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Science Focus Vol. 18 (1) 2013 pp 57 - 62
Faculty of Pure and Applied Sciences, LAUTECH, Printed in Nigeria
ANTIPLASMODIA EFFICACY OF METHANOLIC EXTRACT OF LEAF
OF MORINDA LUCIDA
Tajudeen O. OSENI and *Kuranga I. AYINLA
Department of Chemistry, Institute of Basic and Applied Sciences, Kwara State Polytechnic, P.M. B. 1375, Ilorin- Nigeria. Abstract
Methanolic leaf extract of Morinda lucida was assessed for antiplasmodial activity against Plasmodiumberghei -infected mice. The phytochemical screening of the methanolic leaf extract showed thepresence of Anthraquines, Alkaloids, Saponins, Tannins and Cardiac glycosides. The results indicatedsignificant (P<0.05) inhibition of parasite at dose levels of 100,200, 400 mg/kg body weight (b.w) ofthe extract and 5mg/kg Chloroquine as seen in the percentage parasite inhibition of 65.36, 75.10 , 82.30and 88.59% respectively. Minimal decrease in the whole body weight and percentage packed cellvolume (% PCV) of the mice were also observed. It can be concluded that methanolic leaf extract ofMorinda lucida is potentially useful for the development of antimalaria drug.
Keywords: Antiplasmodia , Morinda lucida, Methanolic extract, Plasmodium berghei.
Introduction
Malaria is a serious parasitic disease from tropical regions caused by a species of Plasmodium and
transmitted by Anopheles mosquitoes. It is prevalent in approximately 100 countries in Africa,
Southeast Asia and South America, where approximately 2.4 million people are at risk (Kager 2002).
According to the World Malaria Report (WHO, 2009a), there are approximately 250 million malaria
cases and approximately one million people die from malaria each year.
The emergence and rapid spread of multidrug-resistant strains of Plasmodium, particularly Plasmodium falciparum, represent a major problem for prophylaxis and treatment, which becomesmore difficult and limits the choice of drugs used. This has been identified as the current primary causeof control failure.
The well-known use of chloroquine (CQ) and antifolates [sulfadoxine-pyrimethamine (S/P)] formalaria treatment are no longer effective in most endemic areas. Combination therapy has emerged asthe best practical solution in overcoming the resistance of select strains. Therapeutics based oncombinations with artemisinin and derivatives (ACTs), recommended by the World HealthOrganization (WHO), presently represent the most effective treatment of P. falciparum malaria infec-tion (WHO, 2008). Clinical resistance to these combinations has been recently reported in Cambodia(Noedl et al., 2008), suggesting that P. falciparum parasites have already developed the ability to growin the presence of these anti-malarials, which strongly suggests the need for further research into newantimalarials.
*Corresponding author email : Ibkuranga@gmail.com. Tel: +2347065360971
Science Focus: An International Journal of Biological and Physical Sciences
Antiplasmodia efficacy of methanolic extract … There is broad consensus on the urgent need for new, affordable and efficient compounds that couldserve as primary molecules for antimalarial treatment. New highly-effective antimalarial drugcandidates, based on new mechanisms of action or with new structures, are urgently needed toovercome the problem of rapid emergence of drug resistance and achieve long-term clinical efficacy.
Due to the crucial role that plant-derived compounds have played in drug discovery and developmentfor the treatment of several diseases, the isolation of new bioactive compounds from medicinal plantsbased on traditional use or ethnomedical data appears to be a very promising approach ( Turschner andEfferth, 2009).
Morinda lucida ( Rubiaceae) is a medium-sized tree about 15 m tall (Yinusa et al., 2005), it is callednames in different countries. It is known as Sangogo or Bondoukou alongua in Cote d’ Ivoire; Twi, konkroma or Ewe amake in Ghana; Ewe amake or atak ake in Togo and Oruwo in South Western Nigeria(Adeneye and Agbaje, 2008). Different parts of the plants are used in different ways in differentcountries. Cold decoction of the plant leaves is used for the treatment of fever in Cameroon; the bitterwater decoction of the plant bark, root and leaf are used as bitter tonic and as astringent for dysentery,abdominal colic and intestinal worm infestation (Adeneye and Agbaje, 2008). Oliver-Bever (1986)reported the use of weak decoction of the stem bark in the treatment of jaundice. Koumaglo et al(1992) and Obih et al (1985) documented in vitro antimalarial activity of Morinda lucida leaf extractagainst Plasmodium falciparum and antimalarial activity of Morinda lucida against Plasmodiumberghei in mice. Methanolic extract of Morinda lucida leaf extract have been reported to possesstrypanocidal activity (Asuzu and Chineme, 1990).
The antimalaria activities of the leaves of Morinda lucida have been documented (Obih et al., 1985),the present study is designed to evaluate antiplasmodial activities of methanolic leaf extract of Morindalucida in plasmodium berghei- infected mice Material and methods
Sample collection, Preparation and Extraction The plant was collected from Oyun in Ilorin East Local Government of Kwara State. The taxonomicidentity of the plants was verified at the Department of Plant Biology, University of Ilorin, KwaraState.
The plant material was then cleaned, reduced to small fragments, air dried under shade at roomtemperature and coarsely powdered by using a wooden mortar and pestle, the powdered plant materialswere packed in brown envelope and kept until extraction.
70g of the powdered plant material was weighed by sensitive digital weighing balance and extracted in400ml 70% methanol using reflux extractor for 2 hours. After 2 hours, the extract was separated fromthe marc using muslin cloth. The solvent was removed by evaporation under reduced pressure by rotaryevaporator in distillation flask at 45rpm and temperature 65oc to obtain the crude extracts of the plant.
The extract was further concentrated to dryness in a water bath (Emmanuel et al., 2011). The methanolcrude extracts were stored in a refrigerator in air tight plastic containers until used for the study.
Phytochemical screeningThe methanolic leaf extract was screened phytochemically for the presence of its constituents utilizingstandard methods of analyses (Harborne, 1984; and Sofowora, 1993).
After third day (72 hours) of inoculation of the parasite, the mice in the three treatment groups wereadministered with the extracts in doses of 100, 200, 40Omg/kg body weight orally once daily for fourconsecutive days (D1, D2, D3, D4) by dissolving the extract in 1Oml distilled water for each mouse.
Two control groups were used in each experiment, the negative control and positive control groupswhich were administered daily with distilled water and chloroquine, a standard antimalarial drug, at5mg/kg body weight respectively for four consecutive days.
Thin films were made from tail blood from D1 through D4, fixed with methanol and stained withGiemsa for 30 min before being washed, dried and observed under a microscope (Mgbemena, 2010).
Determination of Body Weight Change. The body weight of each mouse in all groups was measured before infection and on D4 by a sensitivedigital weighing balance (Brainweigh 300 balance).
The packed cell volume (PVC) of each mouse was measured before infection and on D4 after infection.
Blood was collected from tail of each mouse in heparinized microhaematocrit capillary tubes up to 3/4thof their length. The tubes were sealed by crystal seal and placed in a microhematocrit centrifuge(Hettich haematokrit) with the sealed ends out wards. The blood was centrifuged at 12,000rpm for5minutes.
Data obtained in this study were analyzed using the statistical software SPSS v16.0 (SPSS Inc,Chicago, Illinois, USA). Numerical data were presented as mean ± standard deviation. The significanceof the mean difference between two independent groups was determined using Student’s t-test, andone-way analysis of variance (ANOVA). A p - value < 0.05 was considered significant.
Results and discussions
The result of Phytochemical Screening of methanolic leaf extracts of Morinda lucida showed thepresence of Alkaloids, Anthraquines, Saponins, Tannins and Cardiac glycosides (table 1).
Table 1: Phytochemical Constituents of Methanolic Leaf Extract of Morinda lucida
Bioactive agents
+++ = High
++ = Moderate
The methanolic leaf extract of morinda lucida contains different classes of secondary metabolites thathave antiplasmodial activity. Alkaloids (Saxena et al., 2003), which have been implicated inantiplasmodial activity of other plants were also detected in morinda lucida extract. The presence ofAlkaloid and other bioactive constituents is likely to have contributed to the observed antiplasmodialactivity of morinda lucida leaf extract.
All the groups infected with plasmodium berghei and treated with the methanolic leaf extract ofmorinda lucida and chloroquine, showed a decrease in parasite count, with chloroquine showing lowestparasite count (4.67± 0.58). However, the negative control groups showed increase in parasite count(40.93± 3.67) (figure 1).
Antiplasmodia efficacy of methanolic extract … Fig.1: Effect of methanolic leaf extract of morinda lucida on parasitaemia of P. berghei infected mice
The in vivo antiplasmodial studies carried out on the methanolic leaf extract have confirmed the workof others on the antiplasmodial activity of extract of the plant parts (leaf, stem bark and root) (Asuzuand chineme, 1990; koumaglo et al., 1992). It has been reported that the stem bark infusion is used asan antimalarial ( Burkill, 1997).
The effect of the crude extract of morinda lucida on percentage PVC on days 0 and 4 is indicated infigures 2. It can be deduced from the chart that there was reduction in PCV of mice in test groupswithin and after the experimental period, except for the groups treated with 200mg/kg where there wasan increase in the PCV level throughout the period.
Fig. 2: Effect of methanolic leaf extracts of morinda lucida on % PCV.
According to Taylor and Hurd, (2001), Plasmodium berghei infected mice suffer from anaemiabecause of RBC destruction, either by parasite multiplication or by spleen reticuloendotelial and actionas the presence of many abnormal RBC stimulates the spleen to produce many phagocytes (Chinchillaet al., 1998). In this study, the extracts of morinda lucida presented significant PVC reduction in adependent manner.
Figure 3 showed a significant (p<0.05) loss of body weight between day 0 and 4 in both positive andnegative control groups and treated groups. Treatment with crude extracts did not prevent body weightloss due to parasitemia.
Fig. 3: Effect the methanolic leaf extract of morinda lucida on the weight of the P. berghei
infected mice.
The methanolic leaf extract of morinda lucida in this study caused significant decrease in body weight.
The loss of body weight in the extract treated mice is possibly due to depressing effect of the crudeextract on feed intake/appetite. In a similar study by Chinchilla et al, (1998), mice treated with crudeextracts of some plants showed a lower body weight pattern as compared with the non-treated oneswhich is in agreement with our results.
Conclusion
The current study suggests that extract from the leaf of Morinda lucida possesses significantantiplasmodial activity as seen in its ability to treat Plasmodium berghei infection in mice. Morindalucida could represent potential source of lead molecule for the development of a new drug fortreatment of malaria. It can be concluded from this study that extract from Morinda lucida leaf ispotentially useful for the treatment of malaria.
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