Exploration of Anti-inflammatory Activity of Aquaeous Extract of Hibiscus rosa sinesis (Malvacae)

Anshita Gupta¹ , Goutam Kumar Jana², Manmohan Singh Jangdey¹, Deependra Soni²

¹University Institute of Pharmacy, Pt. R.S.U. Raipur -492010 (C. G.).

²Department of Pharmacognosy, Gayatri College of Pharmacy, Jamadarpali, Sambalpur -768200 Orissa.

*Corresponding Author E-mail: anshita1912@gmail.com

ABSTRACT:

In the realm of our Traditional System of Medicine there are a wide range of drugs which have proved their effectiveness by giving a series of excellent performance in acute and chronic diseases. Although the such drugs in concurrent use of conventional system of medicines requires scientific background regarding its therapeutic activity, safety, toxicity, drug interactions etc. One of such drug is Hibiscus rosa sinesis belonging to the family Malvacae. The whole plant exhibit a great chain of activity like antimicrobial, antipyretic, anti-inflammatory, antiimplantation, hair growth accelerator, etc. From the consideration of ethanomedicinal documentations, the aqueous extract of the root part of selected plant is considered for anti-inflammatory activity. The investigatory study of anti-inflammatory activity on the aquaeous root extract of Hibiscus rosa sinesis was carried out using a dose of 250mg/ kg body weight in carrageenan induced paw edema in swiss albino rats .The aquaeous extract produced statistically significant & dose dependant inhibition of edema induced by carrageenan at the dose when compared to the control groups, injected into sub plantar tissue of left paw of animals up to the concentration of 1% w/v. The aquaeous extract of the root (250 mg/kg, orally) gave significant (p<0.01) reduction of rat paw edema at all assessment times.

KEYWORDS: Anti inflammatory, Carrageenan, paw edema, diclofenac sodium.

INTRODUCTION:

Inflammation is a dynamic process that is elicited in response to mechanical injuries, burns, microbial infections, and other noxious stimuli that may threaten the well-being of the host. This process involves changes in blood flow, increased vascular permeability, destruction of tissues via the activation and migration of leucocytes with synthesis of reactive oxygen derivatives (oxidative burst), and the synthesis of local inflammatory mediators, such as prostaglandins (PGs), leukotrienes, and platelet-activating factors induced by phospholipase A2, cyclooxygenases (COXs), and lipoxygenases. Arachidonic acid is a key biological intermediate that is converted in to a large number of eicosanoids with potent biological activities.

The two major pathways of arachidonic acid metabolism are the COX pathway, which results in the formation of both PGs and thromboxanes, and the 5-lipoxygenase pathway, which is responsible for the formation of leukotrienes and 5S-hydroxy-6E,8Z, 11Z, 14Z-eicosatetraenoic acid (5-HETE).[1] Classic examples of herbs traditionally used to treat inflammation in Western medicine are Matricaria chamomilla L. and Arnica montana L. (Asteraceae), Salix alba (Salicaceae), and Glycyrrhiza glabra (Fabaceae). One of such drug is Hibiscus rosa sinesis belonging to the family Malvacae. The whole plant exhibit a great chain of activity like antimicrobial, antipyretic, anti-inflammatory, antiimplantation, hair growth accelerator, etc.

The contamination of herbal material has been documented throughout history but, in recent years another type has occurred. The so-called ‘herbal’ material is mixed with, or replaced by, synthetic pharmaceuticals that show the activity claimed for the herbal. The most common cases are the addition of steroids such as betamethasone to anti-inflammatory ‘herbal’ topical preparations for skin complaints, such as eczema,etc. Herbal extracts are complex mixtures, so high sensitivity and miniaturised assays are required, together with innovative technologies.[2,3]

MATERIAL AND METHODS:

Plant Selection- The root part of Hibiscus rosa sinesis was collected from Sambalpur district Orissa , in the month of April 2010.It was further identified and authenticated by the Botanical Department, Howarah, Kolkata as Hibiscus rosa sinesis belonging to the family Malvacae. Some voucher specimen numbers were submitted to the authority for future references.

Extraction Procedure

The root plant parts were dried in shade and powdered to get a coarse powder. About 2.5kg of dry coarse powder was extracted with Distilled water (40-60⁰C) by hot extraction method. [4] The aquaeous extract was filtered and concentrated to a dry mass by using vacuum distillation. A deep brownish viscous residue obtained having characteristic mucilaginous odour (yield=4.5gm). Further the solvents was evaporated to dryness and then the residue of extract obtained is evaporated to dried mass and then it was taken for the experiment.

Chemicals:

The following drugs and chemicals were used. Drugs: Diclofenac Sodium, (Mankind Pharma Ltd., New Delhi) and all other chemicals were of analytical grade.

Animal-

For performing animal study prior permission was taken from Ethical Committee working at the Institutional level GCOP, Sambalpur, Orissa .Experiments were performed with Swiss Albino rats weighing between 100 to 200gms. The animals were housed in colony cages (three per cage) under conditions of standard lighting, temperature (25±2ºC) and humidity for at least one week before the beginning of experiment, to adjust to the new environment and to overcome stress possibly incurred during transit.[5,6] During this period, they had free access to food and water.

Experimental Assessment of Anti-Inflammatory Activity-

Aquaeous extract of root part of plant was evaluated for anti-inflammatory activity by carrageenan –induced paw edema method in rats .Swiss albino rats (100-200 gm) were randomly distributed into 3 groups of 6 animals each. They were fed with standard diet. The first group served as a control, second group as Standard group was treated with standard anti-inflammatory agent (received Diclofenac sodium 45mg/kg i.p.).Test group was taken for aquaeous extract of root in the concentration of 250mg/kg body wt .After 1 hr,0.1ml of 1%w/v suspension of carrageenan was injected into sub plantar region of the right hind paw to all the 3 groups of rats. The paw volumes were measured using paw immersion method. After carrageenan injection ,mean increase in paw volume were noted for every 1 hrs till 4hrs.

Statistical Analysis of the data

The calculation of the experimental data is done by using One-Way analysis of variance (ANOVA).Later on the calculation is done by various non parametric test. The mean and standard deviation is obtained. The results were presented as Mean ± SEM. Differences between means of treatment and control groups were accepted significant at p<0.001

RESULT AND DISCUSSION:

Effect of Aquaeous root extract of Hibiscus rosa sinesis in reducing Inflammation:

The root extract of Hibiscus rosa sinesis showed significant reduction in paw edema in swiss albino rats when administered at a dose of 250mg/kg body weight in carrageenan induced inflammation. The extract showed slow onset of action but duration of action was longer and also exhibit dose dependent activity. The extract showed significant anti-inflammatory activity. The data obtained in the experiment is shown in Table-1.

CONCLUSION:

The Aqueous Root extract of Hibiscus rosa sinesis in the above experiment was investigated for anti-inflammatory activity at a dose of 250mg/kg body weight. It is reported to contain anthocyanins, flavonoids, cyclopeptide alkaloid, vitamins.etc. The plant Hibiscus rosa sinesis is a gift of nature having a wide range of chemical constituents showing different sets of activities. Although various attempt has been done to explore the maximum therapeutic results but a portion of the plant is still beyond scientific exploration The root part of the plant has immense possibilities of having desired activity profile. The need of the hour is for more experimental studies and scientific data regarding its isolation, characterization molecular assessment of the structure, for the optimization and development of lead compound.

Table 1 Anti-inflammatory Activity of the aqueous extract root of Hibiscus rosa sinesis Linn. in albino rats

S. No	Treatment	Mean Paw Edema in Volume (ml) ± S.E.M
S. No	Treatment	0 MIN	30 MIN	60MIN	120 MIN
1.	Control	2.2±0.185	1.8±0.033	1.47±0.047*	1.54±0.09**
2.	Diclofenac Sodium(45mg/kg)	1.8±0.031	1.76±0.085*	1.32±0.042**	1.23±0.046
3.	Aquaeous Extract(250mg/kg)	2.1±0.14	1.82±0.08	1.50±0.036*	1.19±0.028**

The results are expressed as mean ± S.E.M. The statistical analysis was performed by analysis of variance (A.N.O.V.A) by Dunnett’s Multiple Comparison test. Mean ± SEM, * p<0.05, ** p<0.001

REFERENCES:

[1] Heinz Lüllmann, M.D., Klaus Mohr, M.D., Lutz Hein, M.D.; Color Atlas of Pharmacology; 3rd edition, revised and expanded; Thieme Stuttgart · New York.

[2] Peter J Houghton; Evaluation of Herbal Medicinal Products;Perspectives on quality, safety and efficacy; First published 2009; Published by the Pharmaceutical Press, London,3-14,327.

[3] V. M. Jadhav, R. M. Thorat, V.J. Kadam , N. S. Sathe, Hibiscus rosa sinensis Linn – ‘Rudrapuspa’’ : A Review; Journal of Pharmacy Research ;2(7), 2009,1168-1173

[4] Alka Sawarkar, C.R. Jangde, P.D. Thakre, Ranu Kadoo and Sushma Shelu, Analgesic Activity of Hibiscus rosa sinensis Linn in Rat, Veterinary World, Vol.2(9):353-354

[5] Gupta V, Bansal P , Garg A , Meena AK; Pharmacopoeial Standardization of Hibiscus rosa sinensis Linn., International Journal of Pharmaceutical and Clinical Research; 1(3): 2009;124-126.

[6] Mhaskar, K.S. and Bletter, E., Eds., In; Kirtikar and Basu´s Illustrated Indian Medicinal plants, Sri Satguru Publications, New Delhi, 1989, 462.

Received on 11.05.2014 Modified on 05.06.2014

Res. J. Pharmacognosy & Phytochem. 6(3): July-Sept.2014; Page 109-111