Effect of Aqeous Pumpkin (Leaf) Extract 1
Effect of Aqeous Pumpkin (Leaf) Extract 1
Effect of Aqeous Pumpkin (Leaf) Extract 1
1.0 INTRODUCTION
Plants provide an alternative strategy in search for new drugs. There is a rich abundance of
likely that plants will continue to be a valuable source of new molecules which may, after
possible chemical manipulation, provide new and improved drugs ( Shah et al., 2006).
continent specifically for health care, which may also serve as starting materials for drugs
(Sofowora, 1993). Iwu et al. (1999) reported that infectious diseases account for one-half
of all deaths in the tropical countries. As a result, people of all continents have long
applied poultice and imbibed infusions of indigenous plants dating back to prehistory for
hundreds of years before the development of modern scientific medicine and is still in use
According to the World Health Organization (WHO, 1977) a medicinal plant is any plant
which in one or more of its organ contains substances that can be used for the synthesis of
useful drugs. This definition distinguishes those plants whose therapeutics properties and
constituents have been established scientifically and plants that are regarded as medicinal
but which have not yet been subjected to thorough investigation. The term herbal drug
determines the part/parts of a plant used for preparing medicines (for examples: leaves,
1
plant as herbal preparations produced by subjecting plant materials to extraction,
Medicinal plants contain biologically active chemical substances such as saponins, tannins,
1973; Sofowora, 1996) which have curative properties. These complex chemical substances
of different compositions are found as secondary plant metabolites in one or more of these
plants. Tyler (1999) has reported that plants also contain certain other compounds that
oyster nut are often erroneously used for Telfairia occidentalis. Cultivars of Telfairia
vigour, days to flowering and succulence. In Nigeria the two main cultivars are ugu-ala,
characterized by succulent, broad leaves, small black seeds, thick stem and slow growth
and ugu-elu which has a high growth rate, large brownish seeds with high viability and thin
stem with small leaves. The large succulent leaves of ugu-ala make this cultivar a
commercial vegetable in high demand, while the fast emergence and high growth rate of
Telfairia occidentalis commonly called fluted pumpkin occurs in the forest zone of West
and Central Africa, most frequently in Benin, Nigeria and Cameroon. It is a popular
2
vegetable all over Nigeria. It is rare in Uganda and absent in the rest of East Africa. It has
been suggested that it originated in south-east Nigeria and was distributed by the Igbos,
who have cultivated this crop since time immemorial. It is, however, equally possible that
fluted pumpkin was originally wild throughout its current range, but that wild plants have
been harvested to local extinction and are now replaced by cultivated forms. It has different
traditional names; among the Igbos, it is known as Ugu, iroko or aporoko in Yoruba, ubong
in Efik, umee in Urhobo and umeke in Edo ( Akoroda, 1990, Badifu and Ogunsina, 1991).
occidentalis which by far outweighs the little possible toxicity in biological systems that
Synthetic drugs have over time been associated with terrible adverse effects while plants
have been shown to possess protective and therapeutic properties with minimal adverse
1.3 JUSTIFICATION
Telfairia occidentalis is a widely known vegetable in Nigeria and research as shown that it
examining the effect of aqueous Telfairia occidentalis leaf on the liver of rats exposed to
3
1.4 OBJECTIVE OF THE STUDY
This study will enable the researchers to know effect of aqueous pumpkin (leaf) extract on the liver of
4
CHAPTER TWO
Fluted pumpkin occurs in the forest zone of West and Central Africa, most frequently in Benin,
Nigeria and Cameroon. It is a popular vegetable all over Nigeria. It is rare in Uganda, and absent in
the rest of East Africa. It has been suggested that it originated in south-east Nigeria and was
distributed by the Igbos, who have cultivated this crop since time immemorial. It is, however, equally
possible that fluted pumpkin was originally wild throughout its current range, but that wild plants
have been harvested to local extinction and are now replaced by cultivated forms.
Cultivars of Telfairia occidentalis are distinguished by seed colour, thickness ofvine, size of leaf,
growing vigour, days to flowering and succulence. In Nigeria the two main cultivars are ‘ugu-ala’,
characterized by succulent, broad leaves, small black seeds, thick stem and slow growth, and ‘ugu-
elu’ which has a high growth rate, large brownish seeds with high viability, and thin stem with small
leaves. The large succulent leaves of ‘ugu-ala’ make this cultivar a commercial vegetable in high
demand, while the fast emergence and high growth rate of ‘ugu-elu’ is preferred by farmers because
of quick returns. The seed is often polyembryonic, which is useful for multiplication and in breeding.
It is Perennial, dioecious herb climbing by coiled, often branched tendrils to a height of more than 20
m; root system ramifying in the top surface of the soil; stem angular, glabrous, becoming fibrous
when old. Leaves arranged spirally, pedately compound with 3–5 leaflets; stipules absent; petiole
5
(2–)4–11(–15) cm long; leaflets with petiolules 0.5–3.5 cm long, central one largest, up to 15(–19)
cm × 10(–12) cm, lateral ones asymmetrical, usually dentate in the upper two-thirds, sometimes
scabrid underneath, 3-veined from near the base. Male inflorescence an auxiliary raceme up to 3(–
5.5) cm long, on a peduncle up to 25 cm long, with at base of peduncle one long-pedicellate flower
flowering long before the others; female flowers solitary in leaf axils. Flowers 5-merous, cream
free, oblong, fringed; male flowers with 3 stamens, two 4-locular and one 2-locular, with large
reddish connective; female flowers similar to male flowers but with inferior, cylindrical, 3-celled
ovary and 3 large, heart-shaped stigmas. Fruit a drooping, ellipsoid berry 40–95 cm × 20–50 cm,
weighing up to 6 kg, with 10 prominent ribs, pale green and covered with white bloom wax, fruit pulp
yellow, many-seeded. Seeds compressed ovoid, up to 4.5 cm long, black or brown-red. Seedling with
hypogeal germination, developing first a taproot and then numerous, spreading auxiliary roots;
epicotyl 5–12 cm long; cotyledons plan convex, fleshy. Telfairia occidentalis is a tropical vine grown
in West Africa as a leaf vegetable and for its edible seeds. Common names for the plant include fluted
gourd, fluted pumpkin, ugu (in the Igbo language), and ikong-ubong (in the Efik and Ibibio
southern Nigeria. The fluted gourd grows in many nations of West Africa, but is mainly cultivated in
southeastern Nigeria and it is used primarily in soups and herbal medicines. Although the fruit is
inedible (Ajayi, 1990). The seeds produced by the gourd are high in protein and fat, and can,
therefore, contribute to a well-balanced diet. The plant is a drought-tolerant, dioecious perennial that
in Nigeria, including the Efik, Ibibio, and Urhobo. However, it is predominantly used by the Igbo
6
ethnic group, who continue to cultivate the gourd for food sources and traditional medicines. A
recurring subject in the Igbo’s folklore, the fluted gourd is noted to have healing properties and was
used as a blood tonic, to be administered to the weak or ill. It is endemic to southern Nigeria, and was
Telfairia is a small genus of flowering plants in the squash family which are native to Africa. They
include Telfairia occidentalis, the fluted pumpkin, which is an important vegetable in Nigeria and
other African nations. It is also known as the oyster nut, a common name it shares with its relative
Telfairia pedata. These are woody and herbaceous dioecious vines which bear squashlike fruits that
contain large, nutritious oily seeds. The third species is Telfairiabatesii which is very rare and not
cultivated.
These vines grow quickly to lengths of 30 meters or more, using tendrils to scale trees.
Telfairiapedata is grown as an ornamental plant for its attractive foliage. The seeds of
Telfairiaoccidentalis and Telfairia pedata are over 25% protein and 55% oil, making them a good
source of nutrition. They are used as a traditional food source for nursing mothers, the seed flour is
fermented to make baby food, the leaves and shoots, especially of Telfairia occidentalis , are eaten as
vegetables.
One recent concern has been the impact of Telfairamosaic virus, which, along with other problems, is
The effects of chronic garlic intake on various endogenous antioxidant enzymes and lipid
peroxidation on two major organs, the liver (L) and kidneys (K), were investigated. Wistar albino rats
were fed with fresh garlic homogenate daily by gavage in three different doses (250, 500 and 1000
mg/kg/day) for 30 days. After this period, rats were sacrificed and liver and kidneys were harvested
for biochemical estimation. In comparison to saline-treated rats, the 250 mg/kg/day dose significantly
7
(P<0.02) reduced thiobarbituric acid reactive substances (TBARS) (L: 187.48+/-9.23 vs 150.66+/-
11.45; K: 177.38 15.88 vs 120.66+/-9.39 nmol/g wet. weight) and glutathione peroxidase (GPx) (L:
no change in catalase and reduced glutathione (GSH) but superoxide dismutase (SOD) increased
protein). Both 500 and 1000 mg/kg/day doses significantly (P<0.05) reduced endogenous
antioxidants (catalase and SOD) without altering TBARS. A 1000 mg/kg/day dose of garlic caused
marked histopathological and ultrastructural changes in both liver and kidneys. The results suggest
that garlic in low doses has the potential to enhance the endogenous antioxidant status, although at
higher doses a reversal of these effects is observed. The present study also highlights the potential
ability of a high dose of garlic to induce morphological changes in the liver and kidneys, indicating
Kingdom: Plantae
Class: Tracheophytes
Order: Cucurbitales
Family: Cucurbitaceae
Genus: Telfairia
Species: Telfairiaoccidentalis
2.1.3 ECOLOGY
In the wild, fluted pumpkin occurs in forest fringes and secondary forest, possibly often as a relic of
former cultivation. Fluted pumpkin grows fast in the warm humid tropics, producing edible leaves in
8
the rainy season and at the beginning of the dry season, for a period of 6–10 months. In traditional
agriculture, it is a rainfed crop and water deficiency during the dry season reduces its productivity.
Although it is fairly drought tolerant, rainfall appears to be the major factor in its productivity. The
best leaf and fruit yield and highest plant survival rate occur when the plants are irrigated 2–3 times
per week during the dry season. Fluted pumpkin can be grown under a wide range of soil conditions.
It can be managed as a short-term perennial when grown on well-drained soils, slightly shaded and
mulched. On soggy soils and in sunlit spots it can only be grown as an annual.
White leaf spot disease, caused by Phoma sorghina, reduces the leaf lamina. It also affects the seed. It
is controlled by biweekly foliar spraying with Dithane M-45 at a concentration of 500 ppm. Fusarium
moniliforme forms a dry powdery mass of mycelia on the fruits. Erwinia aroideae causes soft rot of
the leaves with yellowish ooze; it also affects the fruits. A prevalent virus disease is Telfairiamosaic
virus (Telfairiamosaic potyvirus) (TeMV), causing mottling of the leaves and low leaf yield; it also
causes chlorosis, stunting and abnormal fruit development. It is transmitted by the aphid Aphis
spiraecola and via the seed. Telfairia occidentalis is remarkably resistant to root-knot nematodes
(Meloidogyne spp.).
Telfairia occidentalis fruits in storage. Fungi may cause up to 95% loss, bacteria only 5% loss in
foliage and stems. The leaf beetle Copa occidentalis feeds on the leaves, flowers and other plant
and Nezara spp. (green shield bug) feed on leaves, stem and fruits. Margaronia indica defoliates the
9
plant, white beetle (Baris spp.) feeds on fruits. Sylepta derogata, Aphis gossypii and Aphis
spiraecola hinder growth by feeding on stem, foliage and flower buds, and transmit viruses. There are
some unidentified predators that feed on the aphids. Thrips of the genus Taeniothrips cause flower
abortion.
Almost all organisms possess antioxidant defense and repair systems that evolved to protect
them against oxidative damage, these systems are insufficient to prevent them entirely.
human body reduce oxidative damage (Yang et al., 2002). In recent years, there has been a
particular interest in the antioxidant and health benefit of phytochemicals in food and
vegetables. This was as a result of their potential effects on human health ( Wei and Shiow,
2001). Many researchers especially in the field of medical sciences have observed free
green leafy vegetable leaves of Telfairia occidentalis and extracts (such as aqueous and
ethanol extracts) from the leaves have been found to suppress or prevent the production
occidentalis has also be found to protects and ameliorates oxidative brain and liver
10
both aqueous and ethanolic extractof Telfairia occidentalis leaves protect the liver cells
against garlic- induced oxidative damage. However, the aqueous extract is more effective
than the ethanolic extracts, which could be attributed to the higher antioxidant activity of
occidentalis leaves have been reported by Eseyin et al. (2005b), Emeka and Obidoa
(2009) and Kayode et al. (2010). The use of the leaves in folk medicine in Nigeria in the
Malaria is a potentially life-threatening disease in the tropics as it affects over 400 million
people yearly and is responsible for the deaths of an estimated 10,000 women of
reproductive age and over 1 million infants and young children each year ( Barbin,
1989; Mishra et al., 2003). Drug resistance, increases in the production and circulation of
fake drugs and high cost of newer and effective drugs have been a major factor affecting
the poor populace, thus making their choice of herbal remedies inevitable and economical.
A study has shown that the ethanol root extract of Telfairia occidentalis possess
comparable to that of chloroquine (Okokon et al., 2007). The ethanolic and aqueous
extracts of Telfairia occidentalis have been reported to show inhibitory effect on growth on
did not inhibit the growth of the fungi tested, which are Aspergillus favus, Aspergillus
11
activity of the leaves was also reported by Odoemena and Onyeneke
In Nigeria, the herbal preparation of the plant has been employed in the treatment of
anaemia, chronic fatigue and diabetes (Alada, 2000; Dina et al., 2006; Aderibigbe et al.,
1999). Anaemia constitutes a serious health problem in many tropical countries because of
the prevalence of malaria and other parasitic infections. In anaemia there is decreased level
of circulating haemoglobin, less than 13 g dL -1 in male and 12 g dL -1 in females (Okochi et
al., 2003). In the tropics, where malaria is endemic, between 10 to 20% of the population
presents less than 10 g dL -1 of Haemoglobin (Diallo et al., 2008). Children are more
vulnerable. The leaves are rich in iron and play a key role in the cure of anaemia, they are
also noted for lactating properties and are in high demand for nursing mothers ( Okoli and
Mgbeogu., 1983).
Type 2 diabetes is associated with increased oxidative stress, which probably results both
(Baynes, 1991; Tribe and Poston, 1996). In recent years, it has been known that, the most
status, which can induce damage through overproduction of superoxide radical in the
through membranes and initiate lipoperoxidation. The oxidation of unsaturated lipids has
12
implications not only for atherosclerosis, but also for stability and integrity of the red cell
breakdown products like malondialdehyde have been found in erythrocytes and plasma type
glucose level and also have antidiabetic effects in glucose induced hyperglycemic and
streptozotocin (STZ) induced diabetic mice (Aderibigbe et al., 1999), while it did not alter
effects have also been reported by many researchers (Aderibigbe et al., 1999; Eseyin et
produces luxuriant edible green leaves, which are rich in iron and vitamins. Stems of the
plants have branching, long twisting tendrils and the leaves are divided into three to five
leaflets with the terminal leaflets up to 15 cm long, while the male plant is grown
principally for leaves and seeds, which are important soup condition. Recent studies have
pumpkin takes place 120-150 days after sowing ( Longe et al., 1983; Okoli and Nyanayo,
13
al., 2009). The leaves contain essential oils, vitamins; root contains cucubitacine,
sesquiterpene, lactones (Iwu, 1983). The young leaves sliced and mixed with coconut water
and salt are stored in a bottle and used for the treatment of convulsion in ethno medicine
(Gbile, 1986). The leaf extract is useful in the management of cholesterolemia, liver
problems and impaired defense immune systems ( Eseyin et al., 2005a, b).The roots are
used as rodenticide and an ordeal poison ( Gill, 1992). The essential amino acids contents
acid profile of Telfairia occidentalis had also been shown to be very rich and includes
leucine, arginine, serine, threonine, phenylalanine, valine, tyrosine and isoleucine ( Tindall,
1968; Fasuyi, 2006). Emeka and Obidoa (2009) study reveals that the long term feeding
The seeds are highly nutritious and are roasted or boiled and eaten like the seeds of
breadfruit (Treculia); they are also sometimes used as soup thickeners ( Okoli and
Mgbeogu, 1983). The seed is very rich in oil, especially unsaturated fatty acids which form
61% of the oil (Odoemena and Onyeneke, 1998; Okoli and Nyanayo, 1988). Akubue et al.
source of four minerals required in human nutrition. There report showed that the seed
contain 47% oil and 31% protein. The protein was said to be markedly deficient in the
fat, 22% protein, 3% fibre, 15% carbohydrates and 2% ash. Oyolu (1980) observed that
14
vegetables will continue to remain the primary source of proteins, minerals and vitamins in
contain 85% moisture, while the dry portion of what is usually consumed contains
11% crude protein, 25% carbohydrate, 3% oil, 11% ash and as much as 700 ppm iron. The
results of the study carried out by Christian (2007) shows that the seed contained essential
nutrients in significant amount that can supplement other foods. The levels of crude
g -1 ) detected in the seed were compared with nutritional composition of some plant foods in
Nigeria. The study shows that the seed of Telfairia occidentalis Hook F. is high in
carbohydrate, fat and phosphorus. The seed also contained levels of vitamin A which can
supplement other dietary sources. The oil of Telfairia occidentalis seeds have a high iodine
values compared to palm oil which indicates that the oil has a high content of unsaturated
fatty acids relative to palm oil. This suggests that it may be used as edible oil for cooking
The high protein content in leaves of plants such as Telfairia occidentalis could have
supplementary effect for the daily protein requirement of the body. The symptoms of
protein energy malnutrition such as Kwashiorkor and Marasmus were rarely observed
among dwellers in region where adequate amount of protein is obtained from fruits/seeds
and leaves of plants rich in proteins such as Telfaira occidentalis (Dike, 2010; Kayode et
15
Fasuyi and Nonyerem (2007) investigations shows that Telfairia occidentalis leaf meal
hypercholestolemia. This confirms the research work of Eseyin et al. (2005a). The vitamin
investigated by Williams et al. (2009). They observed that Telfairia occidentalis has the
(Williams et al., 2009). The fruits of Telfairia occidentalis have been utilized in the
in medical science. A study carried out by Nwangwu and his colleague shows that Telfairia
occidentalis has the potential to regenerate testicular damage and also increase
Oxygen is used in metabolic reactions. Sometimes it reacts with the metabolic compound to form
a free radical which causes biological damage (Rahman, 2007). Oxygen is a stable product but
when there is gain or loss of an electron in oxygen molecule or its compound it becomes
reactiveoxygen species. Toxicity of oxygen refers to the toxic endpoints of the oxygen
formation.Oxidative stress refers to the distorted balance between the transformation of the reactive
16
oxygenspecies from one form to another indulging beneficial component of cell or
Reactive oxygen species (ROS) play a most important role in variousbiological processes, like as the
oxidative burst reaction which is necessary for phagocytes, andare taking part in a variety of cell
signaling pathways. In the cytosol, IκB is a sequester andinhibitory factor of NF-κB. ROS are also
and NF-κB. So ROS can play a veryimportant role in inflammation modulating. Perturbed reduction
state of cellproduces lipid peroxides, protein carbonyls and other adducts in the cell. These adducts in
celldeposits in clumps to inactivate the normal cell functioning processes of the cell that lead
The reactive oxygen species are the product of cellular metabolism in also normal condition and
readily they are important for cellular signaling pathways. But in some extreme condition,
theaccumulation of ROS up to a limit can cause pathological condition responsible for variousdiseases
(Fanjul-moles, 2016).The reactive oxygen species generated through the process ofoxidative stress are
superoxide radical, hydroxyl radical and hydrogen peroxide. Oxidative DNAdamage is mostly indirect
and adduction of radicals to the DNA can cause mutation so that thecell may become cancerous.
Mitochondrial ETC is one of the types of internal source factor forthe formation of free radicals in
It is a kind of memory loss disease which shatters the memory and other main mental functions.
In this disease, brain cells lose connections, degenerate and finally cells die which cause slowly
17
memory loss. Recent studies reveal that the neuron mitochondria from aging in the rats, are
likeAlzheimer's disease. One of the common forms of dementia thataffect our older population
characterized by degeneration of neurons within the brain. It affectshippocampus region of the brain
which is the responsible part of the brain for memory formationso it causes loss of memory and
. Parkinson’s disease is a common neurodegenerative disease induced bysome stressors from the
endoplasmic reticulum and oxidative damage It is a kind of movement disorder which affects the
central nervous system, responsible for bodymovement so in this disease, patient’s lose body balance
and movements. Over expression of α-synuclein and oxidative stress has been implicated in the
neuronal cell death in the Parkinson’s disease. In Parkinson’s disease, selective degeneration of
It is a nervous system related disease which causes muscle weakness and loss of the movement or
control caused by the motor neurons degeneration. ALS probably leads to a new disease, skeletal
muscledystrophy in infected patients (Kong and Xu,1998). The brain has two types of receptors
glutamate and GABA receptors. Glutamate is achemical mediator which stimulates motor neurons so
its excess formation, is possiblyresponsible for motor neuron destruction in ALS. The GABA receptor
acts to lessen the effectsof glutamate. Gabapentin and Baclofen named medicines act as a GABA
modulators whichincrease GABA activity, are the probable cure for ALS (Amyotrophic lateral
18
Oxidative stress and ROS are most factors of cardiac diseases like congestive heart
failure,hypertension, and atherosclerosis (Sugamura and Keaney, 2011). Smooth muscles and
cardiacmuscles, on the regulation of metabolic process, release free oxygen radicals in the form of
byproducts of the process. This leads to myocardial infarction. Some phagocytescells utilize the LDL
cholesterol and oxidize LDL that lead to the formation of artery cloggingplaque and thus responsible
for the stroke. ROS have been involving inthe important processes which have notable effects on
cardiac functions like hypertrophy, ionflux (calcium), extracellular matrix (ECM) configuration,
metabolism, signaling of variousgrowth factors, cytokines and gene expression. Thioredoxin and
antioxidantand redox regulatory system that has been implicated in a wide variety of ROS-related
antioxidantmolecules, including ubiquinone, lipoic acid, and ascorbic acid, and as such constitute
(Giordano, 2005).
2.2.2.5 Cancer
Free radicals play an important role as the carcinogenic molecule for DNA damage whichinhibits DNA
repair. Attack of free radicals on the hydroxyl group of nucleic acids makes and mutation
fragmentation of nucleic acid thus giving rise to cancer. ROS cause DNA damage andleads to DNA
strand break or fragmentation of the bases. Oxidative products of DNA induceactivation of some
transcription factor like NF-κ B (nuclear factor κ B) that causes abnormal cell
19
growth. Post-transcriptional modification by phosphorylation and nitration by free radicals alsoinduces
cancer risk (Simone et al., 2010). Endogenous and exogenous stimuli participate incancer
on the bases but also the deoxyribosyl backbone of DNA, is one kind of endogenous damage
stimuli which is format through the intermediate oxygen reduction. OFR is a class of important
Remodeling of bone is a natural phenomenon where the bone absorption and formation takes place
during entire life. Less formation of bones and more breaks down will causedeteriorating bone health in
aging people. With the aging of human, could occur the deficienciesof both primary and secondary
Vitamin D which may be participated in the pathogenesis processof senile osteoporosis and also take
part in the development of osteomalacia in an adult humanbeing. Although a mild deficiency could lead
Adarchi, 2011). Decreased mineralcontent and fragility are the symptoms of aging bones. Loss of bone
density may be related toosteoporosis where the bones are more prone to fractures. In the bones, the
rate of repair andregain their mechanical adeptness (power) after fracture, the rate of revascularization,
cellproliferation & differentiation of the cells and bones healing process, decreases with aging(Boskey
Free radical plays significant role in controlling the blood pressure and also fight for the cures
theproduction of free radicals and also its lead to the dysbalance between the production of radicalsand
antioxidants which could lead to oxidative stress with the changes of various biologicalfunctions and
20
structural changes in the cells (Halliwell, 1991). Immune cells participate indefense system by
destroying foreign molecules such as virus or bacteria by producing oxidantsor free radicals. Some
phagocytic cells like macrophages, eosinophils, neutrophils as well as Tand B lymphocyte contain
NADH oxidase which forms superoxide free radicals responsible fordamaging the micro-organisms.
The process is known as the oxidative or respiratory burst. It is the defense mechanism used by the
Antioxidants are the compounds that can stabilize ROS. These molecules are the scavengers offree
radicals and get easily oxidized. Vitamins are the most important class of non-enzymatic antioxidants.
There are two classes water soluble like vitamin C and fat soluble vitamin A(retinoic acid or retinol)
and vitamin E. Vitamin E ( α-tocopherol ) is a predominant scavengerthat has its significant activity in
lipid oxidative damage in both smokers andnonsmokers, so we can conclude that antioxidants vitamins
2.3 Liver
The liver has a particularly intriguing immunological milieu consisting ofa largest reticulo-endothelial
cell network in the body and being a major source of many components of the innate immune response
including acute phase and complement proteins as well as inflammatory cytokines and chemokines.
The organ is also a significant site of immune-mediated damage initiated by infectious, autoimmune
and malignant stimuli. Recent studies have demonstrated that the liver is also an important site of innate
immune system. The innate or natural immune system is the first rapid line-defense against
environmental threats such as microbial infection and physical or chemical injury. Sequential activation
21
of innate and adaptive immune response is crucial for elimination of microorganisms and for immune
response orchestrated by dendritic cells linking innate and adaptive arms of immune system. Unique
repertoires of dendritic and lymphoid cells including NKT cells and regulatory T lymphocytes modify
the immune response in the liver. Non-immune cells of the liver including endothelial cells,
hepatocytes and biliary epithelial cells also contribute to local immunological potential. All of these
elements play roles, together and independently, determining the outcome of immunological
stimulation within the liver. In addition, immune response upon exposure to exogenous or autogenic
agents varies depending on the host genetic backgrounds. The genetic basis of immune response will
offer new approaches to understanding the pathophysiology, diagnosis and management of patients
Liver cells produce the ALT enzyme. ALT levels increase when liver cells are damaged or are dying.
The higher the ALT levels, the more cell death or inflammation of the liver is occurring. However,
ALTs are not always good indications of how well the liver is functioning; only a liver biopsy can
reveal that. ALT levels can remain low even when the liver is inflamed or is developing scar tissue, or
Like the ALT enzyme, AST is an enzyme produced by liver cells, but AST is also produced by muscles
and it can be elevated in conditions other than liver disease. For example, AST is often elevated during
a heart attack. In many cases of liver inflammation, the ALT and AST levels are equally elevated. In
some conditions, such as alcoholic hepatitis, AST levels may be higher than ALT levels. AST levels
22
can be normal, yet there can be liver damage Occurring. This test adds one more perspective to the
Alkaline phosphatase is an enzyme produced in the bile ducts, intestines, kidney, placenta and bone.
This enzyme is measured to help doctors determine if a disease could be concentrated in the bile duct or
in the liver. When this enzyme level is high, and when ALT and AST levels are fairly normal, there
may be a problem in the bile duct, such as an obstruction. Some bone disorders can also cause alkaline
This enzyme, like alkaline phosphatase, is produced in the bile ducts and may become elevated when
there is a bile duct disorder. Elevations in GGT and alkaline phosphatase usually suggest bile duct
disease. Measurement of GGT s an extremely sensitive test; it can be elevated when there is any liver
disease. Elevated GGT levels are also caused by drugs (even when taken at prescribed levels) and they
are often elevated in heavy drinkers, even when there IS no liver disease.
23
CHAPTER THREE
3.0 METHODOLOGY
Telfairia occidentalisleaves and Garlic were collected from Ishara Market, Ishara Remo, Ogun State.
Nigeria.TheTelfairiaoccidentalis leaves were sorted out and washed under running water to
remove dirt. They were chopped into small pieces, oven-dried, and milled into a fine
powder.
The aqueous extract was prepared using Soxhlet extracting method. 65g of the powdered
sample was extracted with 325ml of distilled water.Telfairia occidentalis extraction was done at
the biochemistry laboratory, The Gateway(ICT) polytechnic Sapaade, Ogun state. Nigeria.
3.3.1 APPARATUS
Apparatus used for the experiments are : Soxhlet apparatus, Heating mantle, 250ml beaker, Metal
bead cannula, 2ml syringe, Hand gloves, Distilled water, Whatman filter paper, Nose mask, 5ml
3.3.2 REAGENTS
All reagents used were obtained from recognised dealers and were ensured to be of analytical grade.
Twenty five adult, male Albino rats of approximate age (6-8 weeks) and weight (weighing
200 ± 50 g) were used in the experiment. Animals were kept in clean cages and given
24
normal diet and clean water ad libitum. Animals were acclimated for two weeks before
induction.
These were divided into 5 groups with 5 animals each. The groups are as follow
GROUP TREATMENT
GROUP 1 Normal control Feed + H 2 O
GROUP 2 Negative control 4% Garlic + Feed + H 2 O
GROUP 3 Positive control 20mg/kg Standard drug (Vitamin C) + 4%
Garlic + Feed + H 2 O
GROUP 4 Telfairia occidentalis 200mg/kg extract + 4% Garlic + Feed +
H2O
GROUP 5 Telfairia occidentalis 200mg/kg extract + 4% Garlic + Feed +
H2O
3.6 COLLECTION OF BLOOD SAMPLE
The animals were fasted overnight and blood was collected through cardiac puncture during
the dissecting process. Samples of blood were collected into EDTA bottles and centrifuged
Serum AST, ALT and ALP activities were estimated by the method of Reitman and Frankel, using the
Randox test kit. The activity of alkaline phosphatase (ALP) was assayed using the method of Kochmar
CHAPTER FOUR
4.0 RESULT
25
400 **
***
****
Plasma AST (U/L)
300
***
200
**
*
100
Figu
re 4.1: Aspartate transaminase activity of rats induced with garlic and treated with pumpkin
26
800
Plasma ALP (U/L)
600 ***
***
400 ** **
*
200
Fig
ure 4.2: Alkaline phosphatase activity of rats induced with garlic and treated with pumpkin
27
4.3 Alanine Transaminase Activity
800
***
Plasma ALT (U/L)
600
400 ** **
**
*
200
Fig
ure 4.3: Alanine transaminase activity of rats induced with garlic and treated with pumpkin
28
CHAPTER FIVE
5.1 DISCUSSION
Figure 4.1 showed that the Aspartate transaminase concentration significantly (P>0.05) increased in
rats given garlic only when compared with control. It significantly (P>0.05) decreased in rats given
garlic + pumpkin extracts and garlic + vitamin C when compared with rats given garlic only.
Comparison between rats given pumpkin extracts only and rats given garlic + aqueous pumpkin
extracts showed that the aqueous pumpkin extract significantly reversed the oxidative stress induced by
garlic thereby having a hepatoprotective effect on the rats . This is accompanied by the study of the
long term comsumption of Telfairia occidentalis - supplemented diet by Iweala and Obidoa (2009),
which also showed a non significance of serum protein. This could be associated with the fact that
serum proteins are largely produced in the liver and Oboh (2005) suggested that there could be some
level of effects of Telfairia occidentalis - supplemented diet on rats. The concentration of circulating
triglycerides may be an early and reliable indicator of he patotoxicity in rat and so, its assessment has
been recommend as a part of the core list of liver parameters in preclinical studies. Hepatic damage is
routinely monitored by assaying for specific liver enzyme biomarkers such as AST, ALT and ALP in
Fig 4.2 showed that Alkaline phosphate (P>0.05) increased in rats given garlic only when compared
with control. It significantly (P>0.05) increased in rats given garlic + aqueous pumpkin extract when
compared with rats given garlic + vitamin C. Comparison between rats given aqueous pumpkin extracts
only and rats given garlic + pumpkin extract showed that pumpkin significantly reversed the oxidative
29
stress induced by garlic thereby having a hepatoprotective effect on the rats . This is accompanied by
the study of the long term consumption of Telfairia occidentalis - supplemented diet by Iweala and
Obidoa (2009), which also showed a non significance of serum protein. This could be associated with
the fact that serum proteins are largely produced in the liver and Oboh (2005) suggested that there
could be some level of hepatoprotective effects of Telfairia occidentalis - supplemented diet on rats.
The concentration of circulating triglycerides may be an early and reliable indicator of hepatotoxicity in
rat and so, its assessment has been recommend as a part of the core list of liver parameters in preclinical
studies. Hepatic damage is routinely monitored by assaying for specific liver enzyme biomarkers such
as AST, ALT and ALP in the liver and serum of experimental animals (Shivaraj et al., 2009).
Figure 4.3 showed that the Alanine Transaminase concentration significantly (P>0.05) increased in rats
given garlic only when compared with control. It significantly (P>0.05) decreased in rats given garlic +
pumpkin extracts and garlic + vitamin C when compared with rats given garlic only. Comparison
between rats given pumpkin extracts only and rats given garlic + pumpkin extracts showed that the
aqueous pumpkin extract significantly reversed the oxidative stress induced by garlic thereby having a
hepatoprotective effect on the rats. This is accompanied by the study of the long term consumption of
Telfairia occidentalis - supplemented diet by Iweala and Obidoa (2009), which also showed a non
significance of serum protein. This could be associated with the fact that serum proteins are largely
produced in the liver and Oboh (2005) suggested that there could be some level of hepatoprotective
triglycerides may be an early and reliable indicator of hepatotoxicity in rat and so, its assessment has
been recommend as a part of the core list of liver parameters in preclinical studies. Hepatic damage is
routinely monitored by assaying for specific liver enzyme biomarkers such as AST, ALT and ALP in
30
5.2 CONCLUSION
The medicinal potential of the leaf and of Telfairia occidentalis is no longer in doubt. It is now obvious
that the plant has been proven topossess beneficial antioxidant, antidiabetic, hepatoprotective,
anxiolytic and sedative properties. Many of these activities confirm the claims of herbal medical
practitioners, although clinical tests, using human subject, still need to be done. Many researchers have
also attributed some of the medicinal activities of the plant to the high level of antioxidant components
(such asphenolics, flavonoids, Vitamins C and E) and antioxidant property of the plant. However, the
presence of other phytochemicals such as saponins may also play valuable roles in the activities of the
plant.
31
REFRENCES
of Telfairia occidentalis (fluted pumpkin) in rats fed a cholesterol-rich diet. J. Med. Food, 10:
330-336.
Ajibade, S.R., Balogun, M.O., Afolabi, O.O.and Kupolati, M.D., (2006). Sex differences in
Nigeria.
Akoroda, M.O., (1990). Seed production and breeding potential of the fluted pumpkin, Telfairia
occidentalis.
(1990). Flowering, pollination and fruiting in fluted pumpkin (Telfairia occidentalis). 43: 197-
206.
Akubue, P.I.,Kar, A. and Nnachetta, F.N., (1980). Toxicity of extracts of roots and leaves
Asiegbu, I.E., (1987). Some biochemical evaluation of fluted pumpkin seed. J. Sci. Food Agric., 40:
151-155.
32
Badifu, G.I.O. and Ogunsua,A.O., (1991). Chemical Composition of kernels from some speciesof
40: 405-412.
Brownlee, M., (2001). Biochemistry and molecular cell biology of diabetic complications. Nature,
414: 813-820
Cowan, M.M., (1999). Plant products as antimicrobial agents. Clin. Microbiol. Rev., 12: 564-582.
Dike, M.C., (2010). Proximate, phytochemical and nutrient compositions of some fruits, seeds and
leaves of some plant species at umudike, Nigeria. ARPN J. Agric. Biol. Sci., 5: 7-16
Res., 3: 181-183.
Egbekun, M.K., Nda-Suleiman E.O., and Akinyeye, O., (1998). Utilization of fluted pumpkin fruit
(Telfairia occidentalis) in marmalade manufacturing. Plant Food Hum. Nutr., 52: 171-176.
Emeka, E.J.I.and Obidoa, O., (2009). Some biochemical, haematological and histological responses
1199-1203.
33
Eseyin, A.O.,Igboasoiyi, C.A., Mbagwu, H., Umoh, E.and Ekpe, F J., (2005). Studies on the effects
of an alcohol extract of the leaves of Telfairia occidentialis on alloxan induced diabetic rats.
seed extract of Telfairia occidentalis in rat. Pak. J. Biol. Sci., 10: 498-501.
Eseyin, O.A., Ekpo, A., Idem, I., Igboasoiyi, A.C. and Edoho, E.J., (2007). Effects of the fruit
of Telfairia occidentalis on some biomolecules in rat. Pak. J. Biol. Sci., 10: 3240-3242.
of Telfairia occidental is leaves on some biochemical parameters in rat. Global J. Pure Applied
Eseyin, O.A.,Oforah, E.and Dooka, B.D., (2000). Preliminary study of the hypoglycemic action of
the extract of leaf of Telfairea occidentalis in normoglycemic guinea pigs. Global J. Pure
Biotechnol., 6: 1055-1063.
Fasuyi, A.O., (2006). Nutritional potentials of some tropical vegetable leaf meals: Chemical
Gbile, Z.O., (1986). Ethnobotany, Taxonomy and Conservation of Medicinal Plants. In: The State of
Medicinal Plants Research in Nigeria, Sofowora, A. (Ed.). University of Ibadan Press, Ibadan,
Nigeria.
34
Gill, L.S., (1992). Ethromedical Uses of Plants in Nigeria. Uniben Press, University of Benin, Benin
1st Edn., Chapman and Hall, London, UK., ISBN: 978-94-009-5921-7, Pages: 271.
Horsfall, Jr.M.andSpiff, A.I., (2005). Equilibrium sorption study of Al3+, Co2+ and Ag+ in aqueous
solutions by fluted pumpkin (Telfairia occidentalis Hook f) waste biomass. Acta Chim. Slov.,
52: 174-181.
Iwu, M.M.,Duncan A.R. and Okunji, C.O., (1999). New Antimicrobials of Plant Origin. In:
Perspectives on New Crops and New Uses, Janick, J. (Ed.). ASHS Press, Alexandria, VA.,
Iwu, M.W., (1983). Traditional Igbo Medicine. Institute of African Studies University of Nigeria,
Nsukka..
Kayode, O.T., Kayode A.A. and Odetola, A.A., (2009). Therapeutic effect of Telfairia occidentalis
Ladeji, O.,Okoye, Z.S.C. and Ojobe,T., (1995). Chemical evaluation of the nutritive value of leaf of
the extract of Telfairia occidentalis in rats. Caderno Pesquisa Biologia, 19: 27-35.
35
Nwanna, E.E. and Oboh, G., (2007). Antioxidantand hepatoprotective properties of polyphenol
Nwozo, S.O.,Adaramoye, O.A. and Ajaiyeoba, E.O., (2004). Anti-diabetic and hypolipidemic studies
of Telifairia occidentalis on alloxan induced diabetic rabbits. Niger. J. Natl. Prod. Med., 8: 45-
47.
Oboh, G. andAkindahunsi, A.A., (2004). Change in the ascorbic acid, total phenol and antioxidant
activity of sun-dried commonly consumed green leafy vegetables in Nigeria. Nutr. Health, 18:
29-36.
Oboh, G., (2005). Hepatoprotective property of ethanolic and aqueous extracts of fluted pumpkin
(Telfairia occidentalis) leaves against garlic-induced oxidative stress. J. Med. Food, 8: 560-
563.
Odoemena, C.S. andEssien, J.P., (1995). Antibacterial activity of the root extract of Telfairia
Odoemena, C.S. andOnyeneke, E.C., (1998). Lipids of fluted pumpkin (Telfairia occidentalis) seeds.
Proceedings of the 1st African Conference on Biochemistry of Lipids, September 13-16, 1998,
Okochi, V.I., Okpuzor, J. andAlli, L.A., (2003). Comparison of an African herbal formula with
Okokon, J.E., Ekpo, A.J. andEseyin, O.A., (2007). Antiplasmodial activity of ethanolic root extract
36
Okoli, B.E. and C.M. Mgbeogu, (1983). Fluted pumpkin, Telfairia occidentalis: West African
Oyolu, C., (1980). Maximizing the contribution of Okra (Hibiscus esculentus) to the national diet.
Proceedings of the 3rd Conference Host Society, November 30-December 3, 1980, Nigeria at
University of Ife -.
occidentalis leaves reduces blood sugar and increases haematological and reproductive indices
Shah, J.S.,Shah, M.B., Goswami, S.S. and Santani, D.D., (2006). Mechanism of action of antiulcer
Sofowora, A., (1993). Medicinal Plants and Traditional Medicine in Africa. 2nd Edn., Spectrum
Tribe, R.M. andPoston, L., (1996). Oxidative stress and lipids in diabetes: A role in endothelium
medicine. WHO Document No. WHA30.49, World Health Organization, Geneva, Switzerland.
37
WHO., (2001). Geneva Legal Status of Traditional Medicine and Complementary/Alternative
Williams, I.O.,Parker, R.S. and Swanson, J., (2009). Vitamin a content of southeastern nigerian
Zheng, W. andWang, S.Y., (2001). Antioxidant activity and phenolic compounds in selected herbs. J.
38