the L type hook.
7. Add the drugs to the bathing tube: 0.15 ml each time. Firstly, stabilize 15 min
The sequence is:
Ach (quiescently scan) ---- reach its peak (need about 5min-10min, then scan 1cm distance.)----Isop (like
before) clean 3 times, stabilize 10 min
HA-----Amino
Prop------ Isop-----Amino
8. Stop the experiment and save the results.
RESULTS
Disscusion: analyze the results.