International Journal of Drug Development and Research

Key words

Basella alba, anti-inflammatory, Human red blood cell membrane stabilization method

Introduction

Basella alba L., (Basellaceae) commonly has known as “Poi (Hindi), Potaki (Sanskrit) and Pasalakkirai (Tamil) [1]. In present study Diclofenac sodium, a nonsteroidal antiinflammatory drug (NSAID) was is used as reference standard drug and leaf extracts of Basella alba as a test drug. Inflammation is a normal protective response to tissue injury and it involves a complex array of enzyme activation mediator release, fluid extravasations, cell migration, tissue breakdown and repair[2]. which are aimed at host defense and usually activated in most disease condition. The critical role of inappropriate inflammation is becoming accepted in many diseases that affect man, including cardiovascular diseases, inflammatory and autoimmune disorders, neurodegenerative conditions, infection and cancer [3].

In appreciating the inflammatory process, it is important to understand the role of chemical mediators. These are the substances that tend to direct the inflammatory response. These inflammatory mediators come from plasma proteins or cells including mast cells, platelets, neutrophils and monocytes/macrophages. They are triggered by bacterial products or host proteins. Chemical mediators bind to specific receptors on target cells and can increase vascular permeability and neutrophil chemotaxis, stimulate smooth muscle contraction, have direct enzymatic activity, induce pain or mediate oxidative damage. Most mediators are short-lived but cause harmful effects. Examples of chemical mediators include vasoactive amines (histamine, serotonin), arachadonic acids (prostaglandins, leukotrienes) and cytokines (tumor necrosis factor and interleukin–1) [4].

Basella alba L.also known Indian spinach. The aerial part (leaves, stem) of the plant serve as edible plant (vegetable) in many parts of world [5]. These contain different components which have extensively used in constipation, as diuretic, in urticaria, as demulcent, antiulcers, and as cooling application for burn[6]. India, due to its geographical and environmental positioning has traditionally been a good source for such products among the Asian countries.

Material and Methods

Plant Material

Basella alba Linn var. alba leaves for the proposed study were collected from vegetable market of Tripura (India) and were authenticated (Regd.- PARC/2009/258) by Prof. P. Jayaraman, Director, PARC, National Institute of Herbal Sciences, W.Tambaram, Chennai (Tamilnadu).The fresh collected leaves of Basella alba Linn. Were shade dried and used for this study. The coarse powder was subjected to extracted separately using ethanol and water as solvents. The solvents were distilled under reduced pressure using rotary vacuum evaporator. The extracts were dissolved in distilled water.

Phytochemical screening

The Phytochemical examination of the both extracts was performed by the standard methods [11] and shows the presence of various phytochemical constituents tabulated in Table-2.

Method:-

In-vitro anti-inflammatory activity

The human red blood cell (HRBC) membrane stabilization method: The blood was collected from healthy human volunteer who had not taken any NSAIDS for 2 weeks prior to the experiment and mixed with equal volume of Alsever solution(2% dextrose, 0.8% sodium citrate, 0.5% citric acid and 0.42% NaCl) and centrifuged at 3,000 rpm. The packed cells were washed with isosaline and a 10% suspension was made. Various concentrations of extracts were prepared (200 and 400 μg/ml) using distilled water and to each concentration 1 ml of phosphate buffer, 2 ml hyposaline and 0.5 ml of HRBC suspension were added. It was incubated at 370C for 30 min and centrifuged at 3,000 rpm for 20 min. and the hemoglobin content of the supernatant solution was estimated spectrophotometrically at 560 nm. Diclofenac (100 and 200 @g/ml) was used as reference standard and a control was prepared by omitting the extracts. [7].

The percentage of HRBC membrane stabilization or protection was calculated by using the following Formula,

Results and Discussion

The results are reported in table 1. The petroleum ether, chloroform, ethyl acetate, methanol and aqueous extracts of the leaves of Basella alba L.were studied for in vitro antiinflammatory activity by HRBC membrane stabilization method. The in vitro anti inflammatory activity of the extracts were concentration dependent, with the increasing concentration, the activity is also increased. Among both the extracts, aqueous extract at a concentration of 400 @g/ml showed 71.89 % protection of HRBC in hypotonic solution. All the results were compared with standard Diclofenac which showed 83.54 % protection.

Basella alba L. leaves extracts exhibited membrane stabilization effect by inhibiting hypotonicity induced lysis of erythrocyte membrane. The erythrocyte membrane is analogous to the lysosomal membrane [8] and its stabilization implies that the extract may as well stabilize lysosomal membranes. Stabilization of lysosomal membrane is important in limiting the inflammatory response by preventing the release of lysosomal constituents of activated neutrophil such as bactericidal enzymes and proteases, which cause further tissue inflammation and damage upon extra cellular release [9] Some of the NSAIDs are known to posses membrane stabilization properties which may contribute to the potency of their anti-inflammatory effect. Though the exact mechanism of the membrane stabilization by the extract is not known yet; hypotonicity-induced hemolysis may arise from shrinkage of the cells due to osmotic loss of intracellular electrolyte and fluid components. The extract may inhibit the processes, which may stimulate or enhance the efflux of these intracellular components. [10] Aqueous extract showed significant in-vitro anti-inflammatory activity as compared to standard. The aqueous extract showed significant anti-inflammatory activity (71.89 %) at the dose of 400 μg/ml. On the basis of the above results it can be concluded that the Basella alba L. have an antiinflammatory activity.

CONCLUSION

Basella leaves extracts exhibited membrane stabilization effect by inhibiting hypotonicity induced lysis of erythrocyte membrane. The erythrocyte membrane is analogues to the lysosomal membrane [8] and its stabilization implies that the extract may as well stabilize lysosomal membrane. Stabilization of lysosomal membrane is important in limiting the inflammatory response by preventing the release of lysosomal constituents of activated neutrophil such as bacterial enzymes and proteases which cause further tissue inflammation and damage[9] From the above study it was concluded that the aqueous and methanolic extracts of leaves of Basella alba L. has significant membrane stabilization property comparable to the standard drug diclofenac.

Acknowledgment

We would like to acknowledge that this research was supported by the Principal Dr.K.S.Laxmi and Dr. K.S.Illango Vice-principal, SRM College of Pharmacy. SRM University, Kattankulathur, Chennai.

Conflict of Interest

NIL

Source of Support

NONE

Tables at a glance

Table 1 Table 2

References

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