Phytochemical Screening and Effect of Musa paradisiaca Stem Extrude on Rat Haematological Parameters

This study was carried out to investigate the phytochemical composition and effect of various concentrations of Musa paradisiaca stem extrude on haematological parameters in Albino Wistar rats. Twenty rats (62-121 g) were randomly assigned into 5 groups of 4 rats each. Group 1 the control group, were given ordinary water while the test groups 2, 3, 4 and 5 were given 25, 50, 75 and 100% of the aqueous extract, respectively without water for 28 days. Rats were sacrificed and blood samples were collected by cardiac puncture then used for the haematological studies. Phytochemical tests were carried out using standard laboratory techniques. The results of the study show the presence of tannins and glycosides in abundance in the stem extrude while saponins, flavonoids, alkaloids, polyphenols and reducing sugars were present in moderate amounts but phlobatannins was absent. There was a significant (p>0.05) increase in rat RBC, PCV, Hb and WBC counts at concentrations of 75 and 100% when compared with the control and a significant (p>0.05) decrease in MCH and MCHC. The levels of MCV were not significantly altered at all extract concentrations. It can therefore be concluded that Musa paradisiaca stem extrude has a haematopoietic and immunomodulatory effect consistent with its ethnomedicinal use.


INTRODUCTION
Musa paradisiaca (family-Musaceae), also known as plantain is a tropical plant that is native to India.It is extensively cultivated in the tropics and is a staple crop for over 70 million people of the sub-Saharan Africa.It is usually cultivated for its carbohydrate content and can be consumed as an unripe fruit or when ripe (Ahenkora et al., 1997).Each stem normally produces a single, sterile, male banana flower, also known as the banana heart.The female flowers appear further up the stem and produce the actual fruit without fertilization.Bananas are one of the most popular fruits on the world market.It is well known that fruits like banana contain various antioxidants, such as vitamin C, vitamin E and β-carotene (Kanazawa and Sakakibara, 2000).Various parts of the plant such as the leaves, roots and flowers have been used for medicinal purposes.For example, the fruit is consumed as food.Again, the leaf juice is used in the treatment of fresh wounds, cuts and insect bites while the leaves act as an arbortifacient.The sap of the plant is used as a remedy for diarrhoea, dysentery, hysteria and epilepsy.A cold infusion of the root is used to treat venereal diseases and anaemia.In addition, the fruit has been reportedly used as antiscorbutic, aphrodisiac and diuretic (Salawu et al., 2010).Along with other fruits and vegetables, consumption of bananas is associated with a reduced risk of colorectal cancer (Deneo-Pellegrini et al., 1996), renal cell carcinoma (Rashidkhani et al., 2005) and breast cancer in women (Zhang et al., 2009).Banana stem extract from the Musaceae family had been suggested to be a useful agent in the treatment of patients with hyperoxaluric urolithiasis (Poonguzhali and Chegu, 1994), kidney stones and high blood pressure.Oral administration of chloroform extract of the Musa sapientum flowers had been found to cause a significant reduction in blood glucose and glycosylated haemoglobin; increase in total haemoglobin and prevents decrease in body weight (Pari and Uma-Maheswari, 1999).This study was designed to evaluate the effects of Musa paradisiaca Stem Extrude on haematological indices in rats.

MATERIALS AND METHODS
Experimental animals: Albino Wistar rats (weighing between 62-121 g) were used for this study.They were obtained from the animal house of National Veterinary Research Institute (NVRI) Vom, Jos and housed at room temperature for 14 days before the commencement of the experiment.
Experimental plant: Matured banana stem was obtained from Professor P.C Onyenekwe garden in Abuja, the capital city of Nigeria.
Preparation of plant extract: The banana stem was washed free of debris.It was sliced and mashed with mortar and pestle to press out the juice.One hundred mL of the pure extract was used to make 100% (v/v) while 75% (v/v) was made by measuring 75 mL of the extract and made up to 100 mL with distilled water in a volumetric flask, 50% (v/v) was made with 50 mL of the extract made up to 100 mL with water while 25% (v/v) was made with 25 mL of the extract made up to 100 mL with water in a volumetric flask.
Experimental protocol: Twenty albino Wistar rats were randomly assigned into 5 groups of 4 rats each.Group 1 the control group was given ordinary water while groups 2, 3, 4 and 5 were the test groups were given 25, 50, 75 and 100%, respectively of the aqueous extract respectively without water for 28 days (4 weeks).All the animals were fed on normal rat chow.The weight of the rats in each group was determined and documented weekly.
Collection of blood samples: Rats were sacrificed using chloroform anaesthesia.Blood samples were collected by cardiac puncture into EDTA capped bottles with the aid of a 2 mL syringe.The blood samples were then used for the experiments.

Phytochemical screening of the plant extract:
The different phytochemical tests were carried out using standard laboratory techniques.Alkaloids, glycosides (Salkowski test) and Saponins (Frothing test) were identified with the method of Sofowora (1984).The method of Trease and Evans (1986) was adopted to test for the presence of anthraquinones, phlobatannins, flavonoids and tannins.

RESULTS AND DISCUSSION
The result of phytochemical screening of waterstem extract of Musa paradisiaca is shown in Table 1.The results show that the stem extrudes of Musa paradisiaca contains tannins and glycosides in abundance.
Alkaloids, saponins, flavonoids, polyphenols and reducing sugars were present in moderate quantities while phlobatannins were absent.Phytochemicals are known to occur in various parts of plants with diverse functions which include provision of strength to plants, attraction of insects for pollination and feeding, defence against predators, provision of colour, while some are simply waste products (Ibegbulem et al., 2003).When ingested by animals these secondary metabolites exhibit varied biochemical and pharmacological actions (Amadi et al., 2006).The presence of tannins in diets for livestock have been reported to have anti-nutritional and toxic effects including reduced fed intake, growth, feed efficiency and net metabolizable energy (Acamovic and Brooker, 2005).However, the results in this research showed that tannins did not impair the growth performance of the albino Wister rats (Table 2).A general increase in physical activities, food and water intake were observed for all the animals during the feeding experiment.The increased weight could be due to increased feed intake observed all through the experimental period which suggests that the rats accumulated calories from the normal rat diet.
Tannins in plants have been shown to confer antidiarrhoeic and anti-haemorrhagic properties on plants (Asquith and Butler, 1986).This is consistent with the traditional use of the sap of Musa paradisiaca for the treatment of diarrhoea, fresh wounds, cuts and insect bites.Saponins have been reported to have antifungal properties (Osuagwu et al., 2007) as well as serve as an expectorant and emulsifying agent (Edeoga et al., 2003).Alkaloids, flavonoids and tannins have been known to show medicinal activity as well as exhibiting physiological activity (Sofowora, 1993).Flavonoids are known to have antioxidant effects and have been shown to inhibit the initiation, promotion and progression of tumors (Kim et al., 1994).The presence of these phytochemicals in the sap of Musa paradisiaca confers medicinal properties on the plant and this explains the use of this plant for treatment of different ailments.The findings of this study is consistent with reports of the presence of these phytochemicals in various parts of the Musa paradisiaca plant as documented by Akpuaka and Ezem (2011) and Akpabio et al. (2012).The assessment of haematological parameters could be used to reveal the deleterious effect of foreign compounds including plant extracts on the blood constituents of animals.They can also be used to determine possible alterations in the levels of biomolecules, metabolic products, haematology, normal functioning and histomorphology of the organs (Magalhaes et al., 2008).Following the administration of the extract, there is a significant (p>0.05)increase in the level of RBC, PCV and Hb and a significant decrease in the levels of MCH and MCHC at extract concentrations of 50-100% while there is no significant change in the level of MCV at all concentrations of the extract when compared with the control as shown in Table 3.
This gives an indication that the plant extract contains phytochemicals that stimulate the formation or secretion of erythropoietin in the stem cells of the animals.Erythropoietin is a glycoprotein hormone which stimulates stem cells in the bone marrow to produce red blood cells (Ohlsson and Aher, 2009).A similar trend was observed when garlic (Allum sativum) extract was administered to rats as reported by Iranloye (2002).Since MCHC, MCH and MCV relate to individual red blood cells while Hb, RBC and PCV relate to the total population of red blood cells in the blood (Adebayo et al., 2005), it could thus imply that though the extract may stimulate the production of red blood cells and haemoglobin, it could have an inhibitory effect on haemoglobin incorporation into red blood cells and a consequent reduction in oxygen exchange.WBC levels increased significantly at extract concentrations of 75 and 100% which shows that the extract of Musa paradisiaca has an immune boosting property similar to that reported for garlic (Allum sativum) by Iranloye (2002) and seed extract of Citrus paradisi Macfad (Adeneye, 2008).It has been reported in literature (Guyton and Hall, 2000;Ganong, 2001) that granulocyte-macrophage colony stimulating factor, macrophage colony stimulating factor, interleukins IL-2, IL-4 and IL-5 regulate the proliferation, differentiation and maturation of committed stem cells responsible for the production of white blood cells.Since the extract of Musa paradisica increases WBC concentration, it may be that some components of the extract stimulated the production of these regulatory factors or increased the sensitivity of the committed stem cells (responsible for the production of white blood cells) to these factors.

CONCLUSION
It can be concluded from this study that the stem extrude of Musa paradisiaca contains phytochemicals which confer medicinal properties on the plant and could be responsible for its haematopoietic and immunomodulatory property.