ABSTRACT
Combretum racemosum (P. Beauv.) commonly known as “Christmas rose” in Southern Nigeria local English is a plant that has been in use for several years by African traditional healers in the treatment of haemorrhoids, convulsive coughing, tuberculosis, toothache, male sterility and as condiments in soups. This study was designed to evaluate the in vitro antioxidant and diuretic activities of the ethanolic extract of C. racemosum leaves on experimental model albino rats. The acute toxicity (LD50) of C. racemosum leaves was carried out on the albino rats using standard method. The result showed a lethal dose (LD50) > 5500mg/kg of the extract. In the in vitro antioxidant test, there was a concentration dependent increase in the percentage (%) scavenging activity when compared with the Vitamin C (control) for 2,2-diphenyl-1-picrylhydrazyl (DPPH), Nitric oxide and Anti-lipid peroxidation activity of C. racemosum leaves. The result exhibited a significant difference at higher concentration when compared to the control. The IC50 values of the plant extract were observed which measures its antioxidant effectiveness. The ethanolic extract also exhibited a significant increase in urine volume and the electrolyte at different concentration from 187.5mg/ml to 3000mg/ml. At 3000mg/ml (0.7 ± 0.14) and 1500mg/ml (0.45 ± 0.07) concentrations, the extract exhibited a significant difference in sodium (Na+) excretion (87.4 ± 1.70) and (76.65 ± 0.49) respectively. The potassium (K+), chloride (Cl-) and bicarbonate (HCO3-) level decreases as the concentration decreases from 3000mg/ml to 187.5mg/ml. The decrease in the electrolyte level as the concentration of the doses decreases exhibited a significant difference (P<0.05). From the result generated, the ethanolic extract of C. racemosum leaves is an effective natural antioxidant and can induce diuresis at very high dosage.
TABLE OF CONTENT
Title page i
Declaration ii
Certification iii
Dedication iv
Acknowledgement v
Table of contents vi
List of tables’ viii
List of plates and figures ix
Abstract x
CHAPTER
ONE
1.0
Introduction 1
1.1 background
of study 1
1.2 Aims
and objectives 3
1.3 Justification 3
CHAPTER
TWO
2.0 Literature
review 4
2.1 Medicinal
plants 4
2.2 Botanical
description of Combretum racemosum 5
2.2.1 Uses
of Combretum racemosum 6
2.3 Antioxidant 6
2.4 Diuretics 9
CHAPTER
THREE
3.0
Materials and Methods 14
3.1 Material 14
3.1.1 Plant
material 14
3.1.2 Extraction
of the plant material 14
3.1.3 Experimental
animal 14
3.2 Antioxidant
activity of Combretum racemosum leaves 15
3.2.1 Determination
of DPPH radical scavenging activity 16
3.2.2
Determination
of Nitric oxide scavenging activity 16
3.2.3 Determination
of Anti-lipid peroxidation activity 16
3.3 Diuretic
activity 17
3.3.1 Experimental
design 17
3.3.2 Measurement
of urine output and analysis of electrolyte 17
3.4 Lethal
Dose (LD50) 18
3.4.1 Acute
toxicity test 18
3.5 Statistical
analysis 18
CHAPTER
FOUR
4.0 Result 20
4.1 Acute
toxicity result 20
4.2 Evaluation
of ethanolic extract fraction of C.
racemosum for antioxidant activity 20
4.2.1 DPPH
photometric assay 20
4.2.2 Evaluation
of Nitric oxide inhibition activity of C.
racemosum and Vitamin C 21
4.2.3 Evaluation
of lipid peroxidation activity of C.
racemosum and Vitamin C 22
4.3 Diuretic
effect 23
CHAPTER FIVE
5.0 Discussion 28
5.1 Conclusion 31
REFERENCES
LIST OF TABLES
TABLE 1: Result
of acute toxicity test
TABLE 2: Result
of 2,2-diphenyl-1-picrylhydrazyl (DPPH) with Vitamin C
TABLE 3: Result
of Nitric Oxide inhibition activity with Vitamin C
TABLE 4: Result
of Anti-lipid peroxidation activity with Vitamin C
TABLE 5: Diuretic
result
LIST OF PLATES AND FIGURES
PLATE 1: Experimental
animal
FIGURE 1: Combretum racemosum
FIGURE
2: Bar chart showing the different level
of urine volume
CHAPTER
ONE
INTRODUCTION
1.1 BACKGROUND OF STUDY
The Combretaceae family
includes more than 600 species distributed among about 20 genera, with
pantropical distribution. Combretum
is one of the two most commonly occurring genera with about 370 species and is
widely used in Africa (Stace, 2007). Species of Combretum have been of interest in the last two decades due to the
isolation of some compounds with highly significant activities in anticancer
anti-infectious model (McKeage, 2011; Okwuosa et al., 2006). This has made the Combretum a very important group to search for bioactive compounds.
Combretum racemosum
(P. Beauv.) is a straggling shrub
widespread across Africa that bears a mass of crimson flowers which is very
spectacular, commonly known as Christmas rose in the Southern Nigerian local
English (Burkill, 1985). The plant has been used for several years in African
traditional medical practices and as a condiment in soups. In addition to its
anthelmintics (Okwuosa et al., 2006),
trypanocidal and antimicrobial properties for genitor-urinary and
gastrointestinal infections (Onocha et
al., 2005), the plant is also used for the treatment of haemorrhoids,
convulsive coughing, tuberculosis, toothache and male sterility (Burkill, 1985).
Antioxidants are naturally
occurring plant substances that protect the body from damage caused by harmful
molecules called free radicals. Antioxidants help prevent oxidation, which can
cause damage to cells and may contribute to aging. They may improve immune
functions and perhaps lower the risk for infections, cardiovascular diseases
and cancer. Antioxidant exists as vitamins, minerals and other compounds in
foods. A diet containing plenty of fruits and vegetables, whole grains and nuts
can supply all the antioxidants your body needs. Diets rich in antioxidants can
be very beneficial. Antioxidant is a molecule capable of inhibiting the
oxidation of other molecules. Oxidation is a chemical reaction that transfers
electrons from a substance to an oxidizing agent. Oxidation reaction can
produce free radicals. In turns this radicals can start chain reactions that
damage cells. Antioxidants terminate these chain reactions by removing free
radical intermediates, and inhibit other oxidation reactions. They do this by
being oxidized themselves, so antioxidants are often reducing agents such as
thiols, ascorbic acid or polyphenols (Sies, 1997). Although oxidation reactions
are crucial for life, they can also be damaging; hence plants and animals
maintain complex systems of multiple types of antioxidants such as glutathione,
vitamin C, and vitamin E as well as enzymes such as catalase, superoxide
dismutase and various peroxidases. Low levels of antioxidants or inhibition of
the antioxidant enzymes, causes oxidative stress and may damage or kill cells.
Antioxidants are widely used as ingredients in dietary supplements in the hope
of maintaining health and preventing diseases such as cancer and coronary heart
disease.
Although initial studies
suggested that antioxidant supplements might promote health, later large
clinical trials did not detect any benefit and suggested instead that excess
supplementation may be harmful (Bjelakovic et
al., 2007). In addition to these uses of natural antioxidants in medicine,
these compounds have many industrial uses such as preservatives in food and
cosmetics and preventing the degradation of rubber and gasoline.
Diuretic are commonly defined
as drugs that increase the amount of urine output by the kidneys. One of the
important and well documented uses of plant products is their use as diuretic
agents. These agents augment the renal excretion of sodium and either chloride
or bicarbonate primarily, and water excretion secondarily (Barrar, 2003). Diuretic
drugs increase the rate of urine flow and sodium excretion, and are used to
adjust the volume and composition of body fluids in a variety of clinical
situations. Drug-induced diuresis is helpful in many
life-threatening conditions such as congestive cardiac failure (CCF), nephritic
syndrome, cirrhosis, renal failure, toxemia of pregnancy, premenstrual tension,
and hypertension (Pandya et al.,
2012; Sravani et al., 2010). Most
diuretic drugs have the adverse effect on quality of life, inducing impotence,
fatigue and weakness. Naturally occurring diuretics include caffeine, coffee tea, and Kola accuminata which inhibit sodium
reabsorption of alcoholic beer (Umesh et
al., 2010). Most indigenous drugs have been claimed to have diuretic
effects.
1.2 AIM AND OBJECTIVE
The aim and objective of this
research is to determine the antioxidant and diuretic activities of the ethanolic
extract of the leaves of C. racemosum on
wister albino rats.
1.3 JUSTIFICATION
This study was undertaken to
justify the claims of traditional healers that C. racemosum can be used
to treat oxidative stress caused by free radicals and induce diuretic
activities and to also know if other benefits can be derived from the plant
which can be utilized in the chemical pharmaceutical industries.
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