Diabetes is one of the main health problems in our society mostly in obese or overweight patients. It may be due to very high or very low glycemic in the body. There are two types of diabetes: Diabetes Insipidus (Type 1) and Diabetes Melllitus (Type 2). Diabetes Mellitus is the most common type because it affects mostly the adults. Three common symptoms are seen in a diabetic person these are polyuria, polydipsia and polyphagia. Diabetes is a very common disease and you will carry this disease for the rest of your life. It is treated with insulin. Insulin helps in lowering the blood sugar level of a diabetic person.
The rapidly increasing incidence of diabetes mellitus is becoming a serious threat to mankind health in all parts of the world. However, because of low income status or inaccessibility to health care facilities, many Filipinos favor the use of traditional herbal medications because of its cheap cost and easy accessibility. Annona muricata L. (guyabano) is a tropical plant species known for its edible fruit which has some medicinal merits.
A. muricata were shown to possess anti-stress, anti-inflammatory, contraceptive, anti-tumoral, anti-ulceric, wound healing, hepato-protective, anti-icteric and hypoglycemic activities. In addition, clinical studies support the hypoglycemic activity of the ethanolic extracts of A. muricata leaves. Previous research showed that bark of plants soursop has a most excellent efficacy compared with the roots and leaves of the Annona muricata in lowering blood glucose levels in male rats induced streptozozin (Rahmawati, 2014).
The researchers are going to use ICR mice as experimental models as mice are most widely used to establish models in research. The soursop fruit is known to reduce blood sugar level in ICR mice but this study aims to lower the blood sugar level of the diabetic ICR mice by using Annona muricata leaves extract.
1.2 Objectives of the Study
General
To determine the significant change in the blood sugar level of the diabetes-induced ICR mice before and after administration of the Annona muricata supercritical fluid leaf extract.
Specific
To determine the notable change in the blood sugar level of diabetes-induced ICR mice when administered with different levels of concentration of the Annona muricata supercritical fluid leaf extract.
To lower the blood sugar of diabetes-induced ISR mice with the use of Annona muricata supercritical fluid leaf extract.
1.3 Statement of the Problem
In every research study there are problems that needed to be solved by the researchers.
These are the following questions that the researchers wished to answer.
General
Is there a significant change in the blood sugar level of diabetes-induced ICR mice before and after administration of Annona muricata supercritical fluid leaf extract?
Specific
Is there a notable change in the blood sugar of diabetes-induced ICR mice when administered with different levels of concentration of the Annona muricata supercritical fluid leaf extract?
1.4 Hypothesis
Null Hypothesis
There is no significant change in the blood sugar level of diabetes-induced ICR mice before and after administration of Annona muricata supercritical fluid leaf extract.
There is no notable change in the blood sugar level of diabetes-induced ICR mice when administered with different levels of concentration of the Annona muricata supercritical fluid leaf extract.
Alternative Hypothesis
There is a significant change in the blood sugar level of diabetes-induced ICR mice before and after administration of Annona muricata leaf supercritical fluid extract.
There is a notable change in the blood sugar level of diabetes-induced ICR mice when administered with different levels of concentration of the Annona muricata leaf extract.
1.5 Significance of the Study
The primary aim of this study is to determine if there is a significant change in the blood glucose level of ICR mice using Annona muricata supercritical leaf extract. The result of this study will significantly help the following:
To diabetic patients, the result of this study will supplement the knowledge regarding the benefits of Annona muricata leaves as a natural hypoglycemic agent.
To pharmaceutical field, the study may also contribute to obtaining a sustainable and efficient alternative hypoglycemic agent to synthetic hypoglycemic agent.
1.6 Scope and Limitations
Scope
The study will be conducted for the purpose of determining the hypoglycemic activity of the Anonna muricata supercritical fluid leaf extract in diabetic induced ICR mice.
- The method of extraction of Anonna muricata leaves will be through supercritical fluid extraction.
- Alloxan will be used as diabetogenic drugs in ICR mice.
- Fasting blood glucose will be performed using glucometer.
Limitations
The study will only use sixty live mice. No other species of rats shall be used.
The glucose level of the mice will be the only measured blood component.
The study will only use Annona muricata leaves. No other portions of Annona muricata shall be used.
1.7 Locale of the Study
The researchers will be collecting Anonna muricata leaves at Pinagdanglayan, Dolores, Quezon. The Supercritical fluid extraction will be at University of the Philippines Los Banos, Laguna and the medical examination of the ICR mice will be held at Department of Science and Technology Taguig City.
Chapter II
Review of Related Literature
2.1 Glucose
2.1.1. Overview
The oxidation of complex organic compounds is relied by the organisms to obtain energy. Carbohydrates are the major food source and energy supply for the body and are stored primarily as liver and muscle glycogen. Disease states involving carbohydrates are split into two groups: hyperglycemia and hypoglycemia. Carbohydrates are compounds containing carbon, hydrogen and oxygen (Bishop, et al., 2013). Glucose is one of the simplest forms of carbohydrate; classified as a monosaccharide. Glucose is a primary source of energy for humans.
The nervous system, including the brain, totally depends on glucose from the surrounding extracellular fluid (ECF) for energy. Nervous tissue cannot concentrate or store carbohydrates; therefore, it is critical to maintain a steady supply of glucose to the tissue. For this reason, the concentration of glucose in the ECF must be maintained in a narrow range. When the concentration falls below a certain level, the nervous tissue loses the primary energy source and is incapable of maintaining normal function. (Stoker, 2016)
2.1.2 Hormones
The liver, pancreas, and other endocrine glands are all involved in controlling the blood glucose concentrations within a narrow range. During a brief fast, glucose is supplied to the ECF from the liver through glycogenolysis. When the fasting period is longer than 1 day, glucose is synthesized from other sources through gluconeogenesis. Control of blood glucose is under two major hormones: insulin and glucagon both produced by the pancreas. Their actions oppose each other. (Bishop, et al., 2013)
Insulin is the primary hormone responsible for the entry of glucose into the cell. It is synthesized by the b-cells of islets of Langerhans in the pancreas. When these cells detect an increase in body glucose, they release insulin. The release of insulin causes an increased movement of glucose into the cells and increased glucose metabolism. Insulin is normally released when glucose levels are high and is not released when glucose levels are decreased.
It decreases plasma glucose levels by increasing the transport entry of glucose in muscle and adipose tissue by way of nonspecific receptors. It also regulates glucose by increasing glycogenesis, lipogenesis, and glycolysis and inhibiting glycogenolysis. Insulin is the only hormone that decreases glucose levels and can be referred to as a hypoglycemic agent (Bishop, et al., 2013)
Glucagon is the primary hormone responsible for increasing glucose levels. It is synthesized by the a-cells of islets of Langerhans in the pancreas and released during stress and fasting states. When these cells detect a decrease in body glucose, they release glucagon. Glucagon acts by increasing plasma glucose levels by glycogenolysis in the liver and an increase in gluconeogenesis. It can be referred to as a hyperglycemic agent (Bishop, et al., 2013).
2.1.3 Carbohydrate metabolism
Carbohydrate metabolism begins with digestion in the small intestine where monosaccharides are absorbed into the blood stream. Blood sugar concentrations are controlled by three hormones: insulin, glucagon, and epinephrine. If the concentration of glucose in the blood is too high, insulin is secreted by the pancreas. Insulin stimulates the transfer of glucose into the cells, especially in the liver and muscles, although other organs are also able to metabolize glucose. In the liver and muscles, most of the glucose is changed into glycogen by the process of glycogenesis (anabolism).
Glycogen is stored in the liver and muscles until needed at some later time when glucose levels are low. If blood glucose levels are low, then epinephrine and glucagon hormones are secreted to stimulate the conversion of glycogen to glucose. This process is called glycogenolysis (catabolism). If glucose is needed immediately upon entering the cells to supply energy, it begins the metabolic process called glycolysis (catabolism). The end products of glycolysis are pyruvic acid and ATP. Since glycolysis releases relatively little ATP, further reactions continue to convert pyruvic acid to acetyl CoA and then citric acid in the citric acid cycle.
The majority of the ATP is made from oxidations in the citric acid cycle in connection with the electron transport chain. During strenuous muscular activity, pyruvic acid is converted into lactic acid rather than acetyl CoA. During the resting period, the lactic acid is converted back to pyruvic acid. The pyruvic acid in turn is converted back to glucose by the process called gluconeogenesis (anabolism). If the glucose is not needed at that moment, it is converted into glycogen by glycogenesis. You can remember those terms if you think of \"genesis\" as the formation-beginning (Ophardt, 2003).
2.2 Diabetes
According to International Diabetes Federation, in the year 2017, there are 425 million people who have diabetes around the world while there were more than 3,721,900 diabetic patients in the Philippines. Diabetes is degenerative disease characterized by elevated glucose levels in the blood. The classical symptoms of an uncontrolled diabetic condition are polyuria (excessive urination), polyphagia (excessive hunger), and polydipsia (excessive thirst).
There are several complications including microvascular problems such as nephropathy, neuropathy, and retinopathy. From the Greek words, “diabetes” meaning “siphon” and “mellitus” means “sweet”. Way back second century A.D., a Greek physician named Aretus the Cappadocian discovered diabetes from his observation in some people was in a state which their body acts like a siphon, taking water in at one end and discharging it at the other, and the urine has a sweet taste. Diabetes plays an essential role in oxidative stress and abnormal lipid metabolism that results in dyslipidemia. (Stoker, 2016)
2.2.1 Types of Diabetes Mellitus
2.2.1.1 Type I Diabetes
Type I is also called as Insulin-dependent Diabetes Mellitus (IDDM). It constitutes about 10% of all diabetic cases, and common to children and adolescents. It resulted to an inadequate insulin production caused by the autoimmune destruction of insulin-producing beta cells in the islets of the pancreas. When the hyperglycemia develops in the body, mostly the beta cells are destroyed. The antibody markers, islet cell autoantibodies, insulin autoantibodies, glutamic acid decarboxylase autoantibodies, and tyrosine phosphatase IA-2 and IA-2B autoantibodies, for the destruction of beta cells are present before and during onset of diabetes.
There should be at least 2 autoantibodies to be detected for it will have a greater risk of having type 1 diabetes. The levels of C-peptide and endogenous insulin are either low or undetectable. Patients with untreated IDDM for long periods of time may develop diabetic ketoacidosis. The therapy for this type 1 involves injecting insulin and special dietary programs. Too much insulin injections lead to severe hypoglycemia or insulin shoc