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ABSTRACT
Title:
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2015-2016
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This study was conducted to evaluate a wound healing rate of Buddha belly leaf
(Jatropha podagrica) ointment on incised wound of albino rats.
30 albino rats were randomly distributed into five treatment groups with 6
replicate each group. The Buddha belly leaf extract (1ml, 1.5ml and 2ml) was
incorporated into 10 g of a simple ointment base by melting and trituration to produce
three batches of the ointment formulation. Incision wound measuring 1 inch (2.5 cm) was
created on the dorsal pelvic region of the albino rats and the ointment was applied
topically twice a day on the wound which was measured at intervals of 3 days until
epithelialization or complete wound closure. The other group was treated with Bactroban
ointment (2%) which serve as the control and standard treatments, respectively. And the
last group were remained untreated and heals naturally.
Topical application of the leaf extract of Jatropha podagrica incorporated into an
ointment base on the incision wound in rats caused a significantly (P<0.05) higher rate of
wound healing and reduced the epithelialization period in a doserelated manner. The
result shows that the ointment formulated from extracts of Jatropha podagrica containing
the highest concentration of the sample extract (2ml/10g ointment) showed the highest
rate of wound closure when compared to the no applied ointment treated group as well as
the formulated ointment containing the lower concentrations (1ml and 1.5ml/10g of
ointment base).
INTRODUCTION
Topical antimicrobial therapy is one of the most important methods of wound
care. Some medicinal plants have been employed in folk medicine for wound. Some of
these plants either possess pro-wound healing activities or exhibit antimicrobial and other
related properties which are beneficial in overall wound care. Wound healing processes
are well organized biochemical and cellular events leading to the growth and regeneration
of wounded tissue in a special manner. Healing of wounds is an important biological
process involving tissue repairs and regeneration. It involves the activity of an intricate
network of blood cells, cytokines, and growth factors which ultimately leads to the
restoration to normal condition of the injured skin or tissue (Clark, 1991).
Since the earliest times, people rely on herbs for treatment of various deseases.
Herbs became an affective source of human health due to the active compounds present
within rendering them many pharmacological activities. Treatment may not heal as fast as
commercial drugs that we have now but they proved to be cheap and efficient (Longid
2014). One of the frequently used herbs is the Buddha belly plant (Jatropha podagrica).
The genus name Jatropha derives from the Greek word jatros (doctor/healer) and
trophe (food), which implies medicinal uses and the species podagrica (podagrikos) as a
liable to gout. Jatropha podagrica Hook (various names: Buddha belly, Gout plant, Coral
nut, Guatemala rhubarb, physic nut; Family Euphorbiaceae) is a shrub native to tropical
including
antimicrobial,
antibacterial,
antitumour,
anti-inflammatory,
molluscidal and insect antifeedant have been attributed to this plant. It is also used as an
antipyretic, diuretic, choleretic, purgative, reduction of swelling, pain relief and to
detoxify snakebites and treatment of scabies.
Those qualities claimed present in Buddha belly are the reason for the researcher
to investigate a methanol extract of Jatropha podagrica (Euphorbiaceae) which was
formulated in an ointment base for pro-wound healing activity on incision wound model.
Definition of terms:
Antimicrobial: inhibiting the growth of microorganisms.
Chapter II
REVIEW OF LITERATURE
Traditional Use:
Traditionally this plant is considered as being sweet, bitter and cold. It has been
used as analgesic, antipyretic, ant-inflammatory and detoxicant (Globinmed).
In Indonesia the whole plant is used to treat fever. The Africans however make
use of the seed oil as part of the ingredients in a compound medicine to treat fever. It has
been used in the treatment of malaria. The whole plant is used as haemostatic, and also
applied on hematoma to aid in rapid reduction of swelling. The plant is known to have
diuretic properties and is used to treat haematuria. It is also used to treat gonorrhea. In
Indonesia and china, the plant has been used to treat snake bites 10-15g of the plant is
pounded and then immersed in rice wine (Globinmed).
Schistosomicidal Activity
An organization known as Global Information Hub on Integrated Medicine, also
said that Tetramethylpyrazine (TMP) isolated from the stem bark of Jatropha podagrica
Cytotoxic activity
It was also shown that the fraxidin, acetylaleuritolic acid and g-sitosterol isolated
from roots and stem showed cytotoxic activities against HeLa cell lines with IC 50 of
39.9ug/L, 35.7ug/L and 15.9ug/L respectively (Globinmed).
A previous study from the University of Rouen, Mont St. Aignan entitled
Assessment of cyto-protective, antiproliferative and antioxidant potential of a
medicinal plant Jatropha podagrica that aimed to evaluate antiproliferative,
antioxidant activity and in vitro protective effect of hydro-alcoholic extract of Jatropha
podagrica. Total phenolics and flavonoids content was higher in seeds than leaves
extract. One of their new journalist obtained a quote from the research states that: The
anti-proliferative effect on two tumoral celllines A549 and PC12 was assessed by MTT
assay. The cytotoxicity of the studied extract was evaluated on young cerebellar granule
cells. The investigation of the action mode off. Podagrica extract in protection of DNA
damages, protein carbonylation and lipid peroxidation caused by hydroxyl radicals
generated in Fentons reaction exhibits an effective protection at low concentrations. In
the presence of 100 p,M of copper sulfate, Jatropha hydro-alcoholic extract showed no
significant enhancing in the hydroxyl radicals generation and DNA fragmentation in
vitro.
In anesthesize cats it produced depressor effects, reduced the heart rate and
blocked neuromuscular transmission and appeared to have spasmolytic activity on
smooth muscles (Ojewole, 1980). Further studies confirmed blockage of adrenergic and
cholinergic transmission by tetramethylpyrazine. The compound depressed and abolished
the electrically evoked contractions of the chick oesophagus, rabbit duodenum and guinea
pig vas deferens in vitro. It is also inhibited the electrically induced contraction of the rat
isolated hemi-diaphragm and of the cats nictitating membrane in vivo. Apart from its
possible central effects, and those on the cardiac muscle and blood vessels, it could be
suggested, from the results obtained in the study, that the hypotensive effect in
experimental animals is likely to be contributed to by, or mediated via, its local anesthetic
(membrane stabilizing) activity. Through this action, the drug probably acts to block
sympathetic and parasymphatetic neurons and ganglia (Ojewole, 1981).
Furthermore, tetramethylpyrazine has a number of other pharmacological actions.
A main central effect was found to be tranquillization and sedation (Ojewole and Odebiji,
1984).
In china, tetramethylpyrazine originating from plants is used in the treatment of
occlusive cerebral vessel diseases such as cerebral embolism (Xiao, 1983).
Synthesis of Collagen
1. Combination of amino acid to form chains
2. Chains associate to form molecules
3. Molecules associate to form fibrils
4. Fibrils aggregate into fibers or bundles
Laboratory Animals
The Laboratory rats like its wild counterpart is an extremely intelligent animal,
probably more so than the other rodents commonly used for biomedical research.
Scientists and researchers rely on mice and rats for several reasons. One is convenience:
rodents are small, easily housed and maintained, and adapt well to new surroundings.
They also reproduce quickly and have a short lifespan of two to three years. Rats are also
relatively inexpensive and can be bought in large quantities from commercial producers
that breed rodents specifically for research. The rodents are also generally mild-tempered
and docile, making them easy for researchers to handle (Robert W. Kemp).
Another reason rodents are used as models in medical testing is that their genetic,
biological and behavior characteristics closely resemble those of humans, and many
symptoms of human conditions can be replicated in mice and rats. "Rats and mice are
mammals that share many processes with humans and are appropriate for use to answer
many research questions," said Jenny Haliski, a representative for the National Institutes
of Health (NIH) Office of Laboratory Animal Welfare.
METHODOLOGY
Materials
The materials used in this study are: basin, knife, chopping board, blender,
beakers, weighing balance, whatman no.2 filter paper, graduated cylinder, spatula,
stirring rod, water bath, surgical blade, cotton, tissue, forceps, syringe, and caliper, as
well as the Albino rats and the Buddha belly leaves (Jatropha podagrica).
Chemicals
The drug that was used for the comparison of commercial drug from the Buddha
belly leaves extract is Bactroban (2%).
Treatments
T1 = 1ml/10g of ointment base
T2 = 1.5ml/10g of ointment base
T3 = 2ml/10g of ointment base
T4 = Bactroban ointment 2% (Positive control)
T5 = No Ointment applied (Negative control)
Induction of Wounds
The predetermine area for wound induction at the back of the animal were
prepared for surgery by removing the dorsal fur or hairs with a pair of scissors and
scalpel blade. The animals were held in standard crouching position, and the mobile skin
were gently stretched and held by fingers. The induction of a tranquilizer drug
Acepromazine maleate (.1ml/250g of rat) were done by IM injection and the induction of
localized anesthesia were also done by subcutaneous injection of a lidocaine solution
(0.3ml/2%) at the dorsal part of the animal and around the area under investigation to
render area painless. After five(5) to eight(8) minutes of introducing local anesthesia, the
rat were then subjected for incisional wound (Celestino et al, 2014).
The anticipated area of the wound created were outlined on the back of the
animals. A longitudinal paralumbar incision, 1 inch in length and .2 cm in
depth were incised through the skin using toothed forceps, a surgical blade
and pointed scissors. After achieving complete homeostasis by blotting the wound with
cotton swab soak in warm saline, the animals were placed singly in individual cages.
Statistical analysis
The date were analyzed using one way analysis of variance (ANOVA and data
subjected to LSD post hoc test. Differences in mean between paired observations were
accepted as significant at P 0.05. Also, the data will be analyzed using tables, means, and
percentages.
LITERATURE CITED