EVALUATION OF THROMBOLYTIC AND MEMBRANE STABILIZING ACTIVITIES OF FOUR MEDICINAL PLANTS OF BANGLADESH

EVALUATION OF THROMBOLYTIC AND MEMBRANE STABILIZING ACTIVITIES OF FOUR MEDICINAL PLANTS OF BANGLADESH

S.R. Chowdhury, T. Sharmin*, M. Hoque, Md. Sumsujjaman, M. Das and F. Nahar

Department of Pharmacy, State University of Bangladesh, Dhaka, Bangladesh

ABSTRACT: The crude methanol extracts of aerial parts ofAbrus precatorius L., leaf of Magnolia pterocarpa Roxb. and Dracaena spicata Roxb. and leaf and bark ofRavenala madagascariensis Sonn. as well as their hexane, carbon tetrachloride, chloroform and aqueous soluble partitionates were subjected to screenings for thrombolytic and membrane stabilizing activities. Among the extractives of A. precatorius, the crude methanol extract exhibited the highest thrombolytic activity (34.92±0.54 %) while the carbon tetrachloride soluble fraction of M. pterocarpa exhibited 22.59±0.88 % clot lysis. D. spicata extractives showed mild thrombolytic activity. The methanolic crude extract of R. madagascariensis leaf and the aqueous soluble fraction of R. madagascariensis bark extract showed 45.32±0.82 % and 32.67±0.74% clot lysis, respectively. In hypotonic solution and heat induced conditions, the crude methanol extract of A. precatoriusand the hexane soluble fraction of crude methanol extract of M. pterocarpa inhibited 63.46±0.84 % & 36.54±0.21 % and 66.12±0.66 % & 40.54±0.02 % haemolysis of RBCs, respectively as compared to 71.90 % and 42.12 % inhibition by acetyl salicylic acid (0.10 mg/ml), respectively. The crude methanol extract of D. spicata demonstrated 64.44±0.68 % and 36.52±0.19 % inhibition of hypotonic solution and heat induced hemolysis, respectively. The chloroform soluble fraction of R. madagascariensis leaf extract demonstrated 28.72±0.61 % & 39.97±0.39 % and the hexane soluble fraction of R. madagascariensis bark extract revealed 53.78±0.17 % & 41.83±0.61 % inhibition of hypotonic solution and heat induced hemolysis of RBCs,  respectively.

Abrus precatorius L., Magnolia pterocarpa Roxb., Dracaena spicata Roxb., Ravenala madagascariensis Sonn., thrombolysis, membrane stabilization

INTRODUCTION:According to the World Health Organization (WHO), 80% of the world’s populations rely on traditional medicines.¹ The practice of herbal medicine is common in rural areas where western medicines are too expensive or not available.¹

Humans have frequently used plants to treat common infectious diseases and some of these traditional medicines are still part of the habitual treatment of various maladies.

It has been reported that 115 articles were published on the antimicrobial activity of medicinal plants in Pubmed during the period between 1966–1994, but in the following decade, between 1995 and 2004, 307 were published ². The demand for more and more drugs from plant sources is continuously increasing. It is therefore essential for systematic evaluation of plants used in traditional medicine for various ailments.

Hence, there is need to screen medicinal plants for promising biological activity ³. Drugs derived from unmodified natural products or drugs semi-synthetically obtained from natural sources corresponded to 78% of the new drugs approved by the FDA between 1983 and 1994 ⁴.

Abrus precatorius L. (Synonyms: Abrus abrus, Glycine abrus; Bengali name: Kunch, Ratii) commonly known as crab's eye, john crow bead, precatory bean and jumbie bea, is a slender, perennial climber of Fabaceae family. The plant is native to India and other tropical and subtropical areas of the world. A tea is made from the leaves and used to treat fevers, cough and cold. Seeds are poisonous and therefore are used after mitigation. The plant is also used in Ayurveda.⁵ An ethanolic extract of seeds of A. precatorius was found to have antioxidant, anti-inflammatory and analgesic potentials in rodents ⁶.

A. precatorius seed extract also caused reversible alterations in the estrous cycle pattern and completely blocked ovulation in Sprague-Dawley rats ⁷. A methanolic extract of the plant produced dose-dependent bronchodilator activity in a guinea pig model ⁸.

Magnolia pterocarpa Roxb. (Synonyms: Lirianthe grandiflora, Liriodendron grandiflorum Roxb., L. indicum Spreng.) locally known as dulichapa, is a flowering medium to large tree of Magnoliaceae family. The plant is native to 3600 feet altitude forests of India, Burma and Pakistan. The bark contains sesamin, eudesmin, fargesin, imperatorin, dimethyl teraphthalate and β-sitosterol. Powdered bark is used for fever and cough ⁹.

Dracaena spicata Roxb. (Synonyms: D. wallichii Kunth., Draco spicata Roxb. Kuntze; Bengali name: ognikundo), commonly known as dragon tree, is a tree of Asparagaceae family. The plant is distributed in Assam, Bangladesh, Andaman Islands and Myanmar. The leaf extract is used by the chakma communities in the treatment of measles ¹⁰.

Leaf juice is used to cure long term fever, coughs and mucus in nose by traditional healers of the Marma tribe of Naikhongchhari, Bandarban District ¹¹.

Ravenalamadagascariensis Sonn. (Synonyms: ­­­­­­­Heliconia ravenala Willemet, Urania madagas- cariensis Sonn. Raeusch; Bengali name: Panthapadak) commonly known as Traveler's Tree or Traveler’s Palm, is a species from Madagascar. It is not a true palm but a member of the bird of paradise family, Strelitziaceae. It is endemic to secondary forests in Madagascar. The leaves have been reported to have anti-diabetic activity in alloxan induced diabetic rats ¹².

As part of our ongoing investigations on medicinal plants of Bangladesh ^{13, 14, 15, 16, 17, 18}, the crude methanol extracts of aerial parts ofA. precatorius, leaf of M. pterocarpa and D. spicata and leaf and bark ofR. madagascariensis growing in Bangladesh, as well as their organic and aqueous soluble fractions were studied for thrombolytic and membrane stabilizing activities for the first time and we, here in, report the results of our preliminary investigations.

MATERIALS AND METHODS:

Collection of plant materials and extraction:The aerial parts ofA. precatorius, leaf of M. pterocarpa and D. spicata and leaf and bark ofR. madagascariensis were collected in March 2012 from Dhaka. Voucher specimens DUSH-10775, DUSH-10774 and DUSH-10777 for collection of A. precatorius, M. pterocarpa and D. spicata have been deposited in Salar Khan Herbarium, Department of Botany, University of Dhaka, respectively. In Bangladesh National Herbarium, voucher specimen DACB 38302 has been deposited for the collection of leaf and bark ofR. madagascariensis. The experiments were conducted in Phytochemical Research Laboratory, State University of Bangladesh in 2012.

The collected plant materials were cleaned, sun dried and pulverized. The powdered materials (500 g each) of the collected plants were separately soaked in 2.0 liters of methanol at room temperature for seven days. The extracts were then filtered through fresh cotton bed and finally with Whatman filter paper number 1 and concentrated with a rotary evaporator at reduced temperature and pressure. An aliquot (5 g) of each of the concentrated methanol extract was fractionated by the modified Kupchan partition protocol ¹⁹ and the resultant partitionates were evaporated to dryness with rotary evaporator to yield hexane (HXSF), carbon tetrachloride (CTCSF), chloroform (CSF) and aqueous (AQSF) soluble materials (Table 1). The residues were then stored in a refrigerator until further use.

TABLE 1: KUPCHAN PARTITIONING OF A. PRECATORIUS, M. PTEROCARPA, D. SPICATA AND R. MADAGASCARIENSIS

ME= Methanolic crude extract; HXSF= Hexane soluble fraction; CTCSF= Carbon tetrachloride soluble fraction; CSF= Chloroform soluble fraction; AQSF= Aqueous soluble fraction

Thrombolytic activity:The thrombolytic activity was evaluated by the method developed by Prasad et al (2006) ²⁰ by using streptokinase as positive control.

Membrane stabilizing activity:The membrane stabilizing activity of the extractives was assessed by evaluating their ability to inhibit hypotonic solution and heat induced haemolysis of human erythrocytes following the method developed by Omale et al (2008) ²¹.

Statistical analysis: For all bioassays, three replicates of each sample were used for statistical analysis and the values are reported as mean ± SD.

RESULTS AND DISCUSSION: The crude methanol extracts of aerial parts ofA. precatorius, leaf of M. pterocarpa and D. spicata and leaf and bark ofR. madagascariensis as well as their hexane, carbon tetrachloride, chloroform and aqueous soluble partitionates were subjected to screenings for thrombolytic and membrane stabilizing potentials. In order to identify the drugs with the ability to promote lysis of blood clot from natural resources, the extractives of A. precatorius, M. pterocarpa, D. spicata and R. madagascariensis were assessed for thrombolytic activity. Addition of 100 μl streptokinase, a positive control (30,000 I.U.) to the clots of human blood and subsequent incubation for 90 minutes at 37°C showed 66.77 % lysis of clot. On the other hand, distilled water, treated as negative control, revealed a negligible lysis of clot (3.79 %).

Among the extractives of A. precatorius and M. pterocarpa,the crude methanol extract and the carbon tetrachloride soluble fraction exhibited 34.92±0.54 % and 22.59±0.88 % clot lysis, respectively. On the other hand, D. spicata extractives showed mild thrombolytic activity and the highest thrombolytic activity was demonstrated by the carbon tetrachloride soluble fraction (21.05±0.23 %). Different extractives of R. madagascariensis leaf and bark demonstrated mild to moderated clot lysis activity ranging from22.55 % to 45.32 % and 24.44 % to 32.67 %, respectively. The methanolic crude extract of leaf and the aqueous soluble fraction of the bark extract showed 45.32±0.82 % and 32.67±0.74 % clot lysis, respectively (Table 2).

TABLE 2: THROMBOLYTIC ACTIVITIES OF A. PRECATORIUS, M. PTEROCARPA, D. SPICATA AND R. MADAGASCARIENSIS LEAF AND BARK EXTRACTIVES

ME = Methanol crude extract; HXSF = Hexane soluble fraction; CTCSF = Carbon tetrachloride soluble fraction; CSF = Chloroform soluble fraction; AQSF = Aqueous soluble fraction; SK =Streptokinase.

The membrane stabilizing activity of A. precatorius, M. pterocarpa, D. spicata and R. madagascariensis extractives was also determined. In hypotonic solution and heat induced conditions, the crude methanol extract of A. precatoriusinhibited 63.46±0.84 % and 36.54±0.21 % haemolysis of RBCs, respectively as compared to 71.90 % and 42.12 % inhibition by acetyl salicylic acid (0.10 mg/ml), respectively. The hexane soluble fraction of crude methanol extract of M. pterocarpa and the crude methanol extract of D. spicata demonstrated 66.12±0.66 % & 40.54±0.02 % and 64.44±0.68 % & 36.52±0.19 % inhibition of hypotonic solution and heat induced hemolysis, respectively.

The chloroform soluble fraction of R. madagascariensis leaf extract demonstrated 28.72±0.61 % & 39.97±0.39 % and the hexane soluble fraction of R. madagascariensis bark extract revealed 53.78±0.17 % & 41.83±0.61 % inhibition of hypotonic solution and heat induced hemolysis of RBCs,  respectively (Table 3).

TABLE 3: MEMBRANE STABILIZING ACTIVITIES OF A. PRECATORIUS, M. PTEROCARPA, D. SPICATA AND R. MADAGASCARIENSIS LEAF AND BARK EXTRACTIVES

ME= Methanolic crude extract; HXSF= Hexane soluble fraction; CTCSF= Carbon tetrachloride soluble fraction; CSF= Chloroform soluble fraction; AQSF= Aqueous soluble fraction

CONCLUSION: The objective of the study was to evaluate the thrombolytic and membrane stabilizing potentials of crude methanol extracts of aerial parts ofA. precatorius, leaf of M. pterocarpa and D. spicata and leaf and bark ofR. madagascariensis as well as their hexane, carbon tetrachloride, chloroform and aqueous soluble partitionates. It is clearly evident from the above findings that the extractives of A. precatorius, M. pterocarpa, D. spicata and R. madagascariensis bark exhibited mild to moderate thrombolytic activity but the R. madagascariensis leaf extractives demonstratd significant thrombolytic activity.

On the other hand, the extractives of A. precatorius, M. pterocarpa, D. spicata and R. madagascariensis bark exhibited significant membrane stabilizing activity but the R. madagascariensis leaf extractives demonstrated mild to moderate membrane stabilizing activity. Therefore, these plants are good candidates for further systematic, chemical and biological studies to isolate the active principles.

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    How to cite this article:

    Chowdhury SR, Sharmin T, Hoque M, Md. Sumsujjaman, Das M and Nahar F: Evaluation of Thrombolytic and membrane stabilizing activities of four medicinal plants of Bangladesh. Int J Pharm Sci Res 2013; 4(11): 4223-27. doi: 10.13040/IJPSR. 0975-8232.4(11).4223-27

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