antiinflammatory lab 5

8/22/2019 Antiinflammatory Lab 5

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Anti-inflammatory

Drugs# Lab 4 #


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Inf lammat ion:

it is a biological response of vascular

tissues to harmful stimuli, such aspathogens, damaged cells, or

irritants.

Inflammation may ends with either :

Complete healing

of tissues

Permanent destruction

of tissues


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Signs of inflammation

Redness:

Due to vasodilatation by effects of

releasing of histamine, bradykininand prostaglandin.

Hotness:

Due to increased blood flow.


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Signs of inflammation

Swel l ing:

due to increased vascularpermeability by the releasedmediators and increased the exudatein the inflammed area

Pain:due to irritation of nerve ending byinflammation and the pressure of the

swelling on the nerve ending.


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Inf lammatory mediators :

histamine

5-HT (serotonin) Bradykinin

Prostaglandins (eg PGE2 )

Interleukines

Substance P

Nitrous oxide

Main

inflammatory

mediator


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Phases of

Inflammation

Flu id phase (vascu lar phase):

Increased vasodilatation leads to

increased permeability of the vascular

bed to plasma protein.

- Increasing fluid will help in:1- dilution of the irritant

2- increase conc. of antibodies from blood

to inflamed area

3- supply nutrients to the immune cells.


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Cellular phase (exudative phase):

Involves migration of tissue

macrophages and polymorphonuclearleukocytes (PMNL) to the inflamed area.

Fibrous phase (p ro l i ferat ive phase):

A new connective tissue (fibrous)containing fibroblast and capillaries is

formed.

Phases of

Inflammation


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Classification of the

InflammationNonimmunological :

Induced by chemical irritants such as formalin

Immunological :Induced by infections such as bacterial infection


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Anti-inflammatoryDrugs

Steroidal Non-steroidal

- Cortisone

- Hydrocortisone- Acetaminophen

- Aspirin


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Steroids (SAIDs)

- Containing steroid moiety in their sturcure

Glucocorticoids (GC)

Cortisone


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Glucocorticoids (GC)

Natural Synthetic

- Cortisone

- Hydrocortisone

- Betamethasone

- Dexamethasone

- Predinsone

Fluorinated

Glucocorticoids

Prednisolone

Liver enzymes


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Glucocorticoids (GC)Mechanism of Action :

They act by indirect inhibition of the enzyme phospholipase A2

which activate synthesis ofarachidonic acid with subsequentformation of prostaglandins.

They induce synthesis of a protein lipocortin-1 which has

the inhibitory effect on phospholipase A2.

-

-


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phospholipase A2

GC inhibition


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Side Effects : Immunosuppression

Hyperglycemia due to increased gluconeogensis, insulin resistance, and

impaired glucose tolerance ("steroid diabetes");

Steroid-induced osteoporosis: reduced bone density (osteoporosis,

Osteoporosis , higher fracture risk, slower fracture repair)

Redistribution of body fat: moon face, buffalo hump and truncal obesity.

Adrenal insufficiency

Muscle breakdown (proteolysis), weakness; reduced muscle mass and repair

Anovulation, irregularity of menstrual periods

Growth failure, pubertal delay Increased plasma amino acids, increased urea formation;

Glaucoma due to increased cranial pressure


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Side Effects :

Moon face buffalo hump


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Non-Steroidal Anti-inflammatory Drugs

(NASID)

They dont contain steroid moiety

They also have analgesic and antipyretic activity


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Mechanism of Action :

NSAIDs inhibit synthesis of PGs which are the main factors

Playing a role in the inflammaltion.

Inhibit synthesis of PGs through inhibition of cyclooxygenase

Enzymes which are responsible for production of PGs


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GC inhibition

phospholipase A2


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Cyclooxygenese ( COX ) Isoforms :

COX 2COX 1

- Constitutive

- Many tissues ( blood vessels

stomach and kidney )

- inducible

-By inflammatory processes

and mediators

COX 3 has recently been described


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Non-Steroidal Anti-inflammatory Drugs

(NASID)

Non-selective

COX inhibitors

selective COX2

inhibitors- Aspirin

- Ibuprofen

- Diclofenac

- Meloxicam

- Celecoxib

- Rofecoxib


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Side Effects :Unwanted effects, owing largely to inhibtion of COX1

Particularly in the elderly and include :

- Despepsia, nausea and vomiting , ulceration and gastricdamage in chronic users, with risk of hemorrhage

- Reversible renal insufficiency

- Analgesic-associated nephropathy ( irreversible )

- Liver disorders, in high doses, e.g. acetaminophen


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Measurement the activity

ofanti-inflammatory drugs

- Method : Paw Oedema Method

-Principle : induction a chemical inflammationby injecting an irritant ( formalin ) into

rats paw

-Objective : measure the anti-inflammatory activity ofdiclofenac and hydrocortisone with

different doses )


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-Procedure :

1- select 5 rats2- inject each rat 1 ml urethane for anesthesia.

3- select one as control and inject the rest of them intraperitoneal

rat 1 >>> control

rat 2 >>> 40 mg/kg diclofenacrat 3 >>> 80 mg/kg diclofenac

rat 4 >>> 20 mg/kg hydrocortisone

rat 5 >>> 40 mg/kg hydrocortisone

4- after 1 hr , inject 0.1 ml formalin in each rat ( 2 to 5 ) into

their paws >>> to induce inflammation.

5- after 1 hr , take the reading using the plythysmometer of each

rat paw ( right and left ).

6- calculate the inflammation and response % for each drug.


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Response

%

inflammat

ion

LPRPDose

___Ccontrol

T140 mg/kg vol.

T280 mg/kg vol.

T320 mg/kghydro.

T440 mg/kg

hydro.Inflammation = RP - LP

Response % = X 100C - T

C

Response % >>>> Anti-inflammatory activity

antiinflammatory lab 5

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