Screening of Analgesic and Anti-inflammatory Activity of Hydroalcohol Extract of Sida (Indian) Species Root

VH Bhaskar1, B Sangameswaran1, N Balakrishnan2*, AB Panda2, Navin R Raj2 and A Sathish3

1 Faculty of Pharmacy, Vinyaka Mission's University, Salem-8, Tamilnadu.

2Technocrats Institute of Technology – Pharmacy, Bhopal-462021 (MP).

3 A. R. College of Pharmacy, Anand, Guajarat.

* Corresponding Author E-mail: pharmacobala@gmail.com

ABSTRACT

The Sida acuta, Sida cordifolia, Sida rhombifolia and Sida spinosa (Malvaceae) and are considered to be one of the most valuable drugs in Ayurvedic medicine. All the parts of these plants was highly useful and among them root seems to be much useful. Traditionally Sida species used as externally healing of wounds, skin ulcers, inflammation and internally used in treatment of fever, liver disorders, analgesic. The Sida species distributed throughout the tropical and sub-tropical regions of the world and all over India. The anti-inflammatory activity of Sida species root extract were evaluated by carrageenan induced rat edema model and analgesic activity was determined by acetic acid induced writhing method in mice. In the entire models, group I served as control, group II, III, IV and V received extract at the dose level of 200 mg / kg body weight respectively and group VI served as standard drug. The results revealed that hydroalcohol extract of Sida cordifolia roots possess potent anti-inflammatory and analgesic activity among the four Sida species.

KEY WORDS Sida acuta, Sida cordifolia, Sida rhombifolia, Sida spinosa.

INTRODUCTION:

In Indian system of medicine, a large number of drugs of either herbal or mineral origin have been advocated for various types of diseases and other different unwanted conditions in humans. Ayurveda is one of the traditional systems of medicine practiced in India and Sri Lanka can be traced back to 6000 BC. Ayurvedic medicines was largely based upon herbal and herb mineral and have specific diagnostic and therapeutic principle1. The medicinal values of Sida species have been known from ancient times and are currently used to treat various ailments. The plant Sida acuta, Sida cordifolia, Sida rhombifolia and Sida spinosa (Malvaceae) are used treatment for externally healing of wounds2, skin ulcers, inflammation and internally used in treatment of fever, liver disorders, analgesic3.

Plant Description

Sida acuta is a low shrub to 1 m tall or rarely 2 m with slender tough stems. Leaves lanceolate or linear- lanceolate, acute, obtuse or rounded at base, 1.5-6 cm long, 5-20 mm wide, sharply serrate, stellate-pubescent beneath, petioles 3-5 mm long, blade 3-nereved from base, pinnately branched above the base, stipules linear-acute, about 1 cm long. Flowers 1 or 2 in axils, pedicels to about 1 cm long, corolla about 6-14 mm broad and yellow in colour. Fruit 4-5 mm broad, 6-8-carpellate and seeds dark reddish-brown, 1.5 mm long, ripe carpels with 2 beaks, grooved on the back.

Sida cordifolia is an erect subshrubs 0.5-1.5m tall, whitish to yellowish stellate velutinous or stellate tomentose. Leaf blades ovate to oblong or sometimes orbicular, 2-10 cm long, margins serrate, base usually shallowly cordate, petioles usually 1/3 or more the length of blades, stipules filiform and 3-7 mm long. Flowers usually in axillary clusters, often forming dense panicles or corymbs of racemes by reduction of leaves, calyx 5-7 mm long, accrescent and clasping in fruit, petals cream to pale yellow or orange yellow, 7-9 mm long, staminal column included. Schizocarp 6-7 mm in diameter, mericarps 8-12, brownish, the body 2.8-3.5 mm long, with 2 retrorsely simple pubescent apical awns 2-4 mm long, dorsal and lateral surfaces rugose-reticulate. Seeds are 1.8-2.3 mm long, glabrous or with a few hairs along the raphe.

Sida rhombifolia is small, tough-stemmed shrub to 1m tall, stellate-pubescent, stipules linear, 6-9 mm long. Leaves ovate or rhombic-ovate, acute, serrate near the apex, glabrous above, pubescent beneath, 1.5-5 cm long, 7-22 mm wide, on petioles about 2-6 mm long (rarely longer). Flowers 1 or 2 in an axils, pedicels 1-4 cm long, calyx-lobes deltoid apiculate, corolla yellow, about 6-10 mm wide, rotate, schizocarp nearly 1 cm broad, ripe carpels deltoid, apiculate, 8 or 10 or rarely to 12, stellate- pubescent dorsally, 2-lobed, about 3-4.5 mm long. Seeds are black in colour.

Sida spinosa is an erect annual or perennial herb 0.5- 0.8 m tall, copiously but minutely stellate puberulent, eventually glabrate. Leaf blades linear to narrowly oblong or ovate, 1-5 cm long, margins serrate to crenate, base obtuse to truncate, petioles usually or less the length of blades, stipules filiform, 2-5 mm long, each subtended by a short tubercle. Flowers solitary or in clusters, often somewhat corymbose toward the ends of the branches, pedicels slender, 0.2-2.5 cm long, calyx strongly 10-ribbed, 4-7 mm long, petals pale yellow to yellowish orange, 5-6- (-7) mm long, staminal column included. Mericarps 5, 3-4 mm long, grading apically into 2 antrorsely pubescent awns 0.5- 1.5 mm long, lower dorsal and lateral walls strongly reticulate, apical surfaces smooth, puberulent. Seeds are 1.5 mm long, glabrous or with a few hairs around the hilum4. The present study was undertaken to screen analgesic and anti-inflammatory activity of hydroalcohol extract of Sida species roots.

MATERIALS AND METHODS:

Plant Material

The roots of Sida acuta, Sida cordifolia, Sida rhombifolia and Sida spinosa was collected from various localities of Salem, Erode and Dharmapuri districts of Tamilnadu, India in October 2002. The plant specimen was satisfactorily identified and confirmed with help of herbarium at Botanical Survey of India, southern Circle, Coimbatore, Tamilnadu, India. The voucher specimen has been retained in our laboratory for further reference.

Preparation of Extract

All the four roots were shade dried, powdered and passed through 40 meshes and stored in closed vessel in dark place at room temperature for further use. The dried powdered material 500 g macerated for seven days with 50% alcohol with frequent shaking separately at room temperature by using maceration bottle .On the seventh day it was filtered and the solvent was completely removed under reduced pressure and a semisolid mass (SAHE-12 %, SCHE-12.6 %, SRHE-11.8 % & SSHE-12.2 % ) was obtained. The hydroalcohol extract of Sida species roots was suspended in distilled water using 0.5% carboxymethl cellulose and used for the present study.

Animals

Studies were carried out using Wister albino rats of either sex weighing 180 – 200 g and Wister albino mice of either sex weighing 18 -25 g . The animals were grouped in polyacrylic cages (38 cm x 23 cm x 10 cm) with not more than six animals per cages and maintained under standard laboratory conditions (temperature 25 ± 2 ° C) with dark and light circle (14 / 10 h). They were allowed free access to standard dry pellet diet (Hindustan Lever Ltd, India) and water ad libitum .The rats were acclimatized to laboratory condition for 10 days before commencement of experiment. All procedures described were reviewed and approved by the university animal ethical committee.

Acute Toxicity studies

Toxicity studies of hydroalchol extract were carried out in oral doses of 100 to 2000 mg / kg - body weight using albino mice. After test extracts administration, animals were observed 72 h period. The number of deaths was expressed as a percentile and the LD50 was determined by probit a test using the death percentage versus the log dose 5.

Phytochemical Analysis

The extract was screened for the presence of various constituents employing standard screening test6. Conventional protocol for detecting the presence of steroids, alkaloids, tannins, flavonoids, glycosides, etc. was used.

Evaluation of Analgesics activity

Analgesic activity was determined by acetic acid-induced writhing method 7 in mice by treating with test extracts orally at a dose of 200 mg / kg body weight. Control groups were given only carboxymethylcelllulose while other group was given with reference standard diclofenac sodium 50 mg / kg body weight under similar conditions. After 30min. of administration of extracts and standard drug, all groups of mice were given the writhing agent (aqueous acetic acid 3 %) in a dose of 2 ml / kg body weight intraperitoneally. The writhing produced was counted for 10 min. and percentage protection was calculated.

Evaluation of Anti-inflammatory activity

Anti-inflammatory activity of hydroalcohol extract of Sida species was studied by carrageenan induced rat paw edema method 8. Four group’s rats were pretreated with test extracts in dose of 200 mg / kg body weight orally. One group received only carboxymethylcellulose, which served as control, while the other group received paracetamol as standard drug for comparison in the dose of 100 mg / kg body weight under similar conditions. After 30min., 0.1 ml of carrageenan (1 %) was injected into planter region of hind paw of rat. Measurements of paw volume (ml) were made by mercury displacement technique using plethysmometer. Percentage inhibition of inflammation was calculated after 4th h.

Statistical Analysis

All pharmacological evaluation was expressed as Mean ± Standard error. Test significance was statistically evaluated by Student’s t-test 9.

Table-1 : Analgesic and Anti-inflammatory activities of Sida species

S. No.	Extract / Standard	Analgesic activity		Anti-inflammatory activity
S. No.	Extract / Standard	Writhingin10min Mean ±SEM	Percentage protection	Paw volume (ml) after 4th h Mean ±SEM	Percentage protection
1	SAHE	14.66 ± 0.33*	51.61	0.525 ± 0.047*	50.00
2	SCHE	12.00 ± 0.57*	61.29	0.45 ± 0.028*	57.14
3	SRHE	17.66 ± 0.33*	41.94	0.70 ± 0.040*	33.13
4	SSHE	21.33 ± 0.66*	32.26	0.75 ± 0.028*	28.57
5	Control	31.00 ± 0.57	00.00	1.05 ± 0.028	00.00
6	Diclofenac Sodium	07 ± 0.57*	77.14	--	--
7	Paracetamol	--	--	0.35 ± 0.028*	66.67

Values are mean ± SEM (n = 6) * P< 0.001 Vs control; SAHE- Sida acuta Hydro alcoholic extract; SCHE- Sida cordifolia Hydro alcoholic extract; SRHE- Sida rhombifolia Hydro alcoholic extract; SSHE- Sida spinosa Hydro alcoholic extract.

RESULT AND DISCUSSION:

The present investigation revealed that the hydroalcohol extract of roots of Sida acuta, Sida cordifolia, Sida rhombifolia and Sida spinosa were found to be non-toxic even up to a dose as high as 2000 mg / kg body weight. The results are expressed in Table-1.Hydroalcohol extract of four Sida species showed significant analgesic and anti-inflammatory activity. The preliminary phytochemical screening hydroalcohol extract of Sida species showed positive result for alkaloids, flavonoids, tannins, steroids, resins and triterpenoids. Out of four Sida species Sida cordifolia was found to be superior in its analgesic action followed that of Sida acuta and Sida rhombifolia while Sida spinosa was least effective. Sida acuta and Sida cordifolia showed significant reduction in oedema volume after 4th hour at a dose of 200 mg / kg body weight, which is comparable to the standard diclofenac sodium. Sida rhombifolia and Sida spinosa showed mere anti-inflammatory activity in paw oedema method. Further detailed investigations are underway to determine the exact phytoconstituents, which were responsible for the analgesic and anti-inflammatory activity.

REFERENCE:

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8. Winter CA., Risely EA and Nuss GW. Carrageenan-Induced edema in hind paw of the rats as an assay for anti-inflammatory drugs. Exp Bio Med. 1962; 111: 544-547.

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Received on 21.07.2008 Modified on 23.08.2008

Research J. Pharm. and Tech. 1(3): July-Sept. 2008; Page 287-289