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After treatment with adrenocortical hormones, rats are put in metabolic cages to collect urine through a funnel.
Antiinflammatory effects
Granuloma test in rats Eosinopenia test in mice Rat-paw edema
Antiinflammatory effects
cotton pellets are implanted in the axillae to produce granuloma. The pellets will be considered as foreign bodies leading to an inflammatory reaction. After 1 week, the cotton pellets become surrounded by fibrous tissues (granuloma), they are isolated from the animal and weighed (pellet + granuloma). The animal is given the glucocorticoid preparation and the procedure is repeated then the granuloma weight is decreased
Antiinflammatory effects
Eosinopenia test in mice:
glucocorticoids reduce the number of eosinophils in a proportional way to the dose
Antiinflammatory effects
Rat-paw edema
(carrageenan-induced edema in the rat hind paw) Carrageenan is inflammatory materials used to induce experimental inflammation due to the release of inflammatory mediators (histamine, kinins and prostaglandins).
Antiinflammatory effects
Rat-paw edema
Carrageenan sodium gel (5 micro ml of 1% solution) is injected S.C. in the rat hind paw causing inflammation and edema after 1 hour. The volume of edema in the hind paw is measured by volume displacement using mercury plethysmography. The hind paw is placed in mercury, causing mercury displacement. The displaced volume is measured by mercury manometer.
Antiinflammatory effects
Rat-paw edema
Another group of rats is injected I.P. by the anti-inflammatory
drug then, after 1 hour, carrageenan are injected into the hind paw. Anti-inflammatory drugs reduce the volume of edema and displaced mercury.
Test drug I.P Carrageenan S.C
Antiinflammatory effects
Rat-paw edema
S1
S2
T1
T2
Procedure
12 to 20 healthy rabbits (1.5 to 2 kg) are used. A preliminary test is carried out to exclude any rabbit liable to convulsion, where rabbits are fasted for 18 hours then each rabbit is injected s.c. with one unit of insulin and observed for 5 hours; any rabbit showing convulsions is excluded.
are randomly distributed into 4 groups and deprived of food for not less than 18 hours before carrying out the assay.
Blood samples are withdrawn from the marginal ear vein and the average concentration of blood glucose in mg/dl is determined in each group (initial blood sugar). A 4-point assay is performed using the 4 rabbit groups, where 2 doses of the test and 2 doses of standard insulin preparation are injected so that the dose of insulin does not exceed 0.5 unit/kg body weight.
S1
S2
T1
T2
Following insulin injection, blood samples are withdrawn from the marginal ear vein every hour for 5 consecutive hours and the average concentration of blood glucose in mg/dl is determined in each group (final blood sugar). For each dose of the test and standard insulin, the reduction in blood sugar level is calculated and the potency of the test insulin is determined.
Bioassay of Analgesics
Mechanical methods
Thermal methods
Principle: The mouse tail (as well as the ear) is not covered by hair so it is very sensitive to pain due to the presence of sensory nerves in the skin.
Control
An artery clip (clamp) is applied to the base of the mouse tail for 30 seconds. The animal will feel pain and will try to get rid of the clip A group of mice is treated with the test drug, and the clip is applied to the tail after 30, 60,120 and 180 minutes If the animals response to the clip is abolished or delayed,then the drug may have analgesic properties. The ED50 is calculated
Test
*Principle:
Electric shock
*The rats tail is used, and an electric current is allowed to pass through a coil until it becomes bright red. *The reaction time, which is the time elapsed between closing of electric circuit and withdrawal of tail, is recorded.
irritation stretching & writhing (twist due to pain) count no. of writhings -Give Aspirin then glacial acetic acid count no. of writhings
Writhing method
3.) Procedure: 1. Weigh mouse 2. Inject mouse IP with 1ml/100 g of 10% glacial acetic acid 3. Record the onset and the count of writhings 4. Give Aspirin orally to another mouse in a dose of 50 mg/kg 5. After 30 min inject glacial acetic acid IP 6. Record the onset and count of writhings and compare