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In Vitro Cytotoxic and Anthelmintic Activities of Leaf and Root Extracts of Leonurus Sibiricus L.

M. R. Saha
Department of Pharmacy, Noakhali Science and Technology University, Sonapur, Noakhali-3814, Bangladesh

F. M. S. N. Ul Bari
Department of Pharmacy, Noakhali Science and Technology University, Sonapur, Noakhali-3814, Bangladesh

M. A. Rahman
Department of Pharmacy, University of Rajshahi, Rajshahi-6205, Bangladesh

M. A. Islam
Department of Pharmacy, University of Rajshahi, Rajshahi-6205, Bangladesh

Abstract/ Executive Summary:

This study investigated the cytotoxic and anthelmintic activities of Leonurus sibiricus L. (commonly known as Raktodrone in Bangladesh) belonging to the family Labiatae. The dried leaves and roots of L. sibiricus were extracted with methanol and fractionated by modified Kupchan method. The crude methanolic extracts as well as its soluble fractions of petroleum ether, ethyl acetate and chloroform were screened for cytotoxic activity using brine shrimp lethality bioassay. They were found to possess significant cytotoxic activities. The LC₅₀ values of crude extract of leaves and its pet-ether, ethyl acetate and chloroform soluble fractions were 1.0, 2.0, 2.11 and1.33 μg/ml, respectively. On the other hand, the LC₅₀ of crude methanolic extract of roots and fractions of pet-ether, ethyl acetate and chloroform were 2.0μg/ml, 2.81 μg/ml, 3.55 μg/ml and 7.58μg/ml, respectively. Vincristine sulphate was used as positive control. The crude methanol extract of leaves and roots also showed very good anthelmintic activities as determined against the earthworms, Pheretima posthuma. The study confirms the moderate anthelmintic and potent cytotoxic activities of leaf and root extracts of L. sibiricus.

Keywords: Leonurus sibiricus; Anthelmintic activities; Brine shrimp cytotoxicity.

Location: Department of Pharmacy, Noakhali Science and Technology University, Sonapur, Noakhali

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Non-commodity: Performance

Objectives:

To examine the cytotoxic activities of roots and leaves extract of L. sibiricus using brine shrimp lethality bioassay.

Methodology:

2.1. Collection and identification of the plant The plant sample of L. sibiricus was collected from the Noakhali and Madaripur district, Bangladesh in February 2011. The plant was identified and a voucher specimen (Accession number DACB: 35900) representing this collection has been deposited in the Bangladesh National Herbarium, Dhaka, for further reference. 2.2. Preparation, extraction and fractionation of plant material. The dried and powdered leaves and roots (500 gm) were extracted with methanol with occasional shaking and stirring for 7 days. The whole extract was filtered and the solvent was evaporated. A semisolid mass of 35 gm for leaves and 30gm for roots were obtained and fractionated with petroleum ether, chloroform and ethyl acetate by the modified Kupchan method [22]. The subsequent evaporation of solvents afforded leaf and root extracts of petroleum ether, chloroform and ethyl acetate fraction. 2.3. Cytotoxicity screening Brine shrimp lethality bioassay is widely used for cytotoxicity assay to isolate the bioactive compounds. Here, simple zoological organism (Artemia salina) was used as a convenient monitor for the screening. The eggs of the brine shrimp were collected from an aquarium shop (Dhaka, Bangladesh) and hatched in artificial seawater (3.8% NaCl solution) for 48 h to mature the nauplii. The cytotoxicity assay was performed on brine shrimp nauplii using Meyer method. The test samples (extract) were prepared by dissolving them in DMSO (not more than 50 μl in 5 ml solution) and sea water (3.8% NaCl) to attain concentrations of 5, 10, 20, 25 and 50 μg/ml. A vial containing 50 μl DMSO diluted to 5 ml was used as a control. Standard Vincristine sulphate was used as positive control. Then matured shrimps were applied to each of all experimental vials and control vial. After 24 h, 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.

2.4. Screening for anthelmintic activity The anthelmintic assay was carried out as per the method of Ajaiyeoba et al with minor modifications. Adult earthworms were used to study the anthelmintic activity due to its anatomical and physiological resemblance with the intestinal roundworm parasites of human being. Because of easy availability earthworm has been widely used for the initial evaluation of anthelmintic compounds in vitro. Adult earthworm (P. posthuma) were collected (3 –5 cm in length and 0.1–0.2 cm in width weighing about 0.8–3.04 g) from moist soil of a road side canal of Noakhali Science and Technology University, Sonapur, Noakhali. All the worms were washed with normal saline to remove fecal matters. The crude methanolic extracts of leaf and root were weighed and dissolved in 10 ml of distilled water to obtain the concentrations of 10, 20, 30, 40 and 50 mg/ml. Albendazole was used as reference standard (10 mg/ml). Earthworms were divided into twelve groups (each containing three) in petridish. The extracts were applied and the paralysis time and death time of the worm were determined. The time of paralysis was noted when no movement of any sort could be observed except when the worms were shaken vigorously. The times of death of the worms were recorded after ascertaining that worms neither moved when shaken vigorously or when dipped in warm water (50°C).

Source of Information: J. Sci. Res. 4 (3), 721-727 (2012)

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The very good wormicidal effect of both leaf and root extracts indicated that the extract can be used as to kill parasitic worms of human.

Knowledge Management: Journal