EVALUATION OF THE ANTIHYPERGLYCEAMIC EFFECTS OF POLAR PARTITION FRACTIONS OF HOSLUNDIA OPPOSITA LEAF
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ABSTRACT
Hoslundia opposita leaf is used folklorically in the management of diabetes and a preliminary anti-hyperglycaemic activity has been reported for its methanol extract. Therefore, in this study, the antihyperglyceamic activities of the partition fractions of Hoslundia opposita leaf was investigated using glucose loaded wistar rats with a view to determine the most active fraction. A 1% Tween 80 in normal saline and glibenclamide (5 mg/kg) were used as the negative and positive controls, respectively.
The methanolic extract of the plant was successively partitioned into n-hexane, chloroform, ethylacetate, and evaporated to dryness in vacuo to obtain their corresponding n-hexane, chloroform, ethylacetate and aqueous fractions, which were similarly tested for their anti-hyperglyceamic activities. Activities of the fractions were statistically compared with themselves, and those of the positive and negative controls using ANOVA, followed by Student Newman Kuels’ test to determine the most active fraction.
The methanolic extract of H. opposita (200, 400 mg/kg) gave a similar profile of activity with glibenclamide probably indicating both extra-pancreatic and insulin release effects. The similar results given by 200 and 400 mg/kg confirmed that the constituents are probably acting in synergism as the extract was significantly more active than the partition fractions or the most active constituents are in the non-polar fractions.
The activities of the fractions were significantly less than that of the standard drug used, glibenclamide (5 mg/kg). The comparable (p > 0.05) 20 % anti-hyperglycaemic activity given by both ethylacetate and aqueous fractions at 400 and 200 mg/kg, respectively may indicate that both demonstrated moderate anti-hyperglycaemic activity, depending on the dose. Also, since their greatest activity was at 4 h, both fractions may elicit their antihyperglycaemic activity by stimulating insulin release. Only aqueous fraction gave a possible mild extrapancreatic effect. The antihyperglycaemic constituents in these fractions are likely to be moderately polar to polarin nature. Therefore, the totality of the results established the anti-hyperglycaemic activity of H. opposita and its usage in ethnomedicinal management of diabetes.
CHAPTER ONE
1.0 INTRODUCTION
Medicinal plant is any plant, which in one or more of its organs, contains substances that can be used for therapeutic purposes or which are precursors for the synthesis of useful drugs (Sofowora, 2008). The practice of using plants in treating diseases is called herbal medicine and it dates back to very earliest periods of known human history. There is evidence of the use of herbs in the treatment of diseases and for revitalising body systems in almost all ancient civilisations – The Indian, Egyptian, Chinese and even the Greek and Roman civilisations (Bakhru, 1992).
The universal role of plants in the treatment of diseases is exemplified by their employment in all the major systems of medicine, irrespective of the underlying philosophical premise (Evans, 2008). Examples of different diseases that plants have been used to treat include heart disorders, digestive disorders, constipation, diarrhoea, gonorrhoea, skin diseases, stomach ache, craw craw, asthma, toothache, hypertension (Bakhru, 1992; Evans, 2008). Some drugs of importance for the treatment of various diseases, which have been obtained from plants include atropine, bromelain, caffeine, cocaine, codeine, digoxin, emetine, morphine, noscapine, pilocarpine and quinine (Ertug, 2000).
1.1. DIABETES MELLITUS
Diabetes mellitus (DM) is a chronic, progressive group of disorders characterised by a relative or absolute deficiency of insulin secretion or peripheral insulin resistance resulting in hyperglycaemia and impaired metabolism of fats, carbohydrates and proteins (Olaniyi, 2005). It is a chronic disorder of carbohydrate, lipid, and protein metabolism typified by persistent elevations (> 200 mg/dL) of fasting blood glucose, due to insufficient or complete cessation of insulin synthesis or secretion and/or peripheral resistance to insulin action (Murray and Pizzorno, 1997).
The disease is associated with increased risk of heart disease, stroke, kidney disease, retinopathy, neuropathy, ulceration and gangrene of extremities (Rotshteyn and Zito, 2004). Diabetes mellitus is a chronic metabolic disorder characterised by a high blood glucose concentration (hyperglycaemia) and a fasting plasma glucose (≥ 7.0 mmol/L) or plasma glucose (> 11.1 mmol/L) two hours after a meal, caused by insulin deficiency, often combined with insulin resistance.
Hyperglycaemia occurs because of uncontrolled hepatic glucose output, reduced uptake of glucose by skeletal muscle and reduced glycogen synthesis. When the renal threshold for glucose re-absorption is exceeded, glucose spills over into the urine (glycosuria) and causes an osmotic diuresis (polyuria), which in turn, results in dehydration, thirst and increased drinking (polydipsia). Insulin deficiency causes wasting through increased breakdown and reduced synthesis of proteins (Rang et al., 2007). Diabetes mellitus is one of the most important non communicable diseases in Nigeria and is second only to hypertension in terms of public health significance. The high burden of DM in Nigeria is largely attributable to cardiovascular diseases, which account for 15 % of all DM deaths (Ogbera, 2007; Ogbera et al., 2007).
1.1.1. Classification of Diabetes Mellitus
(a). Type 1 Diabetes
Type 1 diabetes mellitus is caused by the autoimmune destruction of the β-cells of the pancreatic islets (Taylor, 1999). It is a disease resulting from absolute insulin deficiency, usually caused by autoimmune destruction of pancreatic islet cells. The initial clinical presentation may be ketoacidosis with an acute illness, or a more gradual presentation with symptoms of hyperglycaemia. Other autoimmune disorders may also be present such as Addison’s disease, thyroiditis and pernicious anaemia (Federal Bureau Prisons of Clinical Guidelines, 2008).
A small subset of patients with type 1 diabetes has a non-immune mediated disease process with a waxing and waning clinical course and this form of type 1 diabetes is strongly inherited and most commonly affects persons of African and Asian descent (Federal Bureau Prisons of Clinical Guidelines, 2008). Type 1 diabetic patients are usually young (children or adolescents) and not obese when they first develop symptoms (Rang and Dale, 2007). As in all autoimmune diseases, genetic susceptibility and environmental factors play important role in pathogenesis of type 1 DM (Kumar et al., 2005).
(b). Type 2 Diabetes
This form of diabetes was previously referred to as non-insulin-dependent diabetes (NIDDM) or adult-onset diabetes. Type 2 diabetes is characterised by tissue resistance to the action of insulin combined with a relative deficiency in insulin secretion. A given individual may have more resistance or β-cell deficiency and the abnormalities may be mild or severe (Katzung, 2006) It is likely that 10–20 % of individuals in whom type 2 diabetes was initially diagnosed actually have both type 1 and type 2 or a slowly progressing type 1, and ultimately will require insulin replacement (Katzung, 2006).
Although insulin is produced by the β-cells in these patients, it is inadequate to overcome the resistance and the blood glucose rises (Katzung, 2006). In type 2 diabetes, the amount of insulin produced is not enough or cells of the body do not respond to its presence. It is usually associated with normal B-cell morphology and insulin content, if the β-cells have not become exhausted. (Singh et al., 2010).
(c). Gestational diabetes (GDM)
Gestational Diabetes GDM is defined as glucose intolerance of variable degrees with onset or first recognition during the present pregnancy (Jovanovic, 2001). It is diagnosed in approximately 4% of all pregnancies in the USA (Katzung, 2006). When maternal glucose is not normalised, the outcome of pregnancy is not normal. Therefore, identifying the woman at risk for an abnormal outcome of pregnancy is based on maternal blood glucose (BG) levels (Jovanovic, 2001).
During pregnancy, placenta and placental hormones create insulin resistance that is most pronounced in the last trimester (Katzung, 2006). However, women with a history of GDM have an increased risk for future glucose intolerance, as manifested by maternal diabetes or by recurrent GDM in subsequent pregnancies
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