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Antioxidant, Cytotoxic and Analgesic Activities of the Methanolic Fruit Extract of Terminalia Chebula Retz.

*Shammy Sarwar
Department of Pharmacy, Stamford University Bangladesh, 51, Siddeswari Road, Dhaka-1217, Bangladesh

Asif Hasan Malik
Department of Pharmacy, Stamford University Bangladesh, 51, Siddeswari Road, Dhaka-1217, Bangladesh

Muhammad Ashikur Rahman
Department of Pharmacy, Jahangirnagar University, Savar, Dhaka, Bangladesh

Md. Zakiur Rahman
Department of Pharmacy, East West University, Dhaka-1212, Bangladesh

Md. Sohel Rana
Department of Pharmacy, Jahangirnagar University, Savar, Dhaka, Bangladesh

Abstract/ Executive Summary:

The present study was aimed to investigate antioxidant, analgesic and cytotoxic activity of the methanolic extract of Terminalia chebula Retz. fruits. Antioxidant potential of the extract was evaluated by using nitric oxide scavenging assay, reducing power and total antioxidant capacity. The extract showed significant activities in all antioxidant assays compared to ascorbic acid in a dose dependent mode. In nitric oxide scavenging assay, the IC50 value of the extract was found to be 51.3 µg/mL while the IC50 value of ascorbic acid was 77.4 µg/mL. In addition to strong reducing power, total antioxidant activity of the extract was also found to increase in a dose dependent manner. The analgesic activity was evaluated using acetic acid-induced writhing test in mice. The extract, at a dose of 500 mg/kg, showed a maximum of 44.17% inhibition (p < 0.05) of writhing reaction compared to the reference drug diclofenac-sodium (66.96 %). The extract also showed moderate cytotoxic activity in brine shrimp lethality bioassay and the LC50 value was found to be 97.36 µg/mL.

Keywords: Terminalia chebula Retz., Antioxidant, Analgesic, Cytotoxicity.

Location:

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Non-commodity: Medicinal Plants

Objectives:

In this study it was aimed to investigate antioxidant, analgesic and cytotoxic activities of the methalonic extract of T. chebula.

Methodology:

Plant material: The plant Terminalia chebula Retz. was collected from Dhaka in the month of May 2013. Extraction: The powdered plant sample (500 g) was soaked in 1.5 L of methanol for 16 days and then filtered through a cotton plug followed by Whattman filter paper number 1. The extract was concentrated with a rotary evaporator and it afforded 15 g of the methanol extract. Animal: For the experiment Swiss albino mice of either sex, 3-4 weeks of age, weighing between 20-25gm, were collected from the animal research branch of the International Center for Diarrheal Disease and Research, Bangladesh (ICDDR, B). All protocols for animal experiment were approved by the institutional animal research ethical committee. Phytochemical screening: The freshly prepared crude extract was qualitatively tested for the presence of various phytochemical constituents. Phytochemical screening of the extract was performed using the following reagents and chemicals: alkaloids with Dragendorff’s reagent, flavonoids with the use of Mg and HCl; tannins with ferric chloride and potassium dichromate solutions and saponins with ability to produce stable foam and steroids with LibermannBurchard reagent. These were identified by characteristic color changes following standard procedures described by Ghani (2005). Antioxidant Assays: Assay of nitric oxide scavenging activity- The procedure was based on the method, where sodium nitroprusside in aqueous solution at physiological pH spontaneously generates nitric oxide, which interacts with oxygen to produce nitrite ions that can be estimated using Greiss reagent. For the experiment, sodium nitroprusside (10mM) in phosphate buffered saline (PBS) was mixed with different concentrations of methanolic extract of T. chebula dissolved in methanol and incubated at room temperature for 150 minutes. After the incubation period, 0.5 ml of Greiss reagent was added. The absorbance of the chromophore formed was read at 546 nm using a Hach, DR-4000U spectrophotometer. Determination of total antioxidant capacity: The antioxidant activity of the extracts was evaluated by the phosphomolybdenum method according to the procedure of Prieto et al. (1999). The assay is based on the reduction of Mo (VI)-Mo (V) by the extract and subsequent formation of a green phosphate/Mo(V) complex at acid pH. 0.3 ml extract was combined with 3ml of reagent solution. The tubes containing the reaction solution were incubated at 95°C for 90 minutes. Then the absorbance of the solution was measured at 695 nm using a spectrophotometer (Hach, DR-4000U) against blank after cooling to room temperature. Methanol (0.3 ml) in the place of extract is used as the blank. The antioxidant activity is expressed as the number of equivalents of ascorbic acid dissolving required amount of extract in specific volume of pure dimethyl sulfoxide (DMSO). Preparation of test groups: 20 mg of sample was dissolved in 2 ml of DMSO to obtain a solution having concentration of 10 µg/ml. From that test solution different volumes were added to premarked glass vials or test tubes containing 5 ml of seawater and 10 shrimp nauplii. Counting of nauplii: After 24 hours, the vials were inspected using a magnifying glass and the number of survived nauplii in each vial was counted. From this data, the percent (%) of lethality of the brine shrimp nauplii was calculated for each concentration. The median lethal concentration (LC50) of the test samples was obtained by a plot of percentage of the shrimps killed against the logarithm of the sample concentration. Analgesic Screening: Acetic Acid-Induced Writhing Test The analgesic activity of the samples was also studied using acetic acid-induced writhing model in mice. Test samples and vehicle were administered orally 30 min before intraperitoneal administration of 0.7% acetic acid but Diclofenac-Na was administered intraperitonially 15 min before injection of acetic acid. After an interval of 5 min, the mice were observed for specific contraction of body referred to as ‘writhing’ for the next 10 min (Ahmed et al., 2004). Statistical Analysis: Statistical analysis for animal experiment was carried out using one-way ANOVA followed by Dunnet’s multiple comparisons. The results obtained were compared with the vehicle control group. p values < 0.05 were considered to be statistically significant when compared to the control.

Source of Information: International Current Pharmaceutical Journal, December 2013, 3(1): 219-222

Article Link (Online Journal): http://www.banglajol.info/index.php/ICPJ/article/viewFile/17296/12137

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

In conclusion, it can be said that the antioxidant, analgesic and cytotoxic activities shown by the Terminalia chebula Retz. fruit extract lend credence in favor of the various uses of Terminalia chebula Retz. in folk medicine. However, extensive pharmacological studies in molecular level are required to understand underlying mechanism of these actions and eventually to isolate active compounds responsible for such activities in T. chebula fruit extract.

Knowledge Management: Journal