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AMMELIORATIVE PROPERTIES OF METHANOL LEAF EXTRACT OF MUCUNA PRURIENS ON THE KIDNEY MARKERS OF MALARIA INFECTED MICE
Abstract
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
INTRODUCTION
1.1 Background to 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?
Abstract
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
INTRODUCTION
1.1 Background to 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?
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