South African Journal of Botany 129 (2020) 32–39

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South African Journal of Botany

journal homepage: www.elsevier.com/locate/sajb

Moringa oleifera Lam. prevents the development of high fructose

diet-induced fatty liver
N. Muhammad a,b,⁎, K.G. Ibrahim a,c, A.R. Ndhlala d, K.H. Erlwanger a
a
School of Physiology, Faculty of Health Sciences, University of the Witwatersrand, 7, York Road, Parktown, Johannesburg 2193, South Africa
b
Department of Physiology, College of Health Sciences, Federal University Birnin Kebbi, P.M.B. 1157 Birnin Kebbi, Nigeria
c
Department of Physiology, Faculty of Basic Medical Sciences, College of Health Sciences, Usmanu Danfodiyo University, P.M.B. 2254 Sokoto, Nigeria
d
Agricultural Research Council, Vegetable and Ornamental Plants (VOP), Private Bag X293, Pretoria 0001, South Africa

a r t i c l e i n f o a b s t r a c t

Article history: Ingestion of fructose-rich diets has been associated with the increase in the global prevalence of obesity, dyslipidemia
Received 11 July 2018 and non-alcoholic fatty liver disease across all age groups. Phytochemicals from the plant Moringa oleifera Lam. re-
Received in revised form 5 October 2018 portedly therapeutically reduce obesity, correct hyperlipidemia and decrease excess hepatic lipid accumulation in
Accepted 7 December 2018
adults. In light of the increasing prevalence of diet-induced metabolic dysfunction in children, this study was de-
Available online 8 January 2019
signed to investigate the protective effects of a methanolic leaf extract of M. oleifera on fructose-induced obesity
Edited by Prof Johannes van Staden and hepatic lipid accumulation in growing female rats. Fifty-one 21-day-old female Sprague Dawley rats were
weaned and randomly allocated to six treatment groups. The rats had ad libitum access to commercial standard
Keywords: rat food. In addition, they also had plain drinking water (negative control; C), or a 20% fructose solution (H), or
Fructose 400 mg.kg−1M. oleifera absolute methanolic leaf extract daily with or without fructose solution (M + H or Mo), or
Moringa oleifera 100 mg.kg−1 fenofibrate daily with or without fructose solution for drinking (F + H or FNF). Growth parameters, cir-
Fatty liver culating metabolites, visceral fat pads, hepatic lipid storage, and biochemical markers of general health were assessed
Triglycerides after 10 weeks of the interventions. A significant increase in circulating triglycerides (mmol.l−1) (2.30 ± 0.43 vs 1.70
Health
± 0.30; P b .01, ANOVA) and hepatic lipid stores (11.00 ± 0.21 vs 9.60 ± 0.21%; P b .05, ANOVA) was observed in
fructose-fed rats (H) compared with the negative (C) control. The methanolic leaf extracts of M. oleifera (M + H)
prevented the fructose induced elevation in hepatic lipid stores (9.80 ± 0.77 vs 11.00 ± 0.21%; P b .05, ANOVA). In
the fructose-fed rats the Moringa leaf extracts (M + H) and fenofibrate (F + H) did not prevent the fructose induced
hypertriglyceridemia (P b .05, ANOVA) observed in fructose only fed rats (H). However, rats supplemented with
M. oleifera leaf extracts alone (Mo) and fenofibrate alone (FNF) had similar triglyceride concentrations to the negative
control (C) (P N .05, ANOVA). Visceral fat pads (g) were significantly lower in the fenofibrate treated group (FNF)
compared to those on combined fructose and M. oleifera (M + H) (11.00 ± 3.00 vs 17.00 ± 3.40; P b .01, ANOVA).
Fructose intake did not result in a significant elevation (P N .05) in plasma cholesterol, however, rats on fenofibrate
alone had lower (P b .05) cholesterol concentrations compared to those on fructose alone and compared to the neg-
ative control group. Significantly elevated fasting blood glucose levels, tissue non-specific alkaline phosphatase activ-
ity, and hepatomegaly were observed only in fenofibrate treated groups (P ≤ .05, ANOVA). No significant differences
were observed in the serum insulin concentration (P N .05, ANOVA), HOMA-IR index (P N .05, ANOVA), plasma cre-
atinine (P N .05, ANOVA), body mass gain (P N .05, ANOVA), linear growth (tibia and femur length, P N .05), body mass
index (P N .05, ANOVA) and waist circumference (P N .05, ANOVA) between the different treatment groups. The pro-
phylactic potential of M. oleifera methanolic leaf extract should be considered as an alternative prophylactic natural
agent against non-alcoholic fatty liver disease, and thus needs to be further explored in humans.
© 2018 SAAB. Published by Elsevier B.V. All rights reserved.

1. Introduction cardiovascular disease, diabetes mellitus (Jayarathne et al., 2017) and

Central obesity and an abnormal lipid profile are risk factors for
metabolic syndrome and predispose to the development of
⁎ Corresponding author at: School of Physiology, Faculty of Health Sciences, University
of the Witwatersrand, 7, York Road, Parktown, Johannesburg 2193, South Africa.
E-mail: muhammad.nasiru@fubk.edu.ng (N. Muhammad).
non-alcoholic fatty liver disease (NAFLD) (Mamikutty et al., 2015). In
both developed and developing countries, the prevalence of obesity is
increasing rapidly and is not confined to adults, but also affects children
(Ahmed et al., 2014). The increase in the prevalence of obesity and
related lipid abnormalities has been associated with an increase in fruc-
tose consumption (Maarman et al., 2016) and lack of physical exercise
(Crescenzo et al., 2014) amongst other causes. The major dietary

https://doi.org/10.1016/j.sajb.2018.12.003
0254-6299/© 2018 SAAB. Published by Elsevier B.V. All rights reserved.
 N. Muhammad et al. / South African Journal of Botany 129 (2020) 32–39
sources of fructose are sucrose and High Fructose Corn Syrup (HFCS)
(Khitan and Kim, 2013) which are excessively consumed as sweeteners
in both developed and developing countries (Maarman et al., 2016).
Most of the ingested fructose is metabolized in the liver where it is
then stored as phospholipids, triacylglycerols and fatty acids
(Narayanan and Jesudoss, 2016). High-fructose diet induced
intrahepatic lipid accumulation leads to NAFLD (Mamikutty et al.,
2015), which progresses to non-alcoholic steatohepatitis (NASH) (De
Castro et al., 2013) and hepatocellular carcinoma (Sattar et al., 2014) if
uncontrolled. NAFLD is associated with increased adiposity, hyperlipid-
emia, insulin resistance and diabetes mellitus; and thus recognized as
the hepatic manifestation of metabolic syndrome (Narayanan and
Jesudoss, 2016). The NAFLD global prevalence is estimated at 76% in
the obese, 50% in diabetics while in the general population its preva-
lence is approximately 20% (Narayanan and Jesudoss, 2016).
Regular exercise and lifestyle modification are the first line approach
in the management of obesity and related lipid abnormalities (Ahmed
et al., 2014). This could be supplemented by drug therapy (Ahmed
et al., 2014) using fibrates, statins and niacin (Chapman et al., 2010)
amongst other interventions. However, these pharmacological agents
are associated with adverse side effects and potentially high costs
(Ahmed et al., 2014).
There is an increased interest in the use of herbal medicines and
phytochemicals which are considered to be a natural approach for en-
suring good health outcomes (Jain and Surana, 2015). Exploration of
plants for medicinal purposes has been encouraged by World Health Or-
ganization (WHO) (Jothi et al., 2016), and this has increased the interest
of scientific researchers in phytomedicines (Chowdhury et al., 2011).
The plant M. oleifera is widely exploited for its nutritional and medicinal
properties and benefits (Sangkitikomol et al., 2014).
Moringa oleifera Lam. (drumstick tree) belongs to a family called
Moringaceae of the angiosperm plants (Muhammad et al., 2016). It is
native to India but currently cultivated in Asia and other tropical coun-
tries (Muhammad et al., 2016). The leaves, flowers, immature pods,
seeds, bark and the roots of the plant have been widely used as food
and medicines in different disease conditions (Sangkitikomol et al.,
2014; Zvinorova et al., 2015). Animal studies using aqueous, alcohol or
hydroalcohol extracts of M. oleifera have revealed their hypolipidemic
and antiobesity properties (Stohs and Hartman, 2015). It was also re-
ported that methanolic leaf extracts of M. oleifera decrease serum cho-
lesterol and triacylglycerol concentrations in rats on a high fat diet
(Jain and Surana, 2015). Phytochemicals, macronutirents and
micronutrients present in M. oleifera leaves include β-carotene, protein,
calcium, potassium (Divi et al., 2012), starch, iron and vitamins A, B and
C (Zvinorova et al., 2015). The leaves of the plant are also rich in flavo-
noids, phenolics and carotenoids, all of which have biological activities
that are beneficial for metabolic health (Stohs and Hartman, 2015).
Most studies on the use of M. oleifera for obesity and diabetes man-
agement used adult male rats (Jain et al., 2010; Ahmed et al., 2014;
Bais et al., 2014). It is however notable that the risk factors for metabolic
syndrome including obesity and lipid abnormalities secondary to excess
fructose diets are not confined to adult males, they are also observed in
females and growing children (Maarman et al., 2016). There is thus need
to undertake studies which also target the lesser researched gender.
Given the high prevalence of dietary fructose-induced metabolic dys-
function in children, this study was aimed at investigating the protective
effects of a methanolic leaf extract of M. oleifera on high fructose diet-
induced obesity and hepatic lipid accumulation in growing female rats.
2. Materials and methods
2.1. Collection and processing of Moringa oleifera leaves
Fresh leaves of M. oleifera were harvested from the Agricultural
Research Council Research Farm, Roodeplaat, Pretoria, South Africa.
The plant was authenticated and a voucher specimen (Id:
33
Erlwanger 1) deposited at the Moss Herbarium of the University of
the Witwatersrand in Johannesburg, South Africa. Following desiccation
of the leaves in an oven (Salvis®, Salvis Lab, Schweiz, Switzerland) set at
40 °C, a blender (Waring Commercial Blender, HGB2WTG4, USA) was
used to comminute them into fine powder. The powder was extracted
with absolute methanol (Merck (pty) Ltd., South Africa) (100 g in
400 ml) on a shaker for 24 h. Previous studies have used a similar ratio
for alcohol solvent based extractions (Mahdi-Pour et al., 2012; Dutta,
2015). The mixture was then filtered through Whatmann No.1 filter
paper, after which the filtrate was concentrated using a rotary evapora-
tor (Buchi Rotavapor-R, Buchi Laboratoriums Technik AG, Schweiz,
Switzerland) at 48 °C. The condensed extract was then removed from
the rotary evaporator flask before it formed a thick paste and
oven-dried (Salvis®, Salvis Lab, Schweiz, Switzerland) at 40 °C for final
accelerated desiccation to prevent overgrowth by mold. The yield was
6%. The dried extracts were stored in dark tightly sealed bottles and
kept at 4 °C until use.
2.2. Animal experimental design
Ethical clearance for the study was obtained from the Animal Ethics
Screening Committee (AESC) of the University of the Witwatersrand
(Reference no: 2015/11/51/B). This research was conducted in accor-
dance with the internationally accepted principles for laboratory animal
use and care as found in South African National Standard (SANS
10386:2008 and Animals Protection Act, 1962: Act No. 71). Fifty one,
21-day-old weaned female Sprague Dawley rats were housed in a well
ventilated animal holding room in the Central Animal Services animal
research facility at the University of the Witwatersrand, Johannesburg,
South Africa. The rats were kept individually in standard rat polycarbon
cages containing clean wood shavings for bedding. The ambient
temperature in the room was maintained at 26 ± 2 °C. The lights in
the animal room were set on a 12 h lighting cycle (illumination was
turned on between 7 am–7 pm). The rats were allowed to acclimatize
to the environment for 4–6 days before commencing the intervention.
During the acclimatization period the rats were accustomed to being
weighed and fed standard commercially sourced rat chow, plain drink-
ing water and flavored gelatine cubes which would later be used as a ve-
hicle to deliver the treatments.
The rats weighing between 40 and 60 g were randomly divided into
six treatment groups. All groups received standard commercially sourced
rat chow (Epol, RSA) ad libitum for the whole duration of the 10-week
study. In addition, group I (control group; C; n = 10) received plain drink-
ing water and plain (flavored but no treatment) gelatine cubes. Group II
(H; n = 9) received 20% fructose solution as drinking fluid and plain gel-
atine cubes. Group III (M + H; n = 8) received 20% fructose solution as
drinking fluid and 400 mg.kg−1 body weight of methanolic extract of
M. oleifera (Bais et al., 2014). Group IV (F + H; n = 8) received 20% fruc-
tose solution to drink and 100 mg.kg−1 body weight of fenofibrate. Group
V (Mo; n = 8) received 400 mg.kg−1 body weight of methanolic extract
of M. oleifera and had plain drinking water. Group VI (FNF; n = 8) re-
ceived 100 mg.kg−1 body weight of fenofibrate and also had plain
water to drink. The M. oleifera extracts and fenofibrate (positive control
drug) were suspended in 2 ml flavored gelatine cubes. The 20% fructose
solution used in the study has previously been shown to cause obesity
and metabolic abnormalities in rats (Mamikutty et al., 2014) whilst the
dose of fenofibrate used (100 mg.kg−1) has also effectively been used
in rats to treat obesity (Ji et al., 2005). The gelatine cubes with or without
treatment were fed to the rats once a day. The drinking fluids (water or
20% fructose solution) were provided ad libitum.
2.3. Body mass measurement
The rats were weighed at the commencement of the study and then
twice weekly using a digital scale (Snowrex Electronic Scale, Clover
Scales, Johannesburg) to determine body mass gain (BMG) and monitor
34 N. Muhammad et al. / South African Journal of Botany 129 (2020) 32–39
growth performance in order to ensure that the rats were given the
treatments at the desired doses.
2.4. Terminal procedures
Following the 10-week treatment period, the rats were deprived of
food overnight for 12 h, but allowed access to plain drinking water.
The body masses of the rats were determined and two drops of blood
were taken following a puncture to the tail vein with a sterile 21G nee-
dle. The blood was used to ascertain the fasting glucose and triglyceride
concentrations. The fasting concentration of glucose in the blood was
quantified using a glucose meter (Ndong et al., 2007) (Contour Plus
Bayer Health Care, Diabetes Care, Isando, South Africa) according to
manufacturer's instructions. The triglyceride concentration was deter-
mined using a calibrated hand-held triglyceride meter (Accutrend,
Roche Diagnostics, Germany). The rats were then euthanased with an
intraperitoneal injection of sodium pentobarbitone (Centaur Laborato-
ries, Johannesburg, South Africa) at 150 mg.kg− 1 body weight. After
euthanasia, the waist (abdominal) circumference (WC) and body length
(nose–anus length) were measured using a measuring tape. The body
mass index (BMI) was subsequently computed according to the follow-
ing formula (Ahmed et al., 2014):
BMI(g. cm−2) = Body weight(g) ÷ Length2(cm2).
A ventral midline incision was made through the thorax and intra-
cardiac blood was drawn into syringes via attached needles. The blood
samples were then transferred into heparinized tubes (BD Vacutainer,
Plymouth, UK) and centrifuged (Rotofix 32A, Hettich Zentrifugen,
Germany) at 3700 rpm for 15 min. The supernatant (plasma) was
then transferred into micro-tubes (Greiner bio-one Diagnostics GmbH,
Austria) by pipette and kept at −20 °C for later assays. Following a mid-
line incision of the abdomen, the abdominal viscera were meticulously
removed. The contents of entire gastro-intestinal tract were carefully
emptied (Ndong et al., 2007). The masses of these tissues were
ascertained on an electronic balance (Presica 310M balance, Presica In-
struments AG, CH-Dietikon, Switzerland) (Zvinorova et al., 2015). The
lengths of small and large intestines were determined by stretching
them carefully on a cooled board and measured with a ruler
(Zvinorova et al., 2015). The liver was carefully removed from the ab-
dominal cavity and preserved at − 20 °C for later quantification of
lipid content. The abdominal visceral fat pads were gently removed
and their masses were ascertained on an electronic balance (Presica
310M balance, Presica Instruments AG, CH-Dietikon, Switzerland.
Empty carcasses following evisceration were also weighed (Snowrex
Electronic Scale, Clover Scales, Johannesburg).
2.5. Assessment of linear growth
The right femur with the attached tibia was dissected away from the
acetabulum, defleshed and disarticulated. An oven (Salvis®, Salvis Lab,
Schweiz, Switzerland) was used to dry the bones at 40 °C for 5 days
until the mass remained unchanged and then the weight established
on a Presica 310M balance (Zvinorova et al., 2015) to determine their
dry masses. The lengths of the bones were measured using KTV
150 mm Digital Caliper (MAJOR TECH (pty) LTD, Elandsfontein, South
Africa).
The bones' Seedor indexes were computed as described by Almeida
et al. (2008):
Seedor index = Bonemass(mg) ÷ Bonelength(mm).
2.6. Hepatic lipid content determination
A Tecator Soxtec apparatus for solvent extraction of lipids was used
according to standard methods (Official Methods of Analysis of Analyt-
ical Chemists, 2005). Briefly, the liver specimens were milled after
having been freeze-dried. A gram (1 g) was put into a pre-weighed
extraction thimble. The thimble was plugged with cotton wool (fat-
free) before being put on a thimble holder. Before being placed onto
the heating pads, petroleum ether was added to the extraction cups.
Four stages were employed for the extraction process: boiling for
30 min, rinsing for 30 min, recovery of the solvent for 10 min then fi-
nally desiccation for 30 min at 90 ± 5 °C.
2.7. Biochemical profile of general health
Plasma concentrations of cholesterol, blood urea nitrogen (BUN),
creatinine, tissue non-specific alkaline phosphatase (ALKP) and alanine
aminotransferase (ALT) were determined using a calibrated VetTest an-
alyzer (IDEXX VetTest® Clinical Chemistry Analyzer, IDEXX Laborato-
ries Inc., USA) as instructed by the manufacturer. The ALKP and ALT
were measured in units of activity.
Fasting serum insulin concentration (ng.ml− 1) was ascertained
using an Enzyme Linked Immuno-Sorbent Assay (ELISA) kit
(Elabscience Biotechnology Co., Ltd) (Crescenzo et al., 2014).
The insulin concentration (ng.ml− 1) was then expressed as
(μU.ml−1) using the formula: 1 ng.ml−1 is equivalent to 0.02 μU.ml−1.
The insulin resistance index was then computed according to the
Homeostasis Model of Assessment (HOMA-IR) using the following for-
mula (Divi et al., 2012):
HOMA − IR = fasting insulin (μU. ml − 1 ) × fasting glucose
(mmol. l− 1) ÷ 22.5.
2.8. Data analysis
GraphPad Prism 5.0v for windows (GraphPad Software, Inc. CA) was
used for data analysis. Data were expressed as mean ± standard devia-
tion. A one-way analysis of variance (ANOVA) was used to analyze the
data. A Bonferroni post hoc test was also done for comparison of the
means. Statistical significance was considered at P ≤ .05.
3. Results and discussion
3.1. Body mass, body mass gain, body mass index, waist circumference and
empty carcass mass
The initial and terminal body masses of the study rats are shown in
Fig. 1. The initial body masses of the rats were not significantly different
(P N .05) across all treatment groups. Following the 10 week interven-
tion, all the animals had a significant increase (P b .0001) in body
mass however, there was an insignificant difference observed (P N
.05) in the terminal body mass between the rats across the groups. No
***
400
C
H
300 M+H
Body mass (g)
F+H
200 Mo
FNF
100
0
Initial mass Terminal mass
Fig. 1. Initial and terminal body masses of the study rats *** = significantly different at
Pb0.0001; C = negative control (plain drinking water and plain gelatine cubes; n =
10); H = 20% fructose solution and plain gelatine cubes (n = 9); M + H = 400 mg.kg -
1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and 20% fructose
solution (n = 8); F + H = 100 mg.kg -1 of fenofibrate in gelatine cubes and 20%
fructose solution (n = 8); Mo = 400 mg.kg -1 of methanolic extract of Moringa oleifera
leaves in gelatine cubes and plain drinking water (n = 8); FNF = 100 mg.kg -1 of
fenofibrate in gelatine cubes and plain drinking water (n = 8).
 N. Muhammad et al. / South African Journal of Botany 129 (2020) 32–39 35

Table 1
Body mass gains, body mass indexes, waist circumferences and empty carcass masses of the study rats.

Parameter C H M+H F+H Mo FNF

BMG (%) 414.00 ± 64.00 403.00 ± 52.00 447.00 ± 55.00 424.00 ± 74.00 391.00 ± 53.00 402.00 ± 36.00
BMI (g.cm−2) 0.59 ± 0.03 0.58 ± 0.03 0.59 ± 0.03 0.60 ± 0.03 0.59 ± 0.05 0.56 ± 0.03
WC (cm) 16.00 ± 0.63 17.00 ± 0.94 17.00 ± 1.40 16.00 ± 0.62 17.00 ± 1.50 16.00 ± 0.52
Empty carcass (g) 225.00 ± 29.00 207.00 ± 11.00 219.00 ± 13.00 212.00 ± 12.00 212.00 ± 11.00 202.00 ± 12.00

P N .05; C = negative control (plain drinking water and plain gelatine cubes; n = 10); H = 20% fructose solution and plain gelatine cubes (n = 9); M + H = 400 mg.kg−1 of methanolic
extract of Moringa oleifera leaves in gelatine cubes and 20% fructose solution (n = 8); F + H = 100 mg.kg−1 of fenofibrate in gelatine cubes and 20% fructose solution (n = 8); Mo =
400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and plain drinking water (n = 8); FNF = 100 mg.kg−1 of fenofibrate in gelatine cubes and plain drinking
water (n = 8); BMG = body mass gain; BMI = body mass index; WC = waist circumference.

significant difference was observed (P N .05) in the BMG, BMI, WC and
empty carcass mass of the rats across different treatment groups
(Table 1) after the 10-week period of intervention.
High-fructose diet consumption over time may cause excess
body mass gain characterized by increased adiposity (Tappy
et al., 2010) which increases the risk of metabolic dysfunction. Ad-
iposity can be assessed by measuring the BMI and WC (Ahmed
et al., 2014; Crescenzo et al., 2014). In this 10-week study, fructose
consumption did not induce any significant effect on BMG, BMI
and WC of the rats (Table 1). This insignificant effect of fructose
on BMI and WC contrasted what Mamikutty et al. (2014) observed
following administration of similar 20% fructose solution on adult
Wistar rats for eight weeks. This difference may be due to the dif-
ferences in age and strain of rats used. In a different study,
M. oleifera caused a reduction in BMI and WC in rats (Ahmed
et al., 2014) on a high fat diet. In the current study, no significant
effect was induced by fenofibrate on BMG, BMI and WC of the rats
across all treatment groups (Table 1). It has been reported that FNF
reduces obesity and weight gain through binding to peroxisome
proliferator-activated receptor alpha (PPARα) thereby regulating
lipid metabolism (Ji et al., 2005).
The overall body mass of an animal can be affected by food in the
GIT, and hydration status, thus body mass may not be a good indicator
of long-term growth performance (Stookey, 2016). Although the rats
were fasted, it was notable that they still had residual content in the
GITs. Empty carcass mass thus provides a better indicator of growth per-
formance than intact body mass since the effect of GIT filling is avoided
(Owens et al., 1995). The current study observed no significant effect of
fructose, M. oleifera or fenofibrate on the empty carcass masses of the
study rats.
3.2. Long bones morphometry
Table 2 shows the masses, lengths and Seedor indexes of the femurs
and tibias of the rats across all the groups. The masses, lengths and
Seedor indexes of the rats' tibiae and femora were not significantly dif-
ferent (P˃0.05) across the treatment groups.
Linear growth, especially the tibial length, is a better indicator of
long-term growth performance than intact and empty carcass mass
(Zvinorova et al., 2015). The length of the bones during the growing
phase is not subject to acute changes, which can occur to body mass.
There is also a dose-dependent effect of growth hormone on long
bones (Eshet et al., 2004). It has been reported that long bones (femur
and tibia) morphometry is not adversely affected by fructose adminis-
tration even though some studies suggested that sugar intake has a
negative effect on bone mineralization in growing animals (Tsanzi
et al., 2008). In this study, the lengths, masses and Seedor indexes of
the femora and tibiae were not significantly affected by the different
treatments (Table 2).
3.3. Metabolites in circulation, insulin concentration and HOMA-IR index
Table 3 shows the concentrations of glucose, insulin, triglycerides
and cholesterol and the HOMA-IR indexes of the study rats. The rats
on fenofibrate and plain drinking water (FNF group) had significantly
higher (P b .0001) fasting blood glucose (FBG) versus those on
M. oleifera and fructose solution (M + H) and the control (C) groups
(P b .05). Those on M. oleifera and fructose (M + H) had significantly
lower (P b .05) FBG compared to those on fenofibrate and fructose
(F + H). The insulin concentrations in serum and computed HOMA-IR
indexes across all treatment groups were not significantly different
(P N .05). Fructose intake significantly increased (P b .05) the plasma
triglyceride concentration such that rats on fructose alone (H) had sig-
nificantly higher triglycerides concentration compared to the negative
controls (C) on normal feed (P b .01), M. oleifera alone (Mo, P b .0001)
and fenofibrate alone (FNF, P b .0001). Moringa oleifera leaf extracts
and fenofibrate did not prevent the fructose-induced hypertriglyc-
eridemia in the fructose-fed rats. Although fructose intake did not result
in a significant elevation (P N .05) in plasma cholesterol, rats on
fenofibrate alone (FNF) had lower (P b .05) cholesterol concentrations
compared to those on fructose alone (H) and those which served as
the negative control group (C).
Plasma glucose metabolism can be increased by excess fructose
consumption (Lê and Tappy, 2006). However, in the current study we
observed that no significant effect was exerted by either fructose or
M. oleifera on fasting blood glucose (FBG) across the different treat-
ments (Table 3).
The metabolism of carbohydrates and lipids is regulated by insulin, a
hormone that maintains normoglycemia by enhancing glucose uptake
in peripheral tissues (Wilcox, 2005). The HOMA-IR index is used to as-
sess insulin sensitivity; the higher the index the lower the insulin sensi-
tivity (Divi et al., 2012). High dietary fructose intake is linked with

Table 2
The masses, lengths and Seedor indexes of rats' femora and tibiae.

Parameter C H M+H F+H Mo FNF

Tibial mass (mg) 376.00 ± 19.00 364.00 ± 22.00 364.00 ± 11.00 372.00 ± 27.00 368.00 ± 15.00 352.00 ± 15.00
Tibial length (mm) 37.00 ± 0.26 36.00 ± 0.42 37.00 ± 0.78 36.00 ± 0.50a 36.00 ± 0.55 36.00 ± 0.58
Tibial Seedor index 10.00 ± 0.49 10.00 ± 0.53 9.90 ± 0.22 10.00 ± 0.62 10.00 ± 0.46 9.70 ± 0.29
Femoral mass (mg) 548.00 ± 42.00 521.00 ± 23.00 528.00 ± 36.00 548.00 ± 45.00 542.00 ± 27.00 517.00 ± 14.00
Femoral length (mm) 33.00 ± 0.98 32.00 ± 0.79 33.00 ± 1.50 33.00 ± 0.96 33.00 ± 0.89 32.00 ± 0.72
Femoral Seedor index 16.00 ± 0.87 16.00 ± 0.79 16.00 ± 0.56 17.00 ± 1.00 16.00 ± 0.80 16.00 ± 0.51

P N .05; C = negative control (plain drinking water and plain gelatine cubes; n = 10); H = 20% fructose solution and plain gelatine cubes (n = 9); M + H = 400 mg.kg−1 of
methanolic extract of Moringa oleifera leaves in gelatine cubes and 20% fructose solution (n = 8); F + H = 100 mg.kg−1 of fenofibrate in gelatine cubes and 20% fructose solution
(n = 8); Mo = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and plain drinking water (n = 8); FNF = 100 mg.kg−1 of fenofibrate in gelatine cubes
and plain drinking water (n = 8).
36 N. Muhammad et al. / South African Journal of Botany 129 (2020) 32–39
Table 3
Concentrations of blood glucose, insulin, triglycerides and cholesterol, and HOMA-IR index of the study rats.
Parameter C H
Glucose (mmol.l−1) 4.10 ± 0.26ab 4.20 ± 0.38ab
Insulin (ng.ml−1) 23.00 ± 9.90a 23.00 ± 19.00a
HOMA-IR Index 4.20 ± 1.60a 4.40 ± 3.60a
Triglycerides (mmol.l−1) 1.70 ± 0.30a 2.30 ± 0.43b
Cholesterol (mg.dl−1) 77.00 ± 10.00a 78.00 ± 12.00a
abc = within row means with different letters in superscripts significantly different at P ≤ .05; C = negative control (plain drinking water and plain gelatine cubes; n = 10); H = 20%
fructose solution and plain gelatine cubes (n = 9); M + H = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and 20% fructose solution (n = 8); F + H
= 100 mg.kg−1 of fenofibrate in gelatine cubes and 20% fructose solution (n = 8); Mo = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and plain drinking
water (n = 8); FNF = 100 mg.kg−1 of fenofibrate in gelatine cubes and plain drinking water (n = 8); HOMA-IR = Homeostatic model of insulin resistance.
elevations in plasma C-peptide levels that affect insulin sensitivity
thereby leading to insulin resistance (Lê and Tappy, 2006). The dietary
fructose also interferes with insulin receptor tyrosine phosphorylation
in the brain leading to insulin resistance (Lê and Tappy, 2006). However,
in the current study, insignificant differences in the insulin concentra-
tions and the HOMA-IR indexes were observed across all treatment
groups (Table 3). This showed that drinking a 20% fructose solution
did not result in insulin-mediated metabolic dysfunction. This could
be due to the age of the rats as they may have channeled most of the ex-
cess energy from fructose into growth.
Both short- and long-term consumption of excess dietary fructose in
rodents can lead to elevated plasma triglycerides (TGs) concentrations
in rodents (Lê and Tappy, 2006). Higher TGs concentrations were in-
deed recorded in our fructose treated groups compared to negative
controls (Table 3) in this study. Moringa oleifera leaf extracts and
fenofibrate did not prevent the fructose induced hypertriglyceridemia
in the fructose fed rats. The rats on M. oleifera alone (Mo) and
fenofibrate alone (FNF) had similar concentrations of circulating tri-
glycerides with the negative controls (C). It has previously been re-
ported that in rats on a high fat diet, M. oleifera decreased TGs via its
phytochemicals (saponins and tannins) that inhibit fat utilization
(Ahmed et al., 2014). Fenofibrate binds to, and activate its agonist, per-
oxisome proliferator-activated receptor alpha (PPARα) that regulate
lipid and lipoprotein metabolism (Jeong et al., 2004; Ji et al., 2005).
This leads to intracellular lipid breakdown (Jeong and Yoon, 2009).
The consumption of fructose-rich diets leads to the formation of very
low density lipoprotein cholesterol (VLDL-c) (Elliott et al., 2002). How-
ever, fructose did not significantly elevate plasma cholesterol in this
study, though we did not measure the cholesterol subtypes (Table 3).
It was also observed in this study that rats on fenofibrate alone had
lower cholesterol concentrations compared to those on fructose alone
and the negative control group. This could be supported by the fact
that fenofibrate decreases cholesterol absorption in the gut and in-
creases cholesterol excretion in feces (Valasek et al., 2007).
3.4. Visceral fat pad masses
The masses of the rats' abdominal visceral fat pads following the
10 week treatment period are shown in Table 4. Rats that received
fenofibrate alone had a significantly lower (P b .01) absolute and rela-
tive (to tibial length) visceral fat mass compared to those on fructose
and treated with M. oleifera (M + H). There were no significant differ-
ences (P N .005) observed between the rats in the other groups.
It has been shown that fructose-rich diets enhance visceral fat depo-
sition, and this predisposes to the development of metabolic syndrome
(Crescenzo et al., 2014). The current study observed that no significant
effect was induced by either fructose or M. oleifera (Table 4). This
could be due to the age of rats as stated earlier. Previous studies on
the development of dietary-induced obesity have shown its develop-
ment after 100 days (Patel and Srinivasan, 2010). However, rats that re-
ceived fenofibrate alone had a significantly lower (P b .01) absolute and
relative (to tibial length) visceral fat mass compared to those on
M+H F+H Mo FNF
3.90 ± 0.31b 4.40 ± 0.40ac 4.10 ± 0.34ab 4.60 ± 0.23ac
17.00 ± 9.50a 22.00 ± 12.00a 24.00 ± 20.00a 19.00 ± 12.00a
2.80 ± 1.60a 4.40 ± 2.50a 4.70 ± 4.30a 3.90 ± 2.40a
2.20 ± 0.38b 2.40 ± 0.25b 1.50 ± 0.22a 1.50 ± 0.16a
73.00 ± 10.00ab 75.00 ± 12.00ab 70.00 ± 9.50ab 60.00 ± 12.00b
combined fructose and M. oleifera. This is in line with the pharmacolog-
ical effects of fenofibrate on body fat stores (Jeong and Yoon, 2009).
3.5. Hepatic storage of lipids
Fig. 2 shows the hepatic lipid storage of the study rats following the
10-week period of intervention. Rats on fructose alone (H) had signifi-
cantly more (P b .05) liver lipid content in comparison to the negative
controls (C) and the rats on M. oleifera (Mo) alone (P b .05). Moringa
oleifera significantly reduced the fructose-induced accumulation of
hepatic lipids (M + H vs H; P b .05). Treatment with fenofibrate
significantly reduced fructose-induced lipid accumulation (F + H vs
H) to a significantly greater extent than M. oleifera (F + H vs M + H;
P b .01). Rats on fenofibrate alone had significantly lower liver lipid con-
tent compared to all other groups (P b .0001) except those on M. oleifera
alone. Rats on M. oleifera alone also had significantly lower lipid content
compared to all other groups (P b .05).
The results of the current study showed that 20% dietary fructose
solution induced a significant increase in hepatic lipids in comparison
to control groups of rats (Fig. 2). Hepatic lipid content reportedly
increases within 6 weeks of high-fructose diet consumption in rats
(Tappy et al., 2010). However, a major and important finding for this
study was that daily administration of methanolic extracts of
M. oleifera leaves (400 mg.kg−1 body weight) was able to prevent the
effect of fructose on hepatic lipid stores. The prophylactic effect of
M. oleifera might be attributed to β-sitosterol which indirectly decreases
cholesterol levels (Divi et al., 2012), as well as saponins and tannins that
inhibit dietary fat utilization (Ahmed et al., 2014). There may also be
other mechanisms of action of the Moringa extracts on liver lipid metab-
olism and these need to be explored.
The effect of fenofibrate was more pronounced than that of
M. oleifera. Fenofibrate was able to decrease hepatic lipid storage likely
due its lipid lowering effect via binding to its nuclear receptor and
inhibiting lipid metabolism (Ji et al., 2005).
3.6. Organs of the gastro-intestinal tract (GIT)
Table 5 shows the morphometry of the GIT organs of the study rats.
There were no significant differences in the absolute and relative
masses of GIT visceral organs (P N .05) across all treatment groups.
Phytotherapy can affect the growth performance of an animal via
altering the GIT morphometry (Pérez et al., 2007). In the current
study, no significant effect was induced by administration of fructose,
fenofibrate or M. oleifera on the GIT visceral organs except the cecum.
The cecum plays a major role in rats through fermentation of polysac-
charides to short chain fatty acids, and this generates more energy for
body requirements (Pascoal et al., 2013; Zvinorova et al., 2015). The
heavier cecal mass observed in M. oleifera treated groups in this study
could be due to the polysaccharides present in the methanolic extract
of the plant, which could have increased the number of cecal mucosal
cells following fermentation process (Dangarembizi et al., 2014). Simi-
lar effects on the cecum were also observed with plant extracts by
other investigators (Erlwanger and Cooper, 2008; Beya et al., 2012;
 N. Muhammad et al. / South African Journal of Botany 129 (2020) 32–39 37

Table 4
Abdominal visceral fat pad masses of the study rats.

Fat pad mass C H M+H F+H Mo FNF

Visceral (g) 14.00 ± 2.10ab 15.00 ± 2.60ab 17.00 ± 3.40a 13.00 ± 3.70ab 14.00 ± 2.00ab 11.00 ± 3.00b
Visceral (grTL) 0.38 ± 0.06ab 0.42 ± 0.07ab 0.46 ± 0.09a 0.34 ± 0.10ab 0.39 ± 0.06ab 0.31 ± 0.09b

ab = within row means with different letters in superscripts significantly different at P ≤ .05; C = negative control (plain drinking water and plain gelatine cubes; n = 10); H = 20% fruc-
tose solution and plain gelatine cubes (n = 9); M + H = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and 20% fructose solution (n = 8); F + H = 100 mg.
kg−1 of fenofibrate in gelatine cubes and 20% fructose solution (n = 8); Mo = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and plain drinking water (n =
8); FNF = 100 mg.kg−1 of fenofibrate in gelatine cubes and plain drinking water (n = 8); grTL = mass relative to tibial length.

Zvinorova et al., 2015). However the increased cecal mass did not affect
growth performance of the rats in the current study.
3.7. GIT accessory organs, heart and kidneys
Table 6 shows the masses of the liver, pancreas, heart and kidneys
of the study rats. Rats receiving fenofibrate with or without fructose,
had absolute and relative liver masses that were significantly heavier
(P b .0001) in comparison to other groups. Rats on combined fructose
and fenofibrate had significantly higher (P b .0001) absolute kidney
masses compared to control and those on fructose alone. Those on
combined fructose and fenofibrate had significantly higher (P b .05)
absolute kidney masses compared to those on M. oleifera alone.
Rats on combined fructose and fenofibrate had significantly higher
(P b .05) kidney masses relative to tibial length versus the control
group. Insignificant differences were observed in the absolute and rela-
tive masses of heart and pancreas across different treatments (P N .05).
High fructose diets have been shown to cause cardiovascular, renal
and hepatic complications that affect their individual organ masses
(Dachani et al., 2012). Although there is a belief that ‘natural is better
than synthetic’, phytotherapy is not free from unwanted side effects
(Oyagbemi et al., 2013). Hepatic and kidney damage was reported fol-
lowing chronic administration of methanolic leaf extract of M. oleifera
(400 mg.kg−1 body weight) to adult Wistar rats for eight weeks
(Oyagbemi et al., 2013). Fenofibrate treatment is associated with hepa-
tomegaly (Ji et al., 2005). Hence, it was important to assess functional
health status of the vital organs in this study. Administration of 20%
fructose solution in this study had no impact on the absolute and rela-
tive masses of the heart, pancreas, liver and kidney (Table 6). No signif-
icant effect was observed in the masses of these organs in M. oleifera
treated groups. However, the absolute and relative masses of the liver
***
15
* ALT is an enzyme found in the liver cells, and is a useful marker in
Lipid content (% liver mass)
and kidneys were higher in the fenofibrate groups. Previous studies
have reported that fenofibrate treatment is associated with hepatomeg-
aly in male Sprague Dawley rats (Ji et al., 2005; Hong et al., 2007), a find-
ing which has been attributed to the inflammatory effect of fenofibrate
on the liver (Tolman, 2000). The increase in kidney mass seen in the
fenofibrate treated groups could be attributed to adverse drug reaction
of fenofibrate on the kidney, since the drug has been shown to be neph-
rotoxic (Attridge et al., 2013).
3.8. General health biochemical markers
Table 7 shows the activity of the liver enzyme ALT, the tissue non-
specific ALKP; and the concentrations of biomarkers of kidney function:
creatinine, BUN and urea to creatinine ratio. Rats on combined fructose
and fenofibrate (F + H) had significantly higher ALKP activity compared
to negative control (P b .01), those on M. oleifera alone (Mo, P b .01)
those on combined fructose and M. oleifera (M + H, P b .01), and those
on fructose alone (H, P b .05). Those on fenofibrate alone (FNF) had sig-
nificantly greater (P b .05) ALKP activity compared to either those on
M. oleifera alone (Mo) or those on combined fructose and M. oleifera
(M + H). Rats on fructose and treated with fenofibrate (F + H) also
had significantly higher (P b .05) ALT activity compared to those on
M. oleifera alone (Mo). Compared to the negative controls (C) a signifi-
cant decrease in BUN concentrations compared to those on M. oleifera
alone (Mo, P b .0001), fructose alone (H, P b .01) and combined fructose
and FNF (F + H, P b .05) was recorded. A significantly lower (P b .01)
urea to creatinine ratio of the rats on either fructose alone (H, P b .01)
or M. oleifera alone (Mo, P b .05) in comparison to the negative controls
(C) was seen. The differences in the plasma creatinine concentrations
across different treatment groups were insignificant (P N .05).
Monitoring the health of animals undergoing interventional studies
is necessary so that the toxicity, safety and the risks associated with the
interventions can be assessed (Oyagbemi et al., 2013). In view of this,
the surrogate markers of function of vital metabolic organs, specifically
the liver and kidney were assessed in this study.

* liver damage conditions (Rajesh et al., 2009; Sharifudin et al., 2013).

**
The enzyme ALKP is found in the cells lining the liver biliary ducts
10 and it abnormally rises in conditions associated with bile duct dam-
age/obstruction. Unlike ALT, ALKP is not specific to the liver as its
levels are also elevated in conditions associated with osteoblast ac-
tivity and pregnancy (Schoch and Whiteman, 2007). Intestinal
5
cells of the GIT also produce ALKP (Thapa and Walia, 2007). A strong
relationship exists between the components of metabolic syndrome,
NAFLD and NASH (Kerner et al., 2005). It has been reported that
0 prolonged ingestion of high-fructose diets predisposes to NAFLD in
C H M+H F+H Mo FNF rats (De Castro et al., 2013). These liver diseases are associated
Treatment Groups with abnormal high levels of ALT and to a lesser extent, ALKP
(Kerner et al., 2005) which are the surrogate markers of liver func-
Fig. 2. Hepatic lipid storage of the study rats *** = Pb0.0001; ** = PPb0.01; * = PPb0.05. C tion. In this study, administration of 20% fructose solution did not af-
= negative control (plain drinking water and plain gelatine cubes; n = 10); H = 20%
fect the plasma activities of either ALT or ALKP of the study rats
fructose solution and plain gelatine cubes (n = 9); M + H = 400 mg.kg -1 of
methanolic extract of Moringa oleifera leaves in gelatine cubes and 20% fructose solution (Table 7). However, it was observed that rats on fenofibrate with
(n = 8); F + H = 100 mg.kg -1 of fenofibrate in gelatine cubes and 20% fructose or without fructose had higher activities of tissue non-specific
solution (n = 8); Mo = 400 mg.kg -1 of methanolic extract of Moringa oleifera leaves ALKP compared to other groups. As the ALKP measured was tissue
in gelatine cubes and plain drinking water (n = 8); FNF = 100 mg.kg -1 of fenofibrate non-specific, the elevation could have been due to the effect of
in gelatine cubes and plain drinking water (n = 8).
fenofibrate which increases osteoblast activity leading to elevated
38 N. Muhammad et al. / South African Journal of Botany 129 (2020) 32–39

Table 5
The masses and lengths of GIT segments.

Organ C H M+H F+H Mo FNF

Stomach (g) 1.50 ± 0.13 1.50 ± 0.15 1.50 ± 0.11 1.50 ± 0.23 1.50 ± 0.10 1.50 ± 0.10
Stomach (grTL) 0.04 ± 0.00 0.04 ± 0.01 0.04 ± 0.00 0.04 ± 0.01 0.04 ± 0.00 0.04 ± 0.00
S.I. (g) 6.80 ± 0.52 6.70 ± 0.47 7.10 ± 0.64 7.30 ± 0.64 6.80 ± 0.38 6.90 ± 0.37
S.I. (grTL) 0.18 ± 0.02 0.19 ± 0.01 0.19 ± 0.02 0.20 ± 0.02 0.19 ± 0.01 0.19 ± 0.01
S.I. (mm) 1247 ± 88.00 1229 ± 103.00 1299 ± 122.00 1233 ± 44.00 1215 ± 89.00 1259 ± 73.00
L.I. (g) 1.70 ± 0.14 1.50 ± 0.19 1.50 ± 0.25 1.70 ± 0.24 1.70 ± 0.15 1.60 ± 0.22
L.I. (grTL) 0.05 ± 0.01 0.04 ± 0.01 0.04 ± 0.01 0.05 ± 0.01 0.05 ± 0.00 0.05 ± 0.01
L.I. (mm) 221.00 ± 19.00 213.00 ± 6.70 214.00 ± 18.00 219.00 ± 8.30 225.00 ± 9.30 221.00 ± 21.00
Cecum (g) 1.10 ± 0.19 1.00 ± 0.16 0.97 ± 0.12 1.10 ± 0.26 1.10 ± 0.16 1.00 ± 0.14
Cecum (grTL) 0.03 ± 0.01 0.03 ± 0.01 0.03 ± 0.00 0.03 ± 0.01 0.03 ± 0.01 0.03 ± 0.01

P N .05; C = negative control (plain drinking water and plain gelatine cubes) (n = 10); H = 20% fructose solution and plain gelatine cubes (n = 9); M + H = 400 mg.kg−1 of methanolic
extract of Moringa oleifera leaves in gelatine cubes and 20% fructose solution (n = 8); F + H = 100 mg.kg−1 of fenofibrate in gelatine cubes and 20% fructose solution (n = 8);
Mo = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and plain drinking water (n = 8); FNF = 100 mg.kg−1 of fenofibrate in gelatine cubes and plain
drinking water (n = 8); grTL = mass relative to tibial length; S.I. = small intestines; L.I. = large intestines.

Table 6
The masses of liver, pancreas, heart and kidneys.

Organ C H M+H F+H Mo FNF

a a a a a
Heart (g) 1.00 ± 0.10 1.00 ± 0.05 1.00 ± 0.11 1.10 ± 0.13 1.10 ± 0.12 0.99 ± 0.04a
Heart (grTL) 0.03 ± 0.00a 0.03 ± 0.00a 0.03 ± 0.00a 0.03 ± 0.00a 0.03 ± 0.00a 0.03 ± 0.00a
Liver (g) 7.60 ± 0.57a 7.80 ± 0.71a 8.30 ± 0.58a 13.00 ± 1.20b 7.50 ± 0.67a 11.00 ± 1.30b
Liver (grTL) 0.21 ± 0.01a 0.22 ± 0.02a 0.23 ± 0.01a 0.36 ± 0.03b 0.21 ± 0.02a 0.30 ± 0.04b
Pancreas (g) 1.20 ± 0.23a 1.10 ± 0.23a 1.10 ± 0.12a 1.20 ± 0.16a 1.10 ± 0.20a 1.20 ± 0.14a
Pancreas (grTL) 0.03 ± 0.01a 0.03 ± 0.01a 0.03 ± 0.01a 0.03 ± 0.00a 0.03 ± 0.01a 0.03 ± 0.01a
Kidneys (g) 1.70 ± 0.14a 1.70 ± 0.12a 1.80 ± 0.11ab 2.00 ± 0.11b 1.70 ± 0.11a 1.80 ± 0.10ab
Kidneys (grTL) 0.05 ± 0.0a 0.05 ± 0.00ab 0.05 ± 0.00ab 0.05 ± 0.00b 0.05 ± 0.01ab 0.05 ± 0.00ab

ab = within row means with different superscript significantly different at P ≤ .05; C = negative control (plain drinking water and plain gelatine cubes; n = 10); H = 20% fructose solution
and plain gelatine cubes (n = 9); M + H = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and 20% fructose solution (n = 8); F + H = 100 mg.kg−1 of
fenofibrate in gelatine cubes and 20% fructose solution (n = 8); Mo = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and plain drinking water (n = 8); FNF
= 100 mg.kg−1 of fenofibrate in gelatine cubes and plain drinking water (n = 8); grTL = mass relative to tibial length.

Table 7
Surrogate markers of general health.

Parameter C H M+H F+H Mo FNF

ALT (U.l−1) 58.00 ± 3.20ab 53.00 ± 4.90ab 53.00 ± 1.90ab 60.00 ± 6.80a 51.00 ± 3.10b 58.00 ± 9.30ab
ALKP (U.l−1) 107.00 ± 33.00ac 109.00 ± 30.00ac 96.00 ± 15.00a 174.00 ± 55.00b 98.00 ± 12.00a 159.00 ± 49.00bc
Creatinine (mg.dl−1) 0.58 ± 0.05a 0.61 ± 0.06a 0.58 ± 0.07a 0.60 ± 0.11a 0.60 ± 0.11a 0.54 ± 0.09a
BUN (mg.dl−1) 19.00 ± 1.60a 16.00 ± 2.30b 17.00 ± 2.10ab 16.00 ± 2.00b 15.00 ± 1.20b 17.00 ± 2.20ab
Ur:Cr 34.00 ± 5.40a 25.00 ± 2.80b 29.00 ± 3.70ab 27.00 ± 5.20ab 26.00 ± 5.10b 32.00 ± 4.60ab

abc = within row means with different superscript significantly different at P ≤ .05; C = negative control (plain drinking water and plain gelatine cubes; n = 10); H = 20% fructose so-
lution and plain gelatine cubes (n = 9); M + H = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and 20% fructose solution (n = 8); F + H = 100 mg.kg−1
of fenofibrate in gelatine cubes and 20% fructose solution (n = 8); Mo = 400 mg.kg−1 of methanolic extract of Moringa oleifera leaves in gelatine cubes and plain drinking water (n = 8);
FNF = 100 mg.kg−1 of fenofibrate in gelatine cubes and plain drinking water (n = 8); ALT = alanine aminotransferase; ALKP = alkaline phosphatase; BUN = blood urea nitrogen; Ur:Cr
= urea to creatinine ratio.

plasma ALKP concentrations (Syversen et al., 2009). However, hepa-
tomegaly was also observed in fenofibrate treated groups thus it is
unclear what the source was of the ALKP as the assay used was un-
able to differentiate between the isoforms of ALKP.
Increased plasma concentrations of urea and creatinine as well as
urea to creatinine ratio have been considered as indicators of renal dam-
age (Oyagbemi et al., 2013). Previous studies have shown that chronic
fructose ingestion predisposes to renal dysfunction (Nakayama et al.,
2010). We found that neither administration of 20% fructose solution
nor that of M. oleifera leaf extract (400 mg.kg−1) for 10 weeks caused
any renal impairment as assessed by creatinine and urea in the study
rats.
4. Conclusion
Administration of 20% fructose solution to female Sprague Dawley
rats for 10 weeks elevated hepatic lipid content and triglycerides con-
centrations without affecting their growth performance. The elevation
in hepatic lipid content was prevented by oral administration of meth-
anolic extracts of M. oleifera leaves. This shows that the use of
phytomedicines like M. oleifera leaf extracts could be a better alternative
in managing fatty liver disease than synthetic agents like fenofibrate
since no adverse effects from the plant were observed in this study.
The prophylactic potential of M. oleifera methanolic leaf extracts against
hypertriglyceridemia and fatty liver in high-fructose fed observed in the
rats needs to be explored in humans.
Conflict of interest
There is no conflict of interest regarding the publication of the
manuscript.
Acknowledgements
We thankfully acknowledge the Medical Faculty Research Endow-
ment Fund, Faculty of Health Sciences Research Committee (Grant No:
001.401.8521101…PHSLMFR), and the National Research Foundation
of South Africa (Grant No: IFR 2010041900009); Federal University
Birnin Kebbi (Nigeria) and Tertiary Education Trust Fund of Nigeria for
making funds available for the research. The authors also wish to thank
 N. Muhammad et al. / South African Journal of Botany 129 (2020) 32–39
the staff of the Central Animal Service Unit, Faculty of Health Sciences,
University of the Witwatersrand for care and handling animals and De-
partment of Science and Technology, South African Government for
supporting Moringa research.
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