Anti Inflammatory activity of methanol extract of Acacia leucophloea Roxb stem bark

About Authors:
Rajesh Gupta¹*, Arun Kumar², Aiswarya G³ , Santanu Roychowdhury^1
1Department of Pharmacognosy,
Sri Sai College of pharmacy, Dalhousie road,
Badhani, Pathankot-145001,
Punjab, India.
²Department of Pharmacy,
BPS Mahila Vishwavidyalaya,
Khanpur Kalan, Sonipat
Haryana,India
³JDT Islam College of Pharmacy,
Vellimadukunnu, Calicut,
Kerala, India

ABSTRACT
The plant Acacia leucophloea Roxb is reported to have great medicinal value in Indian medicine. The present study deals with the preliminary phytochemical screening of Acacia leucophloea Roxb, stem bark in vivo evaluation of anti-inflammatory activity of methanol extract by using Carrageenan induced paw oedema and cotton pellet –induced granuloma model.The cotton pellet granuloma method is widely used to evaluate the transudative and proliferative components of the chronic inflammation. The moist weight of the cotton pellet correlates with the transuda; the dry weight of the pellet correlates with the amount of the granulomatous tissue. Administration of stem bark methanol extract at doses of 100 and 200 mg/kg b.w. and phenylbutazone (100 mg/kg b.w.) appears to be effective in inhibiting the dry weight of the cotton pellet. These data support the hypothesis of the greater effect of the extract on the inflammation mediators in the immediate response of inflammation in rats.

Reference Id: PHARMATUTOR-ART-1214

INTRODUCTION
Acacia leucophloea Roxb also called reonja, is a moderate sized tree and it attains a height of about 20 to 30 ft and a girth of 2 to 3 ft¹, belongs to the family Fabaceae under the subfamily Mimosoideae. Acacia leucophloea’s native range through South and Southeast Asia is non-contiguous². Its largest continuous distribution is arid India through Sri Lanka, Bangladesh, Burma and much of Thailand³. The chemical constituents found are n-Hexacosanol, beta- Amyrin, beta-Sitosterol and Tannin⁴. Traditionally the bark is used as astringent, bitter, thermogenic, styptic, alexeteric, antihelmintic, vulnerary, demulcent, constipating, expectorant and antipyretic, vulnerary, demulcent, constipating, bronchitis, cough, vomiting, wounds, ulcers, diarrhoea, dysentery, internal and external haemorrhages, dental caries, oral ulcers, proctoptosis, stomatitis and intermittent fevers⁵.
Inflammation (Latin, inflammare, to set on fire) is part of the complex biological response of vascular tissues to harmful stimuli, such as pathogens, damaged cells, or irritants. Inflammation is a protective attempt by the organism to remove the injurious stimuli and to initiate the healing process. Inflammation is not a synonym for infection. Anti-inflammatory refers to the property of a substance or treatment that reduces inflammation⁶.The literature survey also revealed that there are no reports on correlation between chemical constituents and their pharmacological properties. The present study is therefore undertaken, to study the Anti-inflammatory activity of the stem bark of Acacia leucophloea.

MATERIAL AND METHOD
Collection of Plant Materials
Dried stem bark of Acacia leucophloea were collected and authenticated by Dr. S.N. Sharma, Technical Officer, Department of plant Sciences, Indian Institute of Integrative medicine, Jammu. A voucher specimen (specimen No. 21852) was deposited in the herbarium of Indian Institute of Integrative medicine, Jammu. All the animal experiments were conducted by the approval of Institutional Animal Ethics Committee (reg. no- 1349/AC/10/CPCSEA). During the study period, guidelines of Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Institutional Animal Ethics Committee (IAEC) were followed for the maintenance of animals.  

 Methods
a. Method of preparation of methanol extract
The extraction was done by using hot continuous method using Soxhlet apparatus. The   successive extraction was performed by using the solvents n-hexane and methanol. The methanol extract was used for this study.

b. Anti-inflammatory activity by using
i) Carrageenan induced paw oedema⁷
The animals were divided into four groups with six animals in each group. The animals were adminstered simultaneously with extract or phenylbutazone with 0.1 ml of 1% carrageenan in normal saline in the sub-plantar region of right hind paw. Paw volume was measured by Plethysmometerically at 0and 3 h after carrageenin injection.  Group I animals served as control received normal saline 3ml/kg body weight (orally), animals of Group II received phenylbutazone at 100 mg/kg body weight (orally) while animals of Group III and Group IV were treated with 100 and 200 mg/kg body weight (orally) of the methanol extract of stem bark. Reduction in the paw volume compared to the vehicle-treated control animals was considered as anti-inflammatory response. Mean increase in paw volume was measured and percentage of inhibition was calculated.
ii) Cotton pellet-induced granuloma⁸
Subacute inflammation was produced by cotton pellet induced granuloma in rats. Sterile cotton (50 ±1 mg) soaked in 0.2 ml of distilled water containing penicillin (0.1 mg) and streptomycin (0.13 mg) was implanted subcutaneously bilaterally in axilla under ether anaesthesia. Group I animals served as control received normal saline 3ml/kg body weight (orally), animals of Group II received phenylbutazone at 100 mg/kg body weight (orally) while animals of Group III and Group IV were treated with 100 and 200 mg/kg body weight (orally) of the methanol extract of stem bark for consecutive six days. The animals were sacrificed on the 7th day. The granulation tissue with cotton pellet was dried at 60^oC overnight and then the dry weight was taken.

RESULT
The anti-inflammatory activity of stem bark methanol extract was measured   at the dose of 100 and 200mg/kg b.w. against carrageenin-induced paw oedema and cotton pellet-induced granuloma. The methanol extract produced significant (P < 0.05) anti-inflammatory   activity and the results were comparable to that of phenylbutazone as a standard anti-inflammatory drug. The stem bark extract at doses 200 mg/kg showed an inhibition of 52.54% (Table no. 1) against paw oedema induced by carrageenin, with comparable to standard drug (62.71%).  The stem bark extract at doses 200 mg/kg showed an inhibition of 55.42% against cotton pellet-induced granuloma, with comparable to standard drug (57.53%). There was dose dependant reduction in granular tissue formation in extract and phenylbutazone treated rats as shown in Table 1.

DISCUSSION
Histamine is one of the important inflammation mediators and it is a potent vasodilator substance and increases the vascular permeability. This study showed that all the doses of stem bark and barks extracts effectively suppressed the oedema produced by histamine, so it may be suggested that its anti-inflammatory activity is possibly backed by its antihistaminic activity. The extracts may exhibit its anti-inflammatory action by means of inhibiting the synthesis, release or action of inflammatory mediator’s viz. histamine that might be involved in inflammation. From the above results it is suggested that the anti-oedematogenic effects of both extracts on carrageenin-induced paw oedema may be related to inhibition of inflammation mediator formation.The cotton pellet granuloma method is widely used to evaluate the transudative and proliferative components of the chronic inflammation. The moist weight of the cotton pellet correlates with the transuda; the dry weight of the pellet correlates with the amount of the granulomatous tissue. Administration of stem bark extract at doses of 100 and 200 mg/kg b.w. and phenylbutazone (100 mg/kg b.w.) appear to be effective in inhibiting the dry weight of the cotton pellet. These data support the hypothesis of the greater effect of the extract on the inflammation mediators in the immediate response of inflammation in rats. This effect may be due to the cellular migration to injured sites and accumulation of collagen and mucopolysaccharide.

CONCLUSION
The present study suggest that the plant Acacia leucophloea Roxb could be a potential source as natural anti-inflammatory that could have great importance to inhibition of inflammation mediator formation.

REFERENCES
1. Kaul RN. Need for afforestation in the acrid zone of India. 13.India: Layaaran; 1963.
2. en.wikipedia.org/wiki/ Acacia leucophloea.
3. Nielsen IC. Flora Malesiana: Mimosaceae.1. France: Kluwer; 1992.
4. Department of Indian System of Medicine and Homoeopathy. The Ayurvedic            Pharmacopoeia of India. I (2). India: The Controller of Publications; 1999.
5. Bhadoria BK, Gupta RK. A note on hydrocynic  acid content in Acacia leucophloea   Roxb Willd. Current  Science 1981; 50: 689-90.
6.  en.wikipedia.org/wiki/Inflammation.
7. Winter CA, Risley EA, Nuss GW. Carrageenin-induced edema in hind paw   of the    rat as an assay for anti-inflammatory drug. In Proceedings of the Society for Experimental Biology and Medicine 1962; 11: 544–547.
8. Swingle KF, Shideman FE. Phases of the inflammatory response to subutaneous implantation of a cotton pellet and their modification by certain anti-inflammatory agents. Journal of Pharmacology and Experimental Therapeutics 1972;183: 226–234.

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