Anti-Inflammatory Activity Of Canthium Angustifolium Roxb Leaves By Carrageenan Induced Paw Oedema And Cotton Pellet Granuloma Method

Inflammation is a protective, response involving host cells, blood vessels, and proteins and other mediators that is intended to eliminate the initial cause of cell injury, as well as the necrotic cells and tissues resulting from the original insult, and to initiate the process of repair. Inflammation can be acute or chronic. Acute inflammation is rapid in onset and of short duration, lasting from a few minutes to as long as a few days, and is characterized by fluid and plasma protein exudation and a predominantly neutrophilic leukocyte accumulation. Chronic inflammation may be more insidious, is of longer duration (days to years), and is typified by influx of lymphocytes and macrophages with associated vascular proliferation and fibrosis (scarring)1. Inflammation results in the liberation of endogenous mediators like histamine, serotonin, bradykinin, prostaglandins etc. These mediators even in small quantities can elicit pain response.  The greatest disadvantage in presently available potent synthetic drugs for the treatment of inflammation lies in their toxicity and reappearance of symptoms after discontinuation. Therefore the screening and development of drugs for their anti-inflammatory activity is still in progress and there is much hope for finding anti-inflammatory drugs from indigenous medicinal plants. There is a growing interest in the pharmacological evaluation of various plants used in Indian traditional systems of medicine.2

Canthium angustifolium Roxb is extensively used in Indian traditional medicine for several diseases including inflammatory disorders but there is no scientific evidence available for such activity. Based on the traditional uses the aim of present study was to investigate the anti-inflammatory activity of leaf extract of Canthium angustifolium Roxb on wistar rats.

MATERIALS AND METHODS

Plant material:

The plant material was collected from Cheruvandoor, Kerala and taxonomic identification of sample was confirmed by C.M.S College, Kottayam (Voucher specimen no:274).

       
           
       
    Fig 1: Canthium angustifolium

Preparation of Plant extract:

Fresh leaves were collected and dried at room temperature to remove moisture, and size reduced. Extraction of the dried leaves of Canthium angustifolium Roxb was carried out by soxhalation by ethanol (TEE).

Animals used:

Female albino Wistar rats (220-250)g and Male Wistar albino rats (150-200)g were housed in standard environmental conditions. Food and water were available ad libitum. Female albino Wistar rats were used for the acute toxicity study in accordance with OECD guidelines 423. Male Wistar albino rats were used for screening anti-inflammatory activity of Canthium angustifolium Roxb. All the experiments were done after approval by the IAEC (Institutional animal ethical committee) of College of Pharmaceutical Sciences, Cheruvandoor. (IAEC No: 020/MPH/UCP/CVR/14).

Carrageenan induced paw oedema:

Male Albino wistar rats with a body weight between 150-200g were used. The animals were starved overnight, water given ad libitum. Group I receives vehicle (CMC), Group II receives Diclofenac Na 10mg/Kg and group III and group IV receives 200mg/Kg and 400mg/kg of total  ethanolic extract of Canthium angustifolium Roxb respectively. Thirty minutes later, the rats were challenged by subcutaneous injection of 0.1ml of 1% solution of carrageenan in saline into the plantar region of the left hind paw. The paw was marked with ink at the level of lateral malleolus. The readings were measured by immersing the paw in mercury upto the mark. The paw volume were measured plethismographically immediately after the injection and again 1, 2 and 3hour after challenge.

Percentage inhibition of paw edema =  (Vt-V0)control - (Vt-V0)test  ? 100(Vt-V0)control

Vt is the rat paw volume at time t, V0 is the initial rat paw volume (before carageenan injection), (Vt-Vo) is edema produced in control group and (Vt-V0) treated is edema produced in treatment groups. The difference in the average values between treated animals and control groups were calculated for each time interval and statistically evaluated3.

Cotton pellet granuloma:

The rats were anaesthetized with diethyl ether and a sterilized cotton pellet weighing approximately 10±1mg was inserted in the subcutaneous layer of groin. The incised skin was properly sutured. The control group receives vehicle (CMC); Standard group receives dexamethasone (0.5 mg/Kg) p.o; Test group animals were treated with the total ethanolic extract of Canthium angustifolium Roxb (200mg/Kg & 400mg/kg p.o) once a day for 7 continuous days. On 8th day animals were sacrificed by cervical dislocation and the pellets along with granuloma mass were removed, washed and taken the wet weight. Then the granuloma mass were dried at 60°C for 24 h in an oven and taken the dry weight. On 0th day and 8th day (before sacrificing) blood was withdrawn from each animal by retro orbital puncture for evaluating the leucocyte count4. Weight of the granuloma mass was calculated as percentage and compared the weight of treated group with control group.

% inhibition = Wc-WdWc ? 100

Wc – Difference in pellet weight of control

Wd- Difference in pellet weight of treated group

Statistical analysis

 All data were expressed as mean ± SEM and analyzed by One-way analysis of variance (ANOVA) followed by Dunnet’s test. P< 0>

       
           
       
    Table 1:  Anti-inflammatory activity study using carrageenan induced paw oedema model

test. *p< 0>

       
           
       
    Graph2 : Effect OF TEE on wet granuloma weight:

test. *p< 0>

       
           
       
    Graph 4 : Effect on leucocyte migration

RESULTS  AND DISCUSSIONS