EVALUATION OF THE ANTIDIABETIC EFFECTS OF WATER AND METHANOLIC EXTRACTS OF AVOCADO (PERSEA AMERICANA) SEED ON ALLOXAN INDUCED DIABETIC RATS | Blazingprojects Postgraduate Thesis
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EVALUATION OF THE ANTIDIABETIC EFFECTS OF WATER AND METHANOLIC EXTRACTS OF AVOCADO (PERSEA AMERICANA) SEED ON ALLOXAN INDUCED DIABETIC RATS

 

Table Of Contents


  • Title Page Certification Dedication Acknowledgement Abstract Table of Contents List of Tables List of FiguresCHAPTER ONE
  • 1.0Introduction
  • 1.1Background
  • 1.2Global Burden of Diabetes
  • 1.3Statement of Problem
  • 1.4Aims and Objectives of the StudyCHAPTER TWO
  • 2.0Literature Review
  • 2.1Diabetes Mellitus 2.
  • 1.1Types of Diabetes Mellitus 2.
  • 1.2Prevention / Treatment of Diabetes Mellitus
  • 2.2Medicinal Plants
  • 2.3Evaluation of Drug Toxicity 2.
  • 3.1Liver Function 2.3.
  • 1.1Total Bilurubin 2.3.
  • 1.2Conjugated Bilurubin 2.3.
  • 1.3Aspartate Transaminase (AST) 2.3.
  • 1.4Alanine Transaminase (ALT) 2.3.
  • 1.5Alkaline Phosphatase (ALP) 2.
  • 3.2Histology of Liver and KidneyCHAPTER THREE
  • 3.0Materials and Methods
  • 3.1Experimental Design
  • 3.2Chemicals and Reagents
  • 3.3Sample Collection and Preparation
  • 3.4Proximate Analysis 3.
  • 4.1Protein Content 3.
  • 4.2Crude Fat Content 3.
  • 4.3Moisture Content 3.
  • 4.4Ash Content Measure 3.
  • 4.5Carbohydrate Content
  • 3.5Anti-nutritional Components 3.
  • 5.1Tannin Content Determination 3.
  • 5.2Oxalate Determination 3.
  • 5.3Phytic Acid
  • 3.6Animal Experiment 3.
  • 6.1Induction of Diabetes 3.
  • 6.2Administration of Extracts 3.
  • 6.3Blood Sample Collection 3.
  • 6.4Liver Function Tests 3.6.
  • 4.1Total Bilirubin Test 3.6.
  • 4.2Conjugated Bilirubin Test 3.6.
  • 4.3Alanine Aminotransferase (ALT) Test 3.6.
  • 4.3Aspartate Aminotransferase (AST) Test 3.6.
  • 4.5Alkaline Phosphatase (ALP) Test 3.
  • 6.5Histological Study 3.6.
  • 5.1Histology Sample Preparation
  • 3.7Statistical Analysis of DataCHAPTER FOUR
  • 4.0Results
  • 4.1Proximate Analysis
  • 4.2Anti-Nutritional Analysis
  • 4.3Animal Experiment Results 4.
  • 3.1Glucose Analysis Results 4.3.
  • 2.Results of Body Weight Analysis 4.3.
  • 3.Liver Function Test Results 4.
  • 3.4Histology ResultsCHAPTER FIVE
  • 5.0Discussion
  • 5.1Proximate and Anti-nutrients
  • 5.2Effects of Extracts on Body Weight
  • 5.3Effects of Seed Extracts on Blood Glucose
  • 5.4Effects of Seed Extracts on Liver Function Parameters
  • 5.5Histological Effects of Extracts on the Kidney and Liver
  • 5.6Conclusion References Appendix

Thesis Abstract

The high prevalence, complications and cost of conventional drugs in the management diabetes necessitated the search for alternative treatment. As a result, this study seeks to evaluate the composition, anti-diabetic potential toxicity and tissue-protective effects of both the water and methanolic extracts of Persea americana (avocado pear) seed on alloxan-induced diabetic albino rats were investigated. This study was conceived and designed based on information on the local use of the seed in diabetes treatment. Proximate and anti-nutritional constituents of the seed were determined and 100g of the sample was extracted with 1000ml of both water and methanol using the maceration method. The extracts were evaporated to dryness using a rotary evaporator and stored at 4oC until use. The effects of different doses (200mg/kg.b.wt., 300/kg.b.wt.) of both water and methanolic extracts of P. americana seed on alloxan-induced diabetic albino rats were compared with those of a reference drug, insulin. The glucose level and weight of the rats were measured weekly for 21 days. The liver function tests and the histopathologies of the liver, and kidneys, were investigated. Results of the proximate investigation shows that the seed is rich in carbohydrate (49.03± 0.02 g/100g), lipid (17.90±0.14 g/100g), protein (15.55± 0.36 g/100g) moisture (15.10± 0.14 g/100g) and ash (2.26±0.23 g/100g). Anti nutritional components such as total oxalate (14.98±0.03 mg/100g), tannin (6.98±0.04 mg/100g) and phytic acid (3.18±0.16 mg/1 00g). Results also showed that both the water and methanolic extracts exhibited significant anti-diabetic effects on the experimental rats. However, the methanolic extracts showed a better anti diabetic effect than the water extracts. The extracts showed no significant effects on the liver function parameters (bilirubin, conjugated bilirubin, AST, ALP and ALT) compared with the normal control but rather reversed the histopathological damage that occurred in alloxan-induced albino diabetic rats. In conclusion, the present study provides a pharmacological basis for the traditional use of P. americana seeds extracts in the management of Diabetes mellitus. It seems P. americana seed contains substantial amount of nutrients that could warrant its utilization in animal feed or food. However, further studies are required to identify the active ingredient responsible for the anti-diabetic properties of the seed extract.

Thesis Overview

INTRODUCTIONBACKGROUNDDuring the past decade, the traditional systems have gained importance in the field of medicine. The World Health Organization estimates that 4 billion people, 80% of the world population (WHO, 2002), presently use herbal medicine for some aspect of primary health care (Orisataoki and Oguntibeju, 2010). Eighty percent (80%) of African populations use some form of traditional herbal medicine (WHO, 2002.) and the worldwide annual market for these products approaches US$ 60 billion (Willcox and Bodeker, 2004). Herbal medicine is a major component in all indigenous people’s traditional medicine. Medicinal plants have continued to attract attention in global search for effective methods of using plants parts (e.g. seeds, leaves, stems, roots, barks etc) for the treatment of many diseases affecting humans (Sofowora, 2008). This is as a result of the continuous need for less expensive means of disease control.
Medicinal plants are plants which can be used for therapeutic purposes or which are precursors for the synthesis of useful drugs (Sofowora, 2008). Many important drugs used in healthcare today are directly derived from plants due to its bioactive constituents such as; alkaloids, tannins, steroids, etc. Examples include L-Dopa derived from Mucuna sppused for anti-parkinsonism, another is Caffeine, a CNS stimulant derived from Camellia sinensis and Quinine from Cinchona ledgeriana and used for Antimalarial, antipyretic.As a matter of fact, well into the 20th century, much of the pharmacopoeia of scientific medicine was derived from the herbal lore of native people. Many drugs commonly used today are of herbal origin. Undisputedly, the history of herbology is inextricably intertwined with that of modern medicine. Many drugs listed as conventional drugs were originally derived from plants.Salicylic acid, a precursor of aspirin, was originally derived from white willow bark and the meadowsweet plant (Zand, et al., 2003).Plants are very unique as their existence is very essential for the sustenance of the rest of the food chain. Based on the observations made through successive generations, superstition as well as traditional medicinal folklore, man has found and has been using herbs, barks, fruits, leaves, seeds, roots and stems of different plants of various climatic regions for therapeutic purposes (Sofowora, 2008).GLOBAL BURDEN OF DIABETESDiabetes mellitus is a metabolic disease caused by a deficiency in the secretion or action of insulin (Nelson and Cox, 2005). This disorder is characterized by major symptoms as; polyuria (frequent and abundant urine), glycosuria (presence of glucose in urine) and hyperglycemia (glucose rate on an empty stomach higher than 1.2g/l in plasma blood and confirmed in at least two occasion) (N’guessan et al., 2009). Basically, there are two major clinical classes of diabetes. The type 1 diabetes or insulin dependent diabetes mellitus (IDDM) and type 2 diabetes or non insulin dependent diabetes mellitus (NIDDM) also called insulin resistant diabetes. According to the International Diabetes Federation (IDF) 2014 updates, out of the world seven billion population, 387million people, aged 20–79 years worldwide are diabetic, (IDF, 2014) giving a comparative prevalence of 8.3%, while 46.3% cases are undiagnosed. In every 7 seconds, a person dies of diabetes, 4.9 million deaths in 2014. Seventy seven percent (77%) of people with diabetes live in low and middle income countries. Africa has recorded cases of 2,150,274 (5.05%) diabetic patients with over 13million undiagnosed cases. In Nigeria, there are estimated 374,651 diabetic cases, with another 172,339 undiagnosed cases. These figures account for about 4.64% Nigerian adults between ages 20-79 living with diabetes (IDF, 2014). In 2014, about 105,090 Nigerians died as a result of diabetes (IDF, 2014). An average diabetic Nigerian spent about 43527.16 naira (US $178.39) in 2014 due to diabetes treatment (IDF, 2014). With this alarming prevalence rate, diabetes mellitus poses a major challenge globally and accounts for a number of disabilities and deaths globally.Currently, diabetes therapy is based on the use of hypoglycemic drugs (sulfonamides, biguanides, and insulin), on hygiene-diet measures, exercise, and requires a lifelong treatment. With the level of poverty in developing nations like Nigeria, the need for a better and cheaper medication cannot be over emphasized. Traditional medicine has always provided a cheaper and time trusted alternative for the treatment and management of various diseases over time.STATEMENT OF PROBLEMThere is an urgent need to provide alternative and cheaper means for the management and treatment of diabetes.AIMS AND OBJECTIVES OF THE STUDYThis study is aimed at the evaluation of the anti-diabetic effects of water and mETHANOLic extracts of Persea americana (avocado) seed on alloxan induced diabetic rats.The specific objectives of the study are:
  1. To determine nutritional and some anti-nutritional constituents of seed extracts of the plant Persea americana.
  2. To evaluate the anti-diabetic properties of seed extracts of americana in diabetic rats
  3. To evaluate the effect of americana seed extract on liver function enzymes.
  4. To investigate the histopathological effects of americana on both the kidney and liver of diabetic rats.
REFERENCESAbbas, Q., Qureshi, I.Z. (2013). Anti-hyperglycemic and Anti-nephropathic Effects of Woodfordia Fruticosa Linn. in Alloxan-induced Diabetic Rats.ISESCO J.of Sci. and Tech.; 9(15), 33-38.Abdel, M.A., El-Feki, M., Salah, E. (1997). Effect of Nigella sativa, fish oil and gliclazide on alloxan diabetic rats. Biochemical and Histopathological studies. J Egy Ger Soci Zool. 1997;23:237–265.Abdulrahman, F.I., Onyeyiri, P.A., Sanni, S., Ogugbuaja, V.O. (2007). Toxic effect of aqueous root bark extract of Vitex doniana on liver and kidney functions. Int. J. of Bio. Chem.;1: 184-195.Adamu A.S., Oyetunde J.G. (2013).comparison of dietary proximate and mineral values of two varieties of bean; Asian Journal of Natural & Applied Sciences 2( 2), 103-106 issn: 2186-8476, issn: 2186-8468 print.Adegoke, G.O., Akinbile, J.T., Olapade, A.A., Ashaye, O.A. (2012). The effect of processing methods on the nutritional profile of avocado (Persea americana Mill) seeds. IJRRAS 11(2) 186-194.Adewole, S.O., Ojewole, J.A.O. (2007). Insulin-induced immunohistochemical and morphological changes in pancreatic β-cells of streptozotocin-treated diabetic rats. Method Find in Exp Clin Pharmacol.;29(7):447–455.Adeyemi, O.O., Opko, S.O. Ogunti, O.O.(2002). Analgesic and anti –inflammatory effects of the aqueous extracts of Persea americana mill (Lauraceae). Fit.; 73; 375-380.Ahmed, O.M., Moneim, A.A., Yazid, I.A., Mahmoud, A.M. (2010). Antihyperglycemic antihyperlipidemic and antioxidant effects and the probable mechanisms of action of Ruta graveolens infusion and rutin in nicotinamide-streptozotocin-induced diabetic rats. Diabeto. Croatica 39:15-35.

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