3.0 MATERIALS AND METHODS
3.1 STUDY AREA
3.2 STUDY POPULATION
3.5 SAMPLE SIZE DETERMINATION
3.6 METHOD OF SAMPLE COLLECTION
METHOD OF SAMPLE ANALYSIS
3.7.1 ESTROGEN ASSAY
3.7.3 REAGENT PREPARATION
3.7.4 ESTROGEN ASSAY PROCEDURE
3.7.6 PROGESTERONE ASSAY PRINCIPLE
PROGESTERONE ASSAY PROCEDURE
3.8 CALCULATION OF
DETERMINATION OF PLASMA FASTING GLUCOSE
METHOD OF STATISTICAL ANALYSIS
1.2 BACKGROUND OF STUDY
mellitus is a group of metabolic disorders in which a person has high plasma
glucose, either because the body does not produce enough insulin, or because
cells do not respond to the insulin that is produced. The high plasma glucose
produces the classical symptoms of polyuria, polydipsia and polyphagia (Rother,
2007). Type 2 diabetes mellitus, formerly non-insulin dependent diabetes
mellitus or adult onset diabetes, is a metabolic disorder that is characterized
by hyperglycemia in the context of insulin resistance and relative insulin
deficiency (Vinay et al., 2008). It
has been regarded as one of the most common metabolic diseases with the rate of
6.4 % in people aged 20-79 years and one of the leading causes of death all
over the world (Burtis et al., 2008;
Vinay et al., 2008; Rosen, 2012).
Nearly 80% of the type 2 diabetes mellitus patients come from developing
countries (Dhindsa et al., 2009).
Many factors, such as genetics, aging and life style, have been involved in the
development of type 2 diabetic mellitus, diagnosis of type 2 diabetic mellitus
are found to be obese (Ramarao and Kaul, 2009). Over 90% of people with
diabetes mellitus are type 2 diabetics and it is reported to be associated with
certain endocrine disorders. There have been increasing anti-diabetic drugs
such as sulfonylureas, biguanides and dipeptidyl peptidase-4 have been used to
control type 2 diabetes mellitus (Murali and Saravanan, 2012; Neerati et al., 2014). However, most of these
anti-diabetic drugs have limited efficacy and many undesirable side effects
such as drug resistance, weight gain, dropsy and high rates of secondary
failure (Tahrani et al., 2010; Murali
and Saravanan, 2012). Therefore, the development of low toxicity, effective and
economic anti-diabetic drugs is still needed and has far-reaching significance.
Metformin, a class of insulin sensitizers, is commonly used for the treatment
of type 2 diabetes. While lowering the blood glucose level, metformin can cause
reduction of fat mass and inhabitation of tumor cell proliferation (Kargulewicz
et al., 2016; Huo et al., 2017). Diabinese is equally used
as an adjunct to diet and exercise to improve glycemic control in adults with
type 2 diabetes mellitus (Murali and Saravanan, 2012).
hydrochloride, a biguanide, is the most popular oral glucose-lowering medication
in most countries, widely viewed as ‘foundation therapy’ for individuals with
newly diagnosed type 2 diabetes mellitus. This reputation has resulted from its
effective glucose-lowering abilities, low cost, weight neutrality, overall good
safety profile (especially the lack of hypoglycaemia as an adverse effect), and
modest evidence for cardioprotection (Inzucchi et al., 2015). A derivative of guanidine, which was initially,
extracted from the plant Galegaofficinalis
or French lilac, metformin was first synthesised in 1922 and introduced as a
medication in humans in 1957, after the studies of Jean Sterne (Sterne, 2007).
Its popularity increased after eventual approval in the USA in 1994, although
it was used extensively in Europe and other regions of the world prior to that
(Pryor and Cabreiro, 2015). The drug’s efficacy has been demonstrated in
monotherapy as well as in combination with other glucose lowering medications
for type 2 diabetes mellitus. Based on these important characteristics, there
continues to be extensive interest in this compound, even many years after its
incorporation into the diabetes pharmacopeia. Interestingly, and despite this
popularity, there still remains controversy about the drug’s precise mechanism
of action, although most data point to a reduction in hepatic glucose
production being predominately involved although, recent data suggests that
some of the drug’s effect may involve the stimulation of intestinal release of
Incretin hormones (Rena et al.,
Diabinese (chlorpropamide) is an oral
blood-glucose-lowering drug of the sulfonylurea class (Inzucchi et al., 2015). It lowers the blood
glucose acutely by stimulating the release of insulin from the pancreas, an
effect dependent upon functioning beta cells in the pancreatic islets. The
mechanism by which diabinese lowers blood glucose during long-term
administration has not been clearly established (Inzucchi et al., 2015). Extra-pancreatic effects may play a part in the
mechanism of action of oral sulfonylurea hypoglycemic drugs.
rest, women have greater storage of free Fatty acids than men, but during
exercise and conditions of sustained increased demand, women were shown to
exert higher oxidation of lipids in relation to carbohydrates. Males instead rely
relatively more on glucose and protein metabolism (Mauvais-Jarvis, 2015). During times of food deprivation, females
reduce energy expenditure with consequent loss of fat stores contrary to males.
can decrease food intake directly by effects in the brain. Tight interacting of
leptin, insulin, neuropeptide Y (NPY), and ghrelin seem to play a vital role (Brown et
al, 2010). Moreover, estrogen can exert direct effects on fat tissue
by enhancing proliferation of pre-adipocytes, especially in females (Anderson et
al., 2001), and by up-regulating sc-2A-adrenergic receptors
promoting SAT accumulation, notably in premenopausal women (Pedersen et
al., 2004). Women display less reduction of insulin sensitivity with
increasing body fat and lower resting energy expenditure, which declines more
rapidly with ageing compared with men. However, up to now, there is paucity of
research that establishes the direct effects of anti-diabetic drugs (metformin
and diabinese) on female sex hormone. This study therefore addresses the gap in
knowledge of the effects of combine anti-diabetic drugs diabinese and metformin
on female sex hormone of patients with type 2 diabetes mellitus.
1.3 AIM OF STUDY
The study aims at evaluating the effects
of metformin and diabinese on female
sex hormone of type 2 diabetes mellitus patients attending University of Ilorin
Teaching Hospital (UITH), Ilorin, Kwara State.
The specific objectives of this
To determine the levels
of oestrogen and progesterone in patients with type 2 diabetes mellitus on metformin and diabinese therapy.
To determine the level of oestrogen and progesterone in apparently
non-diabetic age matched volunteer female individual.
To compare the result of one and two above in order to make inference on
the status of oestrogen and progesterone in type 2 diabetes mellitus patients
To assess the association between the level of Oestrogen and progesterone
and the duration of the diabetes.
1.5 RESEARCH HYPOTHESIS
Anti-diabetic drugs does not have
direct effect on female sex hormone of patients with type 2 diabetes mellitus.
An anti-diabetic drug have direct effect on female sex hormone of patients
with type 2 diabetes mellitus.
OF THE STUDY
this study will be recruited from University of Ilorin Teaching Hospital
(UITH), Ilorin in order to analyse the estrogen and progesterone in serum of
patients with type 2 diabetes mellitus
on metformin and diabinese and compare values with control subjects.
1.7 RESEARCH DESIGN
is a Randomized cross sectional case-control study.
Click “DOWNLOAD NOW” below to get the complete project material
FOR QUICK HELP CHAT WITH US NOW!
+(234) 0814 780 1594