The Effect of Moringa oleifera Leaves Powder on Some Hormones to Prevent the Development of Experimental Hypothyroidism in Rabbits

1040 Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3

 The Effect of Moringa oleifera Leaves Powder on Some
 Hormones to Prevent the Development of Experimental
 Hypothyroidism in Rabbits

 Abeer Ataallah Ayyed Al –Hadidy1, Suher Taha Dawood2
 1
 Tutor Doctor, Department of Biology, College of Science, University of Mosul, Iraq, 2Research Scholar,
 Department of Biology, College of Science, University of Mosul, Iraq

 Abstract
 Background: Adipose tissue produces a many of active biological compounds, called adipokines that have
 role as an autocrine, paracrine, and endocrine method. The present study was carried out to investigate the
 effects of oral administration of Moringa oleifera leaves powder by which contains of active compounds
 to prevent the effect of carbimazole drug which used to induce and develop experimental hypothyroidism,
 in addition to understand the relationship between Visfatin hormone and other related hormones such as
 Resistn and thyroid hormones with or without using Moringa. Methods: Forty healthy male rabbits had
 been randomly distributed in to four groups, group1 had been orally drenched normal saline considered as
 a control group, group2, dealed with 300 mg Moringa powder /kg diet/day, group3 dealed with carbimazole
 (30 mg/ kg body weight/day) to initiate experimental hypothyroidism, group4 treated with 300 mg Moringa
 powder /kg diet/day with the carbimazole (30 mg/ kg body weight/day). Result: The results showed positive
 effects as a significant decrease (P≤ 0.01) in Visfatin, Resistin and Tumor necrosis Factor-α (TNF-α), with
 increase in level of T4 and Total Antioxidant Capacity (TAC) in rabbit’s that dealed with Moringa leaves
 powder. In contrast showed a negative effect as a significant increase in Visfatin, Resistin and TNF-α with
 decrease in level of T4 and TAC in rabbit’s that dealed with carbimazole, while the results tend in rabbits that
 given carbimazole with Moringa near with normal concentration in group1.

 Key words: Visfatin hormone, Resistin hormone, TNF-α.

 Introduction of adipokines is a consequence of abnormal plasma
 hormone levels and contributes some diseases
 Two decades ago, the first adipokine was discovered-
 development including atherosclerosis and diabetes
Leptin1 and adipose tissue began to be viewed also as
 mellitus II 3,4.
an endocrine organ2. Adipose cell produces a multiple
of active biological compounds, called dipocytokines Visfatin -an abbreviation of “visceral fat”-
such as (Adiponectin, Leptin, Resistin, and Visfatin), was described in 2004 5. This adipokine is mainly
that act in paracrine, an endocrine and an autocrine, produced in visceral fat tissues and only minimally
and manner. They have an important functions in in subcutaneous fat. Its production increases with
thermogenesis, reproductive functions, appetite control, increasing obesity, or rather with increasing visceral
and thyroid functions. These hormones have an effective fat. Secretion increases during hypoxia, inflammation,
function to control the metabolism of fats. Imbalance and hyperglycemia. It contributes to lipogenesis and
 the differentiation of adipocytes in visceral adipose
Corresponding Author: tissue, increasing their capacity to store fat6. Visfatin,
Dr. Abeer Ataallah Al-Hadidy also known as pre- B-cell colony-enhancing factor
Tutor Doctor, Department of Biology, as well as, nicotinamide phosphoribosyltransferase
College of Science, University of Mosul, (NAMPT) is a multifunctional protein with
E-mail ID: abesbio53@uomosul.edu.iq Contact No: suggested, immunological, metabolic and enzymatic
+9647718037256

Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3 1041

activities or properties. Visfatin has been analyzed Carbimazole was obtained from local pharmacies;
 in hyperthyroidism and hypothyroidism in vivo and in rabbits were treated by oral administration at dose 30
vitro studies, but findings are uncertain 7. mg/kg body weight/ day.

 In addition to Visfatin function in obesity, its also Standard ration
has other important biological functions. Visfatin has
 The basal diet for rabbits was prepared according
also been reported to be a cytokine that incites B-cell
 to National Research Council17, was prepared by Al-
development and maturation also delays neutrophil
 Ameen Company for poultry feed trade in Mosul city
apoptosis8 .
 with free water.
 Resistin hormone was discovered in mice in
 Rabbits
(2001), it means-resistance to insulin-, acquired
its name for its ability to resist or interfere with The study included (40) healthy male New Zealand
the role of insulin hormone9; at that time, it rabbits, ranging in age from 9-12 months and weighing
 was considered as a link between obesity and diabetes10. between 1250-1500 grams. These rabbits were obtained
In animal models, this link has been demonstrated from the College of Veterinary Medicine / University
conclusively, but in humans, the question of Resistin of Mosul and to ensure their pure strain. These rabbits
function seems to be more complicated. 11 are placed in custom cages (35 × 35 × 60 cm) with
 appropriate dimensions, provided with adequate
 Herbal medicine is nevertheless the mainstay in the
 ventilation under natural light- dark photoperiod. Rabbits
developing countries for principal health care. This is
 were fed a concentrated diet for a week for the purpose
specifically because of the universal trust that natural
 of adaptation, then after adaptation, it was divided into
tablets are beside any aspect of effects12. M. oleifera
 study groups.
is one of the world’s most useful plants. Every part of
the Moringa tree can be used for food and medicinal Moringa oleifera
purpose and are used for the treatment of rheumatism,
ascites, venomous bites and for improvement cardiac Packaged Moringa oleifera dry leaves powder
function13. Moringa tree was used by the ancient used for this study was purchased from the Moringa
Romans, Egyptians and Greeks, all parts are healthy plantation unit. National Research Center, Dokki, Egypt.
to eat and have long been consumed by humans, and Moringa powder was transported to Iraq for the purpose
their content of antioxidants give the plant’s image as a of research.
source of healthy food.14 It is usually named as Sainjana
 The study protocol
in India and Drumstick or Horseradish tree in English.15
 Forty healthy male rabbits were used in this study
 Different components of this plant comprise
 and were randomly assigned into four groups:
a profile of necessary minerals and are an
excellent source of protein, vitamins, amino acids Group 1 (Control group): included 10 male rabbits
 also rich sources of flavonols such as quercetin and fed an ordinary diet without any supplementation for an
kaempferol.16 A flavone, glycoflavone and acacetin, experimental period of 60 days. They were dosed with
Vitexin was also determined. The phenolic acids physiological saline to neutralize stress of catching
identified included p-coumaric acid, vanillic acid rabbits.
and melilotic acid.13 Therefore, this study has been
undertaken to investigate Moringa olifera effects Group 2: included 10 male rabbits fed an ordinary
against induced hypothyroidism, and estimation some diet mixed with 300 mg Moringa oleifera powder /kg
adipokines levels with or without treatment with this diet/day.18
plant. Group 3: included 10 male rabbits dealed with
 Material and Methods Carbimazole (30 mg/ kg body weight/day)19 to initiate
 experimental hypothyroidism and fed an ordinary diet
 Carbimazole (Anti-Thyroid drug)

1042 Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3

 Group 4: included 10 rabbits fed an ordinary diet alpha (TNF-α), interleukin-6 (IL-6), and C-reactive
mixed with 300 mg Moringa oleifera powder /kg diet/ protein (CRP). Depression is well known to occur in
day and (30 mg/ kg body weight/day) for 60 days. hypothyroidism. Depression is also characterized by
 increased inflammatory cytokines. This information
 Kits for Analysis
 interpret our results about significant increase of
 Commercial diagnostic kits for estimating Visfatin hormone level and TNF-α in group3.23 In
serum Visfatin and Resistin hormone and TAC were addition Visfatin released by visceral adipose tissue and
obtained from Elisa MyBioSource Company, USA. elevated in obesity 5,24, and obesity is an important sign
The serum TSH concentration was estimated using in hypothyroidism patients. 25
the Accubind ELISA Microwells Prepared Analysis
 Ozkaya et al. (2009) 26 tested the Visfatin values in
Kit, provided by Monobind Inc. American, based on
 Graves’ patients with hyperthyroidism andHashimoto’s
Immunoenzymometric assay. The concentration of
 thyroiditis patients with hypothyroidism and euthyroid
T4 and T3 hormones in serum was determined using
 healthy subjects before and after treatment. Hyperthyroid
the Accubind ELISA Microwells prepared assay kit,
 patients had significantly decreased Visfatin values
provided by Monobind Inc. by competitive enzyme
 compared with the hypothyroid group and control.
immunoassay.
 Moringa olifera extract had decreased the activity of
 Statistical Analysis IL-1, and TNF-α , and increased IL-2,27 by its component
 One-way ANOVA analysis of variance of many types of antioxidants,28 so this interpret the
using the General Linear Model, (SAS software) significant decrease of Visvatin level and TNF- α in
was used in the statistical analysis.20 To group 2 by depending upon the positive relationship
separate the means when significant differences between them.
exist Duncan used. Means and standard deviations were
 Some previous studies mentioned that elevated
calculated for all parameter. Difference were considered
 Visfatin value that have a role in gradual elevate of
significant at P ≤ 0.01.
 visceral fat and total abdominal fat and lead to elevate
 in pro-inflammatory factors TNF- α and IL-6 gene
 Results
 expression in fat cells 29,30, so this interpret our study
 The obtained results in Table (1) showed significant about increase the level of this parameters positively.
increase in serum Visfatin, Resistin hormone and TNF-α
in group3, in contrast decrease significantly in group2, In this study, Visfatin concentration depends on
while in group4 near the values of control group, T4 T3 and strongly on T4. This agreement with study that
hormone and TAC decrease significantly in group3, in explain Visfatin level in hypothyroidism depends on
contrast increase significantly in group2, while in group4 thyroid hormones value and might assume that factor
near the values of control group. should be taken into consideration to assess Visfatin
 level in patients with thyroid dysfunction.31 Ozkaya et al.
 Discussion (2009) 26 noted the significant negative relation between
 thyroid hormones and Visfatin. In vitro experiment
 Visfatin incites the production and
 showed the nonlinear regulation of Visfatin mRNA
secretion of pro-inflammatory cytokines (IL-
 expression in the T3-L1 cell culture model affected by
6, TNF- , and IL-1) and potentially play a role
 T3.32
as a chemotactic member for monocytes. In
addition, its expression is upregulated by Serum Visfatin values were negatively relation
IL-6, TNF- , and IL-1, promoted mRNA expression of with T4 values before treatment with Moringa olifera,
Visfatin. 21, 22 also there was no significant correlation between serum
 values of serum levels of TSH and T3 and Visfatin, this
 Hypothyroidism has been believed to be as an
 agreement with 33.
inflammatory disease characterized by increased
values of cytokines such as, tumor necrosis factor-

Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3 1043

 In addition, there is relationship between Resistin that the response was dose effective and the Moringa
hormone concentration and many inflammatory factors leaves extracts can be utilized in hypothyroidism status
including IL-6, TNF-α and C-reactive protein,34,35 this to normalize hormone levels, this agreement with
markers increased also with hypothyroidism. 23 This our results. The results is agreement with Ghadhban
statement supports the results of the current study. et al. (2019)43 who studies that Moringa extracts is
 necessary in hypothyroidism as it is elevating the T3
 Metwally et al (2017) 36 showed that Moringa oleifera
 and T4 hormone values and dropping TSH as well as
extract down-regulated mRNA expression of Resistin
 in hypercholesterolemia where it is reducing the total
and Leptin while it up-regulated Adiponectin gene
 cholesterol.
expression in fat rats relative to untreated obese control
counterparts. M. oleifera are mechanistically realized by The action of Carbimazole is to be as substrate
working directly on the adipokines of the visceral fat.36 for thyroid peroxidase (TPO) enzyme and inhibits it
This is what we have reached in this research regarding and reduction incorporation of iodide into tyrosine
the relationship between Moringa leaves and the Resistin molecules. In addition inhibits coupling of di-iodinated
hormone. and mono-iodinated to form thyroid hormones,44 this
 action interpret our result about significant decrease in
 In a study showed that Resistin value regulated via
 the level of T4.
T3 and T4, it was showed that Resistin value in patients
with hypothyroidism elevated and conversely, in The result in this study about TAC is harmony
patients with hyperthyroidism Resistin levels reduced. and compatible with the findings showed by Luqman
36. Chen et al. (2016) 37 mentioned there is a negatively
 et al. (2012)45 and Tuorkey (2016) 46 who showed that
relationship between Resistin and thyroid hormones in TAC elevated with an increase in Moringa oleifera
different thyroid states. This is what we reached in our concentration. It can be used to impulse the TAC of
current study. Some studies were observed that Resistin rabbits that are in the growth stage, so, Moringa oleifera
levels are variable in thyroid dysfunctions.38 may indicate to as a source of compounds with health
Higher Resistin levels in both hypothyroidism and protective with antioxidant activity. In this regard,
hyperthyroidism were observed compared to controls. Siddhuraju and Becker (2003)47 showed that Moringa
 leaves act as prebiotic effects and potentially antioxidant
 obesity is a sign of hypothyroidism status25 and
 phyto-chemicals, such as chlorogenic and caffeic acid.47
obesity is known to increase the release of several
 In addition, many studies which noticed that M. oleifera
cytokines and other cellular mediators, such as
 leaves are rich in polyphenols and flavonoids and have
interleukin IL-6 and IL-1 and TNF-α,.39 It is possible that
 very important antioxidant. 48,49
obesity induces Resistin expression. Our data recorded
significant decrease in the serum level of Resistin in the Mbikay (2012) 50 study agreement with our study
group4 treated with extract of Moringa oleifera. who said the Moringa oleifera supplementation into diet for
 the growing rabbits have low MDA concentration and high
 Anthocyanin compounds which present in Moringa
 TAC level, which may be due to a decreased reduction of
oleifera in adequate amount 40, accelerate Adiponectin
 the fat deposition of adipose by reduction the malate
production which increase thyroid hormones synthesis;
 dehydrogenase activity and lipoprotein lipase activity or
specially T4 hormone as a result of C- terminal globular
 elevating the hormone-sensitive lipase activity in the fat
reaction with receptors located in mitochondria of
 cells. These results may be attributable to the occurrence
thyroid gland cells.41
 of flavonoids compounds, which reduce oxidative
 Tabassum et al. (2013) 42 noticed in his study that the stress. The antioxidant capacity of Moringa leaves
group which given a large dose (500 mg/kg bw, 14days) being excellent and rich source of vitamin B, calcium,
of Moringa noticed maximum percentage elevate protein and potassium. The results are consistent
in hormone level of T3 and T4, where as a maximum with similar findings by Oseni and Idowu (2014)51
percentage reduction in TSH value was noticed when they reported that Moringa leaves increased antioxidant
compared to the other dose levels, which clearly proves enzymes like GSH, GSH-Px, catalase and TAC but,

1044 Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3

reduced MDA concentration in rats.51 Additionally, the Conclusion
antioxidant properties of Moringa leaves can be due
 From our study, we can conclude that M. oleifera
to the existence of glycosides, tannins, anthocyanin,
 is a good natural drug for it’s an effective role in
polyphenols and thiocarbamates, which activate
 improving experimentally induced hypothyrodism in
antioxidant enzymes, inhibit oxidases and eliminate
 rabbits, it is fighting against increasing Visfatin, Resistin
free radicals,45 so TNF-α decrease in group 2. Moringa
 and TNF-α by increasing the related hormone T4 and T3
leaves appears to contain a package of natural
 with increasing TAC which have important role against
antioxidant compounds like vitamin E, C, carotenoids
 TNF-α.
and polyphenols, which deserves further evaluation as
potential antioxidant agents,52 this interpret closing our Acknowledgment
results of TAC to normal level in group4.
 The authors express their grateful to the university
 Conflict of Interest: None of Mosul/ College of Science for their provided facilities,
 which assisted to improve the quality of this work.
 Source of Support: Self

 Table (1): The Hormonal values with TAC and TNF- values in groups under study

 Groups
 Group1 Group2 Group3 Group4
 Parameters

 Visfatin
 8.00±4.36b 3.88±2.7d 10.95±7.45a 6.70±4.41c
 ng/ml

 Resistin
 90.6± 3.6c 32.4± 5.4d 165.5± 2.7a 122.8± 6b
 Pg/ml

 TSH
 0.63±0.04a 0.60± 0.07a 0.67± 0.08a 0.66± 0.08a
 µIU/ml

 T4
 10.01±0.15b 13.72±2a 7.8±0.4d 8.8±0.7c
 µg/100ml

 T3
 1.1±0.04a 1.2±0.06a 0.96±0.04a 1.0±0.03a
 ng/ml

 TAC
 119±4.3b 139± 10.6a 85.1± 3.7d 100.5±8.4c
 ng/ml

 TNF-α
 16.1±1.4c 7.3±2.21d 85.2±1.5a 27.4±1.45b
 pg/ml

 The values is means ± SD, no. of rabbits/ group = 10

 The no. followed by different letters means there is significant difference.

 Ethical clearance: Taken from scientific committee of Biology department/ College of science/ Mosul University

Indian Journal of Forensic Medicine & Toxicology, July-September 2021, Vol. 15, No. 3 1045

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