Lakhasly

ABSTRACT This work aims at evaluating the anti-inflammatory activity of an endemic species of the central sahara: Myrtus nivellei Batt & Trab.In sight of this, our attention has been targeted to Myrtus nivellei Batt & Trab, an endemic Saharan species, belonging to the family of Myrtaceae, which grows in scattered populations, in rocky and sandy wades where subterranean water points exist, and generally at an altitude above 1800m [3].It is a shrub up to 2 meters hight, with rough bark, leaves lanceolate, thick and linear (4 to 5 cm), five white petals, indeterminate stamens and the fruits are black berries [4,5].In the carrageenan-induced paw edema test, five different groups of mice were established and the extract was administered orally in three different doses.The dose of 400 mg/kg was able to reduce significantly the paw edema with a comparable effect to that observed with Diclofenac (positive control).Keywords: Anti-inflammatoy activity; Myrtus nivellei Batt & Trab; methanolic extract; paw edema.The methanolic extract of this plant was extracted by Soxhlet apparatus and concentrated under reduced pressure using a rotary evaporator.INTRODUCTION In recent years, there has been a considerable interest on the secondary metabolites obtained from many plants which gained a big scientific interest.This is the first report to demonstrate a significant anti-inflammatory activity of the methanolic extract prepared from Myrtus nivellei.Consequently, the search for new anti-inflammatory compounds has been a priority for the pharmaceutical industry.Many natural products have been tested in various animal models for the development of new anti-inflammatory agents [2].Medicinal plants continue to be an important source of new chemical substances with potential therapeutic effects [1].Commercially available anti-inflammatory drugs exert a wide range of side effects and are either too potent or too weak.1.

ABSTRACT
This work aims at evaluating the anti-inflammatory activity of an endemic species of the central
sahara: Myrtus nivellei Batt & Trab. The methanolic extract of this plant was extracted by
Soxhlet apparatus and concentrated under reduced pressure using a rotary evaporator. In the carrageenan-induced paw edema test, five different groups of mice were established and
the extract was administered orally in three different doses. The dose of 400 mg/kg was able to
reduce significantly the paw edema with a comparable effect to that observed with Diclofenac
(positive control). This is the first report to demonstrate a significant anti-inflammatory activity
of the methanolic extract prepared from Myrtus nivellei. Keywords: Anti-inflammatoy activity; Myrtus nivellei Batt & Trab; methanolic extract; paw
edema.

1. INTRODUCTION
   In recent years, there has been a considerable interest on the secondary metabolites obtained
   from many plants which gained a big scientific interest. Medicinal plants continue to be an
   important source of new chemical substances with potential therapeutic effects [1].
   Commercially available anti-inflammatory drugs exert a wide range of side effects and are
   either too potent or too weak. Consequently, the search for new anti-inflammatory compounds
   has been a priority for the pharmaceutical industry.Many natural products have been tested in various animal models for the development of new
   anti-inflammatory agents [2].
   In sight of this, our attention has been targeted to Myrtus nivellei Batt & Trab, an endemic
   Saharan species, belonging to the family of Myrtaceae, which grows in scattered populations,
   in rocky and sandy wades where subterranean water points exist, and generally at an altitude
   above 1800m [3]. It is a shrub up to 2 meters hight, with rough bark, leaves lanceolate, thick
   and linear (4 to 5 cm), five white petals, indeterminate stamens and the fruits are black berries
   [4,5]. It’s known under the names of “Tafeltest” in Tamahaq and “Raihane Essahara El
   Wousta” in Arabic. Very little is known about the phytochemistry and biological activities of
   Myrtus nivellei. To our knowledge there is only few papers reporting the phenolic compounds
   and its antioxidant activity [6,7] and there is none about the potential activities of Myrtus
   nivellei extracts.
   The purpose of the present study was to evaluate the anti-inflammatory activity of the methaolic
   extract using the carrageenan-induced paw edema test, for enduring safety and exploring the
   beneficial role of this plant to treat inflammatory diseases.


2. MATERIAL AND METHODS
   2.1. PLANT MATERIAL
   Aerial parts of Myrtus nivellei were collected in July 2013 during the flowering stage. near to
   Tamanrasset city (Hoggar massif, altitude: 1900 m, latitude: 22°38, longitude: 5°37). The plant
   was identified as Myrtus nivellei at the department of Botany INA-El Harrach (Algeria). The
   fresh leaves were washed thoroughly to remove dirt and dried under shade for a week. They
   were grounded into fine particles and stored until use.
   2.2. EXTRACTION AND SAMPLE PREPARATION
   Ground samples of Myrtus nivellei were weighted and extracted with methanol using a Soxhlet
   apparatus for 6 hours [8]. Then, the extract was concentrated to dryness under reduced pressure
   using a rotary evaporator to obtain a methanolic dry extract.
   The dry extract was dissolved in a vehicle solution (1% Tween 80 in distillated water), and
   sterilized by filtration through sterile syringe filter with 0.2µm pore. Finally the filtered extract
   was stored in glass bottles at +4°C until use.2.3. EXPERIMENTAL ANIMALS
   Carrageenan-induced paw edema was carried out on male Swiss mice (25−30 g). Male animals
   were purchased from the laboratory of toxicology of Antibiotical Company (Medea, Algeria)
   and were housed in groups of six per standard cage, on a 12 h light/dark cycle with free access
   to food and water. They were acclimatized to laboratory conditions for at least 1 week before
   testing. The food was withdrawn on the day before the experiment, but free access to water was
   allowed. A minimum of six animals was used in each group.
   2.4. DETERMINATION OF MEDIAN LETHAL DOSE (LD50)
   LD50 of the extract was estimated in mice by using the method of Hilan et al [9]. In a preliminary
   test, animals in groups of three received 10, 100, or 1,000 mg/kg of dry methanolic extract
   suspended in the vehicle (1% v/v Tween 80) and sterilized by filtration through sterile syringe
   filter with 0.2µm pore. Animals were observed for 24 h for signs of toxicity and number of
   deaths. The LD50 was calculated as the geometric mean of the dose that resulted in 100%
   mortality and that which caused no deaths.
   2.5. IN VIVO ANTI-INFLAMMATORY ASSAY: CARRAGEENAN-INDUCED PAW
   EDEMA IN MICE
   The anti-inflammatory activity was evaluated by the carrageenan-induced paw edema test [10].
   Paw edema was induced by injecting 0.1 ml of the carrageenan 1% suspension in isotonic saline
   (w/v) into the sub-plantar region of the left hind paw of the mouse.
   The doses of 100, 200, or 400 mg/kg and vehicle (0.2% Tween 80 in 0.9% NaCl) were
   administrated orally (per os) 30 min before injection of the edematogenic agent to different
   groups of mice for each treatment (n=6 per group). Diclofenac sodium dissolved in 0.9% NaCl
   (50 mg/kg, oral) was used as a reference drug. Paw thickness was measured before the
   application of the inflammatory substance and every 30 min for 4 h after induction of
   inflammation. The difference in footpad thickness was measured by a gauge calliper (Facom,
   Paris, France).
   Mean values of treated groups were compared with those of control group (vehicle) and
   analyzed statistically. The data obtained for the various groups are reported as means±standard
   deviation (SD) and expressed in mm.
   The percentage inhibition of the inflammatory reaction was determined for each animal by
   comparison to the controls and calculated by the formula:I(%) = 1 −
   ∆(PV)t
   ∆(PV)c
   x100
   Where I (%)=percentage inhibition of edema, ( PV)t=the change in paw volume in the
   treated mice, and ( PV)c=the change in paw volume in the control mice.
   2.6. STATISTICAL ANALYSIS
   Results of the paw edema of the mice are reported as mean±SD. Comparison between groups
   was made by one-way analysis of variance (ANOVA). Differences with P