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EVALUATION OF THE EFFECTS OF GONGRONEMA LATIFOLIUM ETHANOL LEAF

EXTRACT ON THE LIVER ENZYMES OF IN THE ALLOXAN- INDUCED DIABETIC
ALBINO RATS

Article · July 2022

DOI: 10.46344/JBINO.2022.v11i03.41

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J.Bio.Innov11(3), pp: 990-996, 2022 |ISSN 2277-8330 (Electronic) Obeagu et al.,

https://doi.org/10.46344/JBINO.2022.v11i03.41

EVALUATION OF THE EFFECTS OF GONGRONEMA LATIFOLIUM ETHANOL LEAF EXTRACT ON THE

LIVER ENZYMES OF IN THE ALLOXAN- INDUCED DIABETIC ALBINO RATS

Joseph Chikezie Enye1, *Emmanuel Ifeanyi Obeagu2,3,Richard I. Eze3 and P.E. Eze-Steven

1Department of Pharmacy, Madonna University, Nigeria, Elele Campus, Nigeria.

2Department of Medical Laboratory Science, Imo State University, Owerri, Nigeria

3Department of Medical Laboratory Science, Madonna University, Nigeria

4Department of Applied Biochemistry, Enugu State University of Science and Technology, Enugu State, Nigeria

emmanuelobeagu@yahoo.com

ABSTRACT
Gongrenema latifolium is used by traditional healers in different localities for the treatment
of various ailments including Diabetes mellitus without any scientific proof or validation. This
study is aimed at investigating the effects of ethanol leaf extracts of Gongrenema
latifolium on liver enzymes in adult albino rats. Statistical analysis was carried out using
ANOVA. The different doses of the extracts also showed a significant decrease (p<0.05) in
the value of Alkaline phosphatase (ALP), Alanin aminotransferase (ALT) and Aspartate
aminotransferase (ASP). The extracts did not show any adverse effect in the liver based on
the liver function test function test. This study has therefore provided some vital pieces of
scientific information about the hepatoprotective effects of the extracts of Gongronema
latifolium which can possibly contribute to its use as a herbal therapy in the management
of diabetic patients

Keywords: Gongronema latifolium, liver enzymes, Alloxan- induced diabetic albino rats

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J.Bio.Innov11(3), pp: 990-996, 2022 |ISSN 2277-8330 (Electronic) Obeagu et al.,

INTRODUCTION Herbalism has a long tradition of use

The use of plants for healing purposes outside of conventional medicine. It is
predates human Herbal medicine, also becoming more main stream as
called botanical medicine or improvements in analysis and quality
phytomedicine refers to the use of parts of control along with advances in clinical
plants such as seeds, berries, roots, leaves, research show the value of herbal
bark or flowers for medicinal purposes medicine in treating and preventing of
(Ehrlich, 2019). It may also be defined as disease (Ehrlich, 2019).
popular stock or knowledge about Blood glucose levels are controlled by a
medicinal properties of herbs and roots as complex interaction of multiple chemicals
treatment for common remedies and other and hormones in the body including the
diseases in the society, which had been hormone insulin made in the beta cells of
handed down from generation to the pancreas. Diabetes mellitus consists of
generation (Ayodhya and Baba, 2017). a group of syndromes characterized by
Studies have confirmed the benefits of hyperglycaemia, altered metabolism of
medicinal plants with hypoglycaemic lipids, carbohydrates, and proteins; and an
effects in the management of diabetes increased risk of complications from
mellitus (Bnouham et al, 2016). Numerous vascular disease. Criteria for the diagnosis
mechanisms of actions have been of diabetes mellitus have been proposed
proposed for these plant extracts. Some by several medical organizations (WHO,
hypotheses relate to their effects on the 2919). In recent years, developed and
activity of pancreatic B-cells (synthesis, developing nations have witnessed an
release, and cell regeneration) or the explosive increase in the prevalence of
increase in the protective/inhibitory effect diabetes mellitus predominately related to
against insulinase and the increase in life style changes and the resulting surge in
insulin sensitivity or the insulin-like activity of obesity (King et al, 2018). The metabolic
the plant extracts. Other mechanisms may consequences of prolonged
involve improved glucose homeostasis hyperglycemia and dyslipidemia, including
(increase in peripheral utilization of accelerated atherosclerosis, chronic
glucose, increase in synthesis of hepatic kidney disease and blindness pose an
glycogen and/or inhibition of intestinal enormous burden on patients with DM and
glucose absorption), reduction of on the public health system (Goodman
glycaemic index of carbohydrates, and Gilman, 2016).
reduction of the effect of glutathione. All Diabetes mellitus occurs throughout the
of these actions may be responsible for the world but is more common (especially type
reduction and or abolition of diabetes 2) in the more developed countries
complication (Bnouham et al, 2016). (Rother, 2017). The greatest increase on

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prevalence is, however, expected to in the animal house under room

occur in Asia and Africa, where most temperature and were allowed to
patients will likely be found by 2030 (Roglic acclimatize for two weeks before the
et al, 2014). The increase in incidence of experiment. The animals were fed on
diabetes in developing countries follows commercial pellet feeds and were given
the trend of urbanization and lifestyle potable water ad libitum. They were fasted
changes, perhaps most importantly a for 12hours before the commencement of
"western-style" diet. For past 20 years, the experiment, but were allowed water
diabetes rates in North America have ad libitum.
been increasing substantially. Animal Ethics Approval from University
The study was done to determine the senate Research committee
effect of the Gongronema latifolium The study was conducted after obtaining
ethanol leaf extract on the liver enzymes full approval (reference number:
such as aspartate amino-transferase (AST), MAU/SREC/A/17) from University senate
Alanine amino transferase (ALT) and Research and ethics committee of
alkaline phosephatase (ALP) in the Alloxan- Madonna university Nigeria
induced diabetic albino rats. EXTRACTION OF PLANT EXTRACT MATERIALS
MATERIALS AND METHODS The leaves of Gongronema latiflium were
STUDY AREA thoroughly washed with clean tap water
This study was carried out in the and air-dried in the laboratory for 14 days.
department of Pharmacology, faculty of The leaves were pulverized into fine
Pharmacy,Madonna University, Elele Rivers powder using electric grinding machine.
State, Nigeria. Three vehicles were used for the
Collection and identification of plant extraction. For ethanol extraction, 500g of
The leaves of Gongonema latifolium plant the powdered material was soaked in
were collected from a farmland in Umunze 1000ml of 80% ethanol, extract was
in Anambra State, South East, Nigeria and concentrated using a rotary evaporator.
were identified by Mrs.Victoria Alozie, a The same process was repeated using
taxonomist in Botany department of distilled water and methanol respectively.
University of Nigeria Nsukka where a The solid ethanol extract of the ethanol
voucher specimen was deposited in the was kept in air-tight container and stored in
herbarium a refrigerator and was used throughout the
EXPERIMENTAL ANIMALS study of 40%.
One hundred and thirteen (113) adult DETERMINATION OF YIELD OF PLANT
albino rats of both sexes weighing (150- EXTRACT
180)g were obtained from the animal This was carried out according to method
house of the department of Pharmacology proposed to Okoli et al, (2010). The plant
and toxicology Madonna University, Elele, extract was evaporated to dryness at a
Rivers State Nigeria. The animals were temperature of 40• in a hot oven using a
housed in cages in a well-ventilated area previously weighed empty beaker.
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Yield in percent was calculated using the glucose enzymatic-colorimetric test kit-
formula; Glucose oxidase-peroxidase (GOD-POD)
Yield weight (g) of residue x 100 method, produced by Cyprus diagnostics
-------------------------------------------- (Belgium). The test principle is based on the
Weight of coarse powder oxidation of glucose by glucose oxidase
(GOD) to gluconic acid and hydrogen
Induction of hyperglycaemia peroxide. The hydrogen
(experimental diabetes) peroxide(H202)forms a red violet colour
Alloxan (Sigma St. Louis U. S. A.) was with a chromogenic oxygen acceptor,
dissolved in normal saline and was given to phenolaminophenazone in the presence
the animals at a dose of 150mg/kg of peroxidase (POD). The colour intensity is
intraperitoneally (LP) for 7days. Rats with proportional to glucose concentration in
blood glucose levels equal to or greater the sample and the results were recorded
than 200mg/dl (11.lmmol/ were considered as mMol/1.
diabetic and used for this study. Estimation of Liver Enzymes in the alloxan-
Anti-diabetic treatment (Experimental induced Albino Rats after treatment
Design) withEthanol extract of Gongronema
Alloxan-induced diabetic albino rats which latifolium (Tietz, 2018)
were fasted for 12 hrs were placed in five After the alloxan-induced diabetic albino
(5) groups (1-5) of 20 rats each and treated rats were treated for 14 days using the
as follows; Group 1: Received normal saline Ethanol leaf extract of Gongronema
intraperitoneally (ip) (negative). latifolium, the treated albino rats were
Group 2: Received 5mg/kg body weight of placed into three (3) groups of twenty rats
glibenelamide intraperitoneally (i.p.) per group. Group 1 that received
(positive control) 250mg/kg body weight, group 2 that
Group 3: Received 250mg/kg body weight received 500mg/kg body weight of the
of the ethanol leaf extract ip. Group 4: Ethanol extract and group 3 that received
Received 500mg/kg body weight of the l000mg/kg body weight of the Ethanol
ethanol leaf extract ip. Group 5: Received extract. Blood samples for Liver Enzyme
l000mg/kg body weight of the ethanol leaf assay were collected in tubes by cutting
extract ip. All these groups received these the tail-tips of the treated albino Rats. The
treatments for 14 days. Liver enzyme assayed to know if the
extract had toxic effect on the liver cells
Determination of Blood glucose level were Alanine aminotransferase, (ALT),
All blood samples were collected by Aspatate aminetransferase (AST) and
cutting the tail-tips of the overnight fasted Alkaline phosphatase (ALT). The assays
rats. Blood samples for blood glucose were carried out using ultra violet (UV)
estimation were collected at intervals 0, kinetic kits produced by Cypress
2,4,8 and 24hrs following treatment. diagnostics. The test is based on
These tests were carried out using a photometric determination of rate of
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nicotinamide adenine' dinucleotide aminotransferase (AST) and Alkaline

(NADH) consuption by pyruvate and Phosphate (ALP) were 3-15IU/L, 5-18I u/L
oxaloacetate which is directly related to and 25-92IU/L respectively.
ALT and AST activities. At alkaline PH STATISTICAL ANALYSIS
alkaline phosphatase catalyses the Statistical analysis was carried out using
hydrolysis of p-nitrophenyl phosphate to statistical package for social sciences
yellow coloured p-nitro- phenolate and (SPSS) version 19. One way analysis of
phosphate; the change in absorbance variance (ANOVA) was adopted for
measured at 415nm is directly proportional comparison and the results were subjected
to the enzyme activity (table 4.3). The to post hoc test using least square
normal ranges for Alanine deviation (LSD). The data were expressed
aminotransferase (ALT), Aspartate as ± standard error of mean (SEM).
RESULTS

Determination of liver enzymes activities (IU/L)

Class ± ALT (IU/L) AST (IU/L) ALP (IU/L)
Normal Control 13.5±4.0 15.0±1.1 20.2±2.0
Diabetic control 16.0±2,0 20.5±1.0 100.2±.2.0
Diabetic+250mgGL 10.5±3.0 11.4±2.0 40.6±4.0
Diabetic+5mgGL 9.0±1.0 10.0±1.0 38.1±1.0
Diabetic+500mgGL 8.4±10 7.0±3.0 34.10±1.0
Diabetic+l000mgGL 6.2±10 5.1±1.0 30.20±1.0
ALT= Alanine amino transferase, AST = Aspartate aminotransferase, ALP=
Alkaline Phosphatase
Assay for liver enzymes namely ALT,AST and ALP is important in assessing optimal liver function in diabetics
treated with the extracts of Gongronema latifolium. The increase in ALT, AST and ALP were not statistically
different from the normal control group, indicating possible hepato-protective effect of the plant extracts. The
group that was administered the extract of Gongronema latifolium showed a significant reduction in ALT, AST
and ALP levels compared to the diabetic control.

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DISCUSSION which might contribute to its use as a

herbal therapy in the management of
The ASP, ALT and ALP levels support the diabetes.
fact that the extract played a protective
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