Evaluation of Analgesic and In-vitro Anti-Inflammatory potential of Fruit Flesh Extract of Terminalia  catappa Linn.

 

R, Ganer*, M, A. Kamble*, D. M. Dhabarde, A. R. Ingole, Dr. J. R. Baheti

Kamla Nehru College of Pharmacy, Butibori, Nagpur-441108 (M.S.) India

*Corresponding Author E-mail: manish.kamble21@gmail.com

 

ABSTRACT:

Terminalia catappa Linn is commonly known as tropical almond or Indian almond. The leaf bark and fruits has been used from a long time as a folk medicine in anti-diarrheic, anti-pyretic and as hemostatic. Leaf has been reported to possesses anti oxidative, Hepatoprotective, antidiabetic, anti inflammatory and anti HIV reverse Transcriptase activity. Fruits of Terminalia catappa contains number of hydrolysable tannins also contain flavonoids. Different concentrations of the aqueous extract of Terminalia catappa fruits flesh extract have been evaluated for their analgesic as well as HRBC membrane stabilization activity. The percent inhibition was found 52.16 (80mg/ml) of test sample which is comparable to the percent inhibition of 64.85 (Table 1)for standard Aspirin (100 mg/ml) Also, high concentration (1000 µg/ml) of the fruit flesh extract was found to stabilize the HRBC membrane up to 65.37% (Table 2), which is comparable to the activity of the standard analgesic Aspirin (76.86%).

 

KEYWORDS: Terminalia catappa, Analgesic, anti-inflammatory, HRBC.

 

 

 


INTRODUCTION:

Terminalia catappa also known as Indian almond or tropical almond is a native throughout the tropical Asia belongs to family Combretaceae. It is widely cultivated and naturalized tree. The tree is often characterized by horizontal branches that are divided repeatedly into tiered whorls. Each tier is usually 3-6 feet apart on the trunk. The leaves are alternate and crowded together near the ends of the twigs in terminal rosettes. Fruits are drupes about the size and shape of an almond fruit but with a slight wing. They are 2 inches or longer and 1 inch across. Full-sized fruits are at first green and turn red, brown, or yellow at maturity.

 

When young, the fleshy fibrous pulp surrounding the large seed is edible and sweet and slightly sour thereafter. Inside the husk there is a light brown, thick, hard stone that contains an almond-like kernel that is also edible.  The leaf bark and fruits has been used from a long time as a folk medicine in anti-diarrheic, anti-pyretic and as hemostatic1. Leaf has been reported to possesses anti oxidative2, Hepatoprotective3, antidiabetic4, anti inflammatory5 and anti HIV reverse Transcriptase activity6. Terminalia catappa contains number of hydrolysable tannins such as pinicalagin, pumcalin, chebulic acid, corialgin, geranin, terflavin7 etc, also contain flavonoids like isovitexin, vitexin, rutin etc8.

 

The presents study being carried out to evaluate analgesic and anti inflammatory activity of fruit flesh extract of Terminalia catappa. Earlier it is reported the same activity with aqueous juice prepared by maceration from tender leaves of Terminalia catappa to support its traditional use9. The therapeutic compound found in the market produces undesirable effects which provided an opportunity to develop new and better compounds. Bioactive compounds derived from plants have been used since long time for various diseases including pain and inflammation as pain and inflammation control is one of the most important therapeutic priorities. Moreover, lot of plants species were identified and possessing analgesic as well as anti inflammatory activity10-11. Inflammation is a response to injury, infection or destruction characterized by, redness, pain, swelling, heat and disturbed physiological function. Inflammation is a normal protective response to tissue injury caused by physical trauma, noxious chemical or microbial agents. It is the body response to inactivate or destroy the invading organisms, to remove the irritants and set the stage for tissue repair. It is triggered by the release of chemical mediators from injured tissue and migrating cells. The commonly used drug for the management of inflammatory conditions are non-steroidal anti-inflammatory drugs, which produce several adverse effects especially gastric irritation leading to formation of gastric ulcers12.

 

MATERIAL AND METHOD:

Collection of Plant Materials:  The fruits of Terminalia cataapa were collected from college campus of Kamla Nehru College of Pharmacy, Butibori, Nagpur Maharastra. Plant material was identified and authenticated in the department of Botany, RTM Nagpur University Nagpur, Maharashtra. The collected materials were cleaned and flesh was removed for further processes of extraction.

 

Preparation of the Extract

Fruit Flesh was taken (100g) and subjected to hydroalcoholic extraction (Soxhlet Extraction) Then, it was filtered and the filtrate was allowed to evaporate under rotary vacuum evaporator until semi solid consistency is obtained. Then the aqueous extract was then re-dissolved in water at 1 mg/ml ratio and used for evaluating in analgesic and in-vitro anti-inflammatory potentials.

 

Analgesic activity:

Formalin induced pain:

In this study a modified method was used13. The pain was induced by injecting 0.05ml of 2.5% formaldehyde (40%) in saline solution through subplantar  region of rat hind paw. Preliminary oral LD50 dose of extract was more than 1000mg/kg hence rats were given with one/10th LD50 dose of extract i.e. 50 and 100mg/kg p.o., aspirine (100mg/kg, p.o.). The time spent in licking the injected paw was considered as indicative and nociceptive pain and was recorded. Two successive phases are involved following the injection of formalin, first phase (0-5min) representing neurogenic pain and second phase (15-30 min) representing panic and inflammatory pain response. The percentage inhibition for each rat and every group was observed as shown in table 1:

 

Percentage inhibition =[ (A0-A1/A0)×100], where A0 is the reading of the co-solvent and A1 is the Sample.

 

Table 1: Effect of T.catappa fruit flesh extract, and Aspirin on Formalin – induced pain

Total Time spent in licking (Sec)

 

Inhibition

Inhibition

Treatment

Dose (mg/Kg)

0-5 min

(%)

15-30 min

(%)

Control

0

55.31± 3.30

 

98.00±

2.61

 

Extract

40

49.56±

1.03

6.12

71.87±

1.69

24.0

Extract

80

45.39±

2.32

18.23

52.16±

3.62

52.16

aspirin

100

52.48±

1.33

5.30

67.82±

2.43

64.85

Each value represent mean ±SEM in seconds for six rats treated with extract or reference drugs.

 

Invitro anti-inflammatory activity:

HRBC Membrane Stabilization

Human red blood cells (HRBC) suspension was prepared according to the previously described method14. The blood was collected from healthy human volunteers who have not taken any NSAIDs for 2 weeks prior to the experiment and transferred to the centrifuge tubes. The tubes were centrifuged at 3000 rpm for 10 min and were washed three times with equal volume of normal saline. The volume of blood was measured and re-constituted as 10% v/v suspension with normal saline. Hypotonicity-induced haemolysis was used for membrane stabilization assay. The reaction mixture (4.5 ml) consisted of 2 ml hypotonic saline (0.25% NaCl), 1 ml of 0.15 M phosphate buffer (pH 7.4), 1 ml extract (200, 400, 600, 800 and 1000 µg/ml) in normal saline and 0.5 ml of 10% human RBC in normal saline. In blank 1 ml of isotonic saline was used instead of extract while control was devoid of red blood cells. The mixtures were incubated at 50oC for 30 min. The tubes were cooled under running tap water for 20 min followed by centrifugation at 1500 rpm for 10 min. Absorbance of the supernatant was read at 560 nm.

 

Membrane stabilization was calculated by using the formula

(Abs of blank - Abs of extract) / Abs of control x 100. The control represents 100%

 

Drugs interacts with membrane hence the model used in this study is Erythrocytes membrane stabilization15-16. NSAIDs stabilize erythrocytes against stress hemolysis. Moreover, they prevent the release of hemoglobin as a result of their membrane stabilizing activity17. The human red blood cells (HRBC) model is selected to assess the anti-inflammatory activity of Terminalia catappa. In this study, 05 different concentrations of the aqueous extract of Terminalia catappa fruits flesh extract have been evaluated for their HRBC membrane stabilization activity. High concentration (1000 µg/ml) of the fruit flesh extract was found to stabilize the HRBC membrane up to 65.37% (Table 2), which is comparable to the activity of the standard analgesic Aspirin (76.86%). All the extracts exhibited membrane stabilization activity in a dose-dependent manner.

 

Table 2: HRBC membrane stabilization activity of the aqueous extract of Terminalia catappa

 Concentration (µg/ml)

Stabilization (%)

62.5

56.32±0.35

125

58.28±0.19

250

61.19±0.77

500

63.80±0.10

1000

65.37±0.26

Aspirin (100µg/ml)

77.86±0.51

 

 

CONCLUSION:

Present study deals with significant analgesic activity and in vitro study on anti-inflammatory potentials of Terminalia catappa fruits flesh. Results obtained from the present study provide scientific evidences for the use of this plant in folk medicine. Further, the present study suggests that Terminalia catappa could serve as a lead in the development of a novel herbal analgesic and anti-inflammatory agent.

 

The study findings showed that the hydro alcoholic extract of Terminalia catappa possess analgesic activity, also the extract showed a significant inhibition of formalin induced hind paw licking in rats. The inhibition is dose dependent with high doses of the extract showing a significance percentage of inhibition. The analgesic and anti-inflammatory effects exhibited by the extracts could as well be attributed to other phytochemicals present in the fruit flesh. Past studies have shown that phytochemicals including tannins and flavonoids possess analgesic and anti-inflammatory activities18-19. To better understand the mechanisms by which phytochemicals of Terminalia catappa  act, further studies are warranted.

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Received on 07.06.2017       Modified on 22.09.2017

Accepted on 20.10.2017       ©A&V Publications All right reserved

Res.  J. Pharmacognosy and Phytochem. 2017; 9(4): 228-230.

DOI: 10.5958/0975-4385.2017.00042.5