Ammeliorative Properties Of Methanol Leaf Extract Of Mucuna Pruriens On The Kidney Markers Of Malaria Infected Mice

Project and Seminar Material for Biochemistry

Ammeliorative Properties Of Methanol Leaf Extract Of Mucuna Pruriens On The Kidney Markers Of Malaria Infected Mice


The research on Mucuna pruriens was to know its ameliorative efficacy on the kidney markers of malaria infected mice. The antimalaria effect was evaluated against Plasmodium berghei stain of 25 mice.The twenty five (25) adult albino mice of both sexes which weighed 20g-36g were grouped into five(1-5) of five mice per group which was used to carry out the experiment. Group 1 which served as negative control received 10ml/kg of 5% tween 80,group 2, which is positive or standard control received 0.95mg/kg of standard drug (ACT), groups 3, 4 and 5 received 100mg/kg, 200mg/kg and 400mg/kg body weight of the extract of Mucuna pruriens respectively. Malaria was induced using Plasmodium berghei in which the malaria infected mice were examined for parasitemia on day 4 and 8 of the treatment.

The creatinine, bilirubin and serum urea levels were determined using Randox and Teco kits .The curative percentage of the animals in groups (1-5) were recorded. In day 4, The curative percentage were 0%, 41.43%, 39.34%, 46.1% and 67.93% respectively and day 8 values are as follows 28.13%, 97.19%, 87.70%, 80% and 95.19%. The values for kidney markers in days 4 for Urea were (44.00 9.79, 36 40 , 35 and 46.67 , creatinine values (0.62 2.62 4.46 3.13 2.53 ) and bilirubin values (21.9 1.41, 21.4 and 22.43 while the values in day 8 for Urea were (40.00 and 45.00 and creatinine values were (24.26 and 23.93 while that of bilirubin were (24.26 and 23.93 for groups (1-5) respectively. The research showed that the extract can cure malaria without affecting the kidney markers and can be added in formulating antimalaria drugs.

Chapter One

1.0 Introduction

1.1 Background of the Study

A medical plant can be described as any plant in which one or more of its organs contains substances that can be used for therapeutic purposes or which are materials for the synthesis of useful drugs (WHO, 2011). Medical plants are very ancient and only true natural medicines have been found useful in several ways. They can be used directly or in other extracted forms for the management of various ailments. They can also be used as agents or starting materials in the synthesis of drugs (Harbone, 2003).

Medical herbs and plant extracts are now generally considered as effective medicines to be respected, appreciated and they play a major role in modern pharmacy (Cunningham, 1993)

The use of herbs is very common in developing countries, particularly in rural settings. However, during the last decade, an increase in the use of plant has been observed in metropolitan areas of development countries (Harnack ,Jeffery and Boutelle, 2001)

Plants are extensively used to treat malaria caused by different species of plasmodium. Malaria is the worldwide most important parasitic disease with an incidence of almost 400-900 millions clinical cases and approximately one to three million death annually (Sachs and Malaney, 2002).

It is endemic in about 100 developing countries. Worse still, malaria parasite have developed defenses against many anti malaria drugs and many insecticide (WHO, 2011). The newly produced effective anti malaria drugs are expensive, especially for most poor Nigerians to afford. Chemotherapy with effective anti malaria drugs remain the main method to control of malaria in the absence of a suitable vaccine treatment (WHO, 2011). Coupled with the human preference for natural therapies, the knowledge of traditional medicine to combat malaria and other fevers was widely diffused among the needed for these traditional remedies were accessible free at the background or at very cheap rates and can be self prepared at home. Malaria is a disease caused by parasitic microbes (plasmodium species) spread by the female anopheles mosquitoes (Fisher and Bialek, 2002).

Plasmodium falciparum resistance to commonly used anti-malaria drugs is rising rapidly in Nigeria and other parts of Africa (Collins and Barnwell, 2009). This has resulted in insurgence in transmission and an increase in adverse outcomes due to therapy failure. In fact, this loss of effectiveness of chemotherapy constitutes the greatest threat to the control of malaria. He world Health Organization has estimated that funding for malaria control alone, including only existing method for vector control, will need to increase to 3.1billion annually by 2015 (WHO, 2004). Effect of plasmodium falciparum varies from a symptomatic to multi organ manifestation, which could lead to death of victim (Zaki, et al., 2013). Malaria has been implicated as one of the factors responsible for renal and hepatic dysfunction in malaria endemic area countries (Mishra et al., 2013, Ogbadoyi et al., 2007, Sharma et al., 2004). The malaria parasites usually affect the kidney, liver and brain (Dzeing-Ella et al., 2005).

The level of severity of malaria infection can be determined by both renal and hepatic malfunction. The clinical manifestation of renal involvement is associated with infection by plasmodium falciparum and plasmodium malariae (Naqvi et al., 2003), and may be responsible for an immune complex medicated glomerular disease leading to nephritic syndrome. Other implications range from urinary sediment abnormalities, mild protienuria and electrolyte changes acute renal failure with metabolic acidosis (Padhi and Mishra, 2012).

Malaria has been reported to be one of the factor responsible for acute renal failure among children in malaria endemic areas (Mockenhaupt, Beeson and Marsh, 2004), and this adverse effect of malaria parasite on the kidney could lead to an increase in blood urea, hypernareamia, hyper-Kalaemia, low urine specific gravity, metabolic acidosis and low ratio of urinary to blood urea (Padhi and Mishra, 2012)

The sudden increase in the urea level and imbalance in the electrolytes level such as sodium, potassium, bicarbonate and chloride in malaria infected people could serve as indicators for kidney dysfunction (Uzuegbu, 2011, Ebele et al., 2010, Jasani, Hellgren and Rombo, 2012).

Mucuna pruriens is a tropical legume known as velvet beans, devil beans, Mucuna, Nescafe and buffalo bean. Other common names includes Yerepe (Yoruba) and Agbala (Igbo) (Uzuegbu, 2011)

The leaves of Mucuna pruriens are also used in the management of ulcer, cephalgia and general debility. The seeds have been known to contain large amount of protein and mineral and with high calorific value, but also with high level of antinutritive properties such as phenolics, tannins,L-dopa, trypsin inhibitors, and phytohaemaglutinins. The seed also contain glutathione lecithin, gallic acid, nicotine, prurenidine and 4-tetra isoquinoline alkaloids (Mishra and Wagner, 2004) and as such are used as astringent, laxative, antiheminthic, aphrodisiac in the cotyledon of the seed, was found to increase the brain mitochondria complex activity and thus has been attributed to the effects as such has been useful in the management of parkinson’s disease (Jacobsen, 1993).

1.2 Statement of the Problem

Even though pharmacological industries have produced a number of new antimalaria drugs in the last three decades, resistance to these drugs by microorganisms have increased. Current work was undertaken to investigate the ameliorative efficacy of the extract of the leaves of Mucuna pruriens on kidney markers of malaria infected mice.

1.3 Aims / Objectives of the Study

This includes to get extract from the leaves of Mucuna pruriens using methanol and subsequently to test the ameliorative efficacy of the extract on the kidney markers of malaria infected mice using different doses of the extract.

1.4 Specific Objective

  1. To get extract from the leaves of Mucuna pruriens at Ekwulobia Aguata Local Government Area of Anambra State.
  2. To evaluate the antimicrobial activity of Mucuna pruriens methanol leaf extract on malaria infected mice.
  3. To investigate the effect of the extract on kidney markers of mice.

1.5 Significance of the Study

This research was done to assess therapeutic antimalaria agents from Mucuna pruriens. It was envisaged to lay down the groundwork for enhancing traditional knowledge and practices through modern approaches of drug development. The study is also aimed at contributing to scientific knowledge locally, nationally and internationally in the area of malaria disease.

The result of this study will determine the antimalaria activity of Mucuna pruriens to plasmodium species involved in malaria. And if the results are favourable then consumption of Mucuna pruriens will be advocated, which is safer than consumption of medical drugs which often have side effect to the individuals taking them, as a preventive therapy (since it raises the immune system of the individuals involved).

1.6 Scope of the Study

This includes obtaining the leaf samples from Ekwulobia Aguata Local Government Area of Anambra State, preparing the samples for extraction and extraction using methanol ,subsequently the ameliorative efficacy of the extract will be tested on kidney markers of malaria infected mice using Plasmodium breghei parasite and 25 albino mice.

1.7 Research Questions

  1. How can extract be obtained from the leaf samples of Mucuna pruriens obtained from Ekwulobia Aguata Local Government Area of Anambra State?
  2. How can the antimalaria activity of Mucuna pruriens methanol leaf extract on malaria infected mice be evaluated.
  3. How can the effect of the extract on kidney markers of mice be investigated?

Chapter Five

5.0 Discussion, Conclusion and Recommendation

5.1 Discussion

From table 2 the results showed that percentage parasitemia of the mice treated with methanol leaf extract of Mucuna pruriens increased significantly in group 2 (positive control) when compared to group 1 (negative control and other groups. Group 3 (100mg/kg body weight of the extract), group 4 (200mg/kg body weight showed no significant difference (p >0.05 in percentage parasitemia compared to group 1 (negative control)

Parasitamia and renal markers were then daily monitored. It was found that BUN (blood Urea Nitrogen) and creatinine levels were markedly increased in the response to the presence of the parasite, which reached significant values firstly on day 4 after infection (tables 3, 5 and 7).

The infected mice were given 100, 200 and 400 mg/kg of the Mucuna pruriens methanol leaf extract for 4 consecutive days subsequently with measurement of renal markers.

As shown in tables 3, 5 and 7, during malaria infection, renal damage was developed as indicated by increasing of BUN and creatinine levels non significantly (P >0.05). interestly, Mucuna pruriens methanol leaf extract exerted a dose dependent protection of renal damage induced by infection as indicated by reduction of BUN and creatinine levels in the extract treated groups. Significant protection were observed at doses of 100, 200 and 400mg/kg of Mucuna pruriens methanol leaf extract, compared with untreated groups, and no significances were found when compared to normal groups. Moreover, prolonged survival time was also observed in infected mice treated with 100, 200 and 400 mg/kg of the extract. Additionally, no toxic effects were found in normal mice treated with 400mg/kg of this extract.

In the current study, we provide evidence that malaria associated renal injury could be protected against by using the Mucuna pruriens methanol leaf extract in malaria infected mouse model.

Impairment of renal function during malaria infection has been reported by clinical reports and it is an important life threatening complication of malaria infection that goes beyond the classical clinical symptoms of malaria. Moreover, the adversities to access of medical services or delay in diagnosis in their place of origin is implicated in the severity of disease. The onset of renal injury in malaria infected mice came out from day 4 after infection and the incidence of renal injury was confirmed through manifestation including marked increases of plasma BUN and creatinine levels. According to Collins and Barnwell (2009) pathophysiology of malaria infection was usually associated with an activation of immune system and comprehends a complex network with production of oxidative stress and inflammation. It has been described that the erythrocyte destruction during blood stage of infection accumulated high levels of toxic free heme in circulation that had the ability to induce oxidation stress from production of hydroxyl radicals via the Fenton/Haber Weiss reaction and resulting renal injury. In a study by Kao et al (2012), it was shown that apoptosis of renal tissues during malaria infection has also been suggested in renal injury. Additionally, pathogenesis of malaria associated renal injury is most likely to be due to immune complex mediated glomerulonephritis caused by immune complex deposition and endothelial damage, which may lead to fatal forms of malaria nephropathies.

Tables 3, 5 and 7 shows the values of some biochemical parameters indicating the kidney function y=test. The normal control group had a value of 3.15 mg/dl. Those treated with the plant extract (3, 4 and 5) had increased creatinine values of 5.55, 4.10 and 3.70 for groups 3,4 and 5 respectively, which was significantly different from the normal control. Groups treated with standard ACT had reduced creatinine values of 2.5mg/dl, although not significantly different (p>0.05) when compared with control group.

The urea value was significantly (p<0.05) reduced in group having a value of 25mg/dl when compared to other group. The group treated with the plant extract had increased uric acid concentration, other treated group was also reduced. All the infected groups had elevated urea concentration. About seven fold increase was observed in treated group when compared to the normal control group. All the treated groups had increased mean urea values, ranging from 35mg/dl to 40.00mg/dl and these were significantly different (p<0.05) compared to the normal control group (NC).

5.2 Conclusion

The present study here has demonstrated that Mucuna pruriens methanol leaf extract non significantly reduced plasma BUN and creatinine levels as indicators of renal injury induced by malaria leaf extract on renal injury may be due to its unique composition, where the Mucuna pruriens methanol leaf extract is rich in flavonoides (rutin and quercetin, flavonoglycosides, polyphenols, and tannins).

5.3 Recommendation

This study have shown that Mucuna pruriens methanol leaf extract is most effective at the dose levels of 100, 200 and 400mg/kg. This plant can be recommended for use since it possessed a high protective effect against malaria and can be obtained at relatively no cost from nature.

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