CHAPTER ONE
1.0. Introduction
Traditional medicine is the total combination of knowledge whether
explicable or inexplicable use in diagnosing, preventing or eliminating a
physical, mental or social disease which may rely exclusively on past
experience handed down from generation to generation (Boerio-Goates,
2001). A medicinal plant is any plant used for the extraction of pure
substances either for direct medicinal use or for hemi-synthesis of
medicinal compounds which can be used for the therapeutic purpose or as a
precursor for the synthesis of useful drugs (Ponomarev and Migarskaya,
2000),).
The plant Phyllanthus amarus is one of the most important
medicinal plants used in traditional medicine for treatment of diabetes
and excessive body weight reduction.
Its hypoglycemic properties have been reported (Bunn and Higgins, 2011).
Ferritin is a class of iron storage protein distributed in
virtually all living kingdoms. Proteins of this class commonly form
spherical protein nanocages, each of which is usually composed of 24
similar or identical subunits (Oyaizu, 2006). This cage-like 24-mer has a
large inner cavity and shows highly symmetrical architecture, i.e., the
24 subunits are related by four-, three-, and twofold symmetries (432
symmetry). A subunit of ferritin forms a four-helix bundle composed of
helices A to D and a short fifth helix E, which is also a common
structural feature of all ferritins. Many intersubunit interactions have
been observed in the crystal structures of ferritin from various
organisms, giving the ferritin superfamily its super thermal stability
(Krikler and Heathcote, 2002).The ferritin superfamily can be divided
into a vertebrate type, a plant type, a bacterial type, and so on.
Bacterial ferritin can be further divided into heme-containing
bacterioferritin (Bfr), nonheme ferritin (Ftn), and Dps (DNA binding
protein from starved cells), the latter of which exceptionally forms a
dodecameric protein shell with two- and threefold symmetry axes
(mini-ferritin). Most of the ferritins possess a di-iron oxidoreductase
site (ferroxidase site) responsible for iron oxidation in the center of
the four-helix bundle of the monomeric subunit, except for mammalian L
chain ferritin (Rucker et al., 2009).
Glucose is a sugar with the molecular formulaC6H12O6. The name "glucose" (/??lu?ko?s/) comes from the Greek word ???????, meaning "sweet wine, must". Several active compounds have been identified in P. amarus were
found to be potent inhibitors of rat liver cyclic AMP–dependant protein
kinases (Fairclough and Houston, 2004),phyllanthin and hypo-phyllanthin
present are reported to be hepatoprotective agents and protect
hepatocytes against carbon tetrachloride and galactosamine induced liver
toxicity in rats (Gailliot and Baumeister, 2007).
Phyllanthus amarus is a plant of the family Euphorbiaceae
and has about approximately 800 species which are found in tropical and
subtropical countries of the world (Coresh et al., 2007). The name ‘Phyllanthus’ means “leaf and flower” and named so because of its appearance where flower, fruit and leaf appears fused. Phyllanthus amarus is
a branching annual glabrous herb which is 30-60 cm high and have
slender, leaf-bearing branchlets, distichous leaves which are subsessile
elliptic-oblong, obtuse, rounded base (Honeycutt et al., 2013).
Flowers are yellowish, whitish or greenish, auxillary, males flowers in
groups of 1-3 whereas females are solitary. Fruits are
depressed-globose like smooth capsules present underneath the branches
and seeds are trigonous, pale brown with longitudinal parallel ribs on
the back (Williams, 2010).
Phyllanthus amarus is a small erect, annual monoeciousglabous herb that grows to 30-40 cm in height. It belongs to the family Euphorbiaceaewith leaves that alternate distichous and crowded along lateral branchlets (Foo and Wong 2012).
Phyllanthus amarus is a plant with reported medicinal
properties and broad spectrum of pharmacological activities including
antiviral, antimicrobial, anti-plasmodial, anti-inflammatory,
anticancer, antidiabetics, antioxidant and diuretics properties among
others. A number of active constituents of the plant are related to
biologically active lignans, glycosides, flavonoids, ellagitannins and
phenylpropanoids found in the leaf, stem and root of the plant along
with common lipids, sterols and flavonols.[Phyllanthus amarus is a tropical shrub indigenous to the rainforest of Amazon and other tropical areas of the world (Foo, 2005).
Studies on extract of Phyllanthus amarus have shown anti hepatitis B activity, hepatoprotective (Georgieva et al., 2002), anticancerous (Uriezeet al., 2010),
antimicrobial (Samraj (2001) and kidney stones dissolution properties
(Stipanuk, 2000). Therefore, the present investigation was undertaken to
study the ferritin and glucose levels in the serum of mice treated with
ethanoic leaf of exract of P. amarus induced diabetic mice.
1.1. Statement of the Problem
Iron is important for lots of processes in the body; too much iron
over a long period of time can be dangerous and cause a lot of problems
with the function of body organs. Low levels of ferritin are seen in
iron deficiency. Serum insulin levels have been researched in the
general population as potential predictive biomarkers for all-cause and
cause-specific mortality, but initial research has found mixed results.
Subsequent studies have suggested that the more stable insulin precursor
molecule, proinsulin, is a better predictive marker of all-cause and
cause-specific mortality, in particular, cardiovascular mortality
(Blacklocket al., 2000). However, there have been few long-term
prospective studies to evaluate this finding. Proinsulin and mortality
from cancer has not been studied.
Raised serum ferritin can be caused by five main mechanisms: damage
to ferritin-containing tissues, for example, the bone marrow and liver;
inflammation or infection, because ferritin is an acute-phase protein;
genetic iron-loading conditions; secondary iron-loading conditions,
which are mainly due to blood transfusions but are also seen in African
haemosiderosis due to the ingestion of large quantities of iron; and
chronic anaemias caused by ineffective haematopoesis, for example,
thalassaemias elimination (Hasratet al., 2004).
Low serum ferritin is a common problem in people with gluten intolerance.
In celiac disease, antibodies first target the proximal small intestine
(the earlier part of your small intestine), which is the part of your
small intestine responsible for absorbing iron. Some forms of non-celiac
gluten sensitivity (NCGS) also trigger antibodies that do damage to
this part of your intestine, while other forms of NCGS trigger systemic
inflammation, which may impede iron absorption. Low ferritin is most
common and most severe in celiac disease patients (Koffi, 2007).
Diabetes mellitus (DM) is caused by inherited and/ or acquired
deficiency in production of insulin by the pancreas, or by the
ineffectiveness of the insulin produced . This insulin deficiency
results in increased concentration of glucose in the blood. Increase in
blood glucose damages many of the body’s systems, in particular, the
blood vessels and nerves. The hyperglycemia caused due to decreased
insulin production is called Type-1 diabetes and hyperglycemia due
insufficient insulin utilization is called Type-2 diabetes. Out of these
two types, Type -2 diabetes is a major problem of today and it account
for nearly 95% of total diabetic population of about 246 million. Since
ancient times, plants have been an exemplary source of medicine (Konan et al., 2006).
Phyllanthinis reported to be hepatoprotective agents and protect
hepatocytes against carbon tetrachloride and galactosamine induced liver
toxicity in rats (Syamasundaret al., 1985). In Nigeria, the
plant is extensively used in traditional medicine to eliminate waste
from the body, restore the activity of the liver and build up blood and
innate defense system.
Phyllanthus amarus is a small herb common to central and
southern India. It can grow to 30-60 cm in height and bloom with yellow
flowers. All parts of the plant are used in ayurvedhic medicines because
of their medicinal properties. Leaves of this plant are reported to
contain lignans, alkaloids, flavonoids, galloatnoids and glycosides
It is noted that evaluation of this herb and herbal products in
general faces major problems. First, is the use of mixed extracts
(concoctions) and variations in methods of harvesting, preparing, and
extracting the herb, which can result in dramatically different levels
of certain alkaloids.
Despite the wide spread use of P. amarus much has not been reported about the biochemical effect in Nigeria. A recent work in Nigeria shows that the aqueous extract of P. amarus has hepatic cell function enhancement (Chen et al.,
1996). This work was therefore designed to evaluate ferritin and
glucose levels in the serum of mice treated with ethanoic treated with
extract of P. amarus.
1.2. Objective of the Study
The purpose of this study is to determine the ferritin and glucose
levels in serum of mice treated with ethanolic leaf extract of Phyllantusamarus which may indicate an advantage or disadvantage
1.3. Scope of the Study
The study seeks to evaluate ferritin and glucose levels in the serum mice treated with ethanoic leaf extract of phyllantusamarus using mice model. A group of male mice induced with ferritin and glucose would be treated with leaf extract of phyllantusamarus and
also would be treated with the juice extract of the plant on daily
basis and another group who are otherwise healthy would serve as a
positive control while an induced but not treated group would serve as a
negative control for the experiment. The research will be carried out
in the Animal Facility of the Faculty of Basic Medical Sciences of Delta
State University Abraka.
1.4 Significance of the Study
The result of this study may be used to help determine the cause of
hypoglycaemia (low glucose). The findings of this study will be of
traditional usefulness in several health problems such as diarrhoea,
dysentery, dropsy, jaundice, intermittent fevers, urinogenital
disorders, scabies and wounds. This study will enlighten riverside
communities of the potential danger they face and how to control it
using the Phyllanthus amarus.
1.5. Limitation of the Study
This study is limited to leaf extract of phyllantusamarus for the treatment of ferritin and glucose levels in serum of mice which may indicate an advantage or disadvantage.