Received: 9 February 2021 | Revised: 29 April 2021 | Accepted: 7 May 2021

DOI: 10.1111/jfbc.13801

ORIGINAL ARTICLE

The positive effect of black seed (Nigella sativa L.) essential

oil on thyroid hormones in rats with hypothyroidism and
hyperthyroidism

Gulcan Avci1 | Elmas Ulutas2 | Vural Ozdemir3 | Ibrahim Kivrak4 | Aziz Bulbul5

1
Department of Biochemistry, Faculty of
Veterinary Medicine, University of Afyon Abstract
Kocatepe, Afyonkarahisar, Turkey In our study, the effect of essential oil obtained from Nigella sativa L. (NSE) on thyroid
2
Department of Physiology, Faculty of
hormones and antioxidant balance in hypothyroidism (HT) and hyperthyroidism
Veterinary Medicine, University of Bozok,
Yozgat, Turkey (HP) models induced by propylthiouracil(PTU) and L-­thyroxine(LT4), respectively, in
3
Department of Anatomy, Faculty of rats were investigated for 4 weeks. NSE was administered by gastric gavage at a
Veterinary Medicine, University of Afyon
Kocatepe, Afyonkarahisar, Turkey
dose of 200 mg/kg body weight. In this study, 48 male Wistar albino rats with an
4
Food Analysis Application, Research average weight of 180–­290 g and age 5–­6 months were divided into eight groups, as
Center, University of Mugla Sıtkı Kocman, follows: groups with HT, (1) control, (2) HT, (3) NSE, and (4) HT + NSE; groups with
Mugla, Turkey
5 HP, (1) control, (2) HP, (3), and NSE (4) HP + NSE. As a result, we found that NSE
Department of Physiology, Faculty of Milas
Veterinary Medicine, University of Mugla administration increased total triiodothyronine (TT3) and decreased nitric oxide in
Sıtkı Kocman, Mugla, Turkey
HT + NSE. Besides, it decreased TT3 in HP + NSE and increased total antioxidant
Correspondence capacity. Our findings suggest that NSE may have beneficial effects on thyroid gland
Gulcan Avci, Department of Biochemistry,
abnormalities owing to its antioxidant properties.
Faculty of Veterinary Medicine, University
of Afyon Kocatepe, Afyonkarahisar, Turkey. Practical applications
Email: gulcanavci@hotmail.com
Essential oils derived from Nigella sativa L. seed contain many bioactive substances
Funding information such as thymoquinone and cymene. This paper emphasizes the effect of NSE on
Afyon Kocatepe University Scientific
Research Projects Coordination Unit, Grant/ thyroid hormone abnormalities and negative oxidative state that occurs in HT and HP
Award Number: 16.VF.08 models. The present study provides evidence of a positive effect of NSE particularly
on TT3 levels in the HT and HP models. It can therefore be assumed that NSE could
be used as a supportive natural alternative source to improve thyroid hormone levels
and relieve increased oxidative stress.

KEYWORDS

antioxidative balance, black seed, hyperthyroidism, hypothyroidism, thyroid hormones

1 | I NTRO D U C TI O N gain, and constipation are typically seen in hypothyroidism, whereas

Hypothyroidism and hyperthyroidism are extremely common thy-
roid gland abnormalities worldwide. Hypothyroidism, which refers to
the underactivity of the thyroid gland, and hyperthyroidism, which
denotes the overactivity of the gland, are very common thyroid dis-
eases (Fabrizio, 2003). Bradycardia, fatigue, cold intolerance, weight
heat intolerance, weight loss, irritability, muscle weakness, diar-
rhea, and tremors are prominent in hyperthyroidism (Mounika et al.,
2013). In the treatment of hypothyroidism, thyroid hormones are
replaced by drugs such as levothyroxine. Thyrostatics, such as pro-
pylthiouracil (PTU), methimazole, and carbimazole, are used in the
treatment of hyperthyroidism. These drugs reduce thyroid hormone

J Food Biochem. 2021;00:e13801. wileyonlinelibrary.com/journal/jfbc © 2021 Wiley Periodicals LLC. | 1 of 9

https://doi.org/10.1111/jfbc.13801
2 of 9 | AVCI et al.
levels by inhibiting thyroid peroxidase enzyme, which facilitates
the iodination of thyroglobulin in the thyroid gland, or by prevent-
ing the conversion of circulating T4 to T3 (Fumarola et al., 2010). To
counter the side effects of drugs used in the treatment of diseases,
the application of natural herbal products as preventive or supple-
mentary medicines is becoming increasingly common nowadays.
Furthermore, aromatic and medicinal plants and essential oils of
these plants are viewed as natural antioxidant sources (Djeridane
et al., 2006). In particular, the fact that many synthetic drugs are
carcinogenic, expensive, and scarce has accentuated the interest in
medicinal and aromatic herbs in the medical sector.
Nigella sativa L. (NS) is a medicinal plant with a wide pharmacolog-
ical spectrum and is cultivated in many regions, such as the middle-­
eastern Mediterranean region, southern Europe, India, Pakistan,
Syria, Turkey, and Saudi Arabia (Khaled, 2009). The seeds of NS, a
plant species belonging to the Ranunculacea family, have been used
in traditional medicine since ancient times, and the plant is known
to have carminative and diuretic properties. NS is used against cold,
asthma, rheumatism, and inflammatory diseases (Randhawa & Al-­
Ghamdi, 2002; Salem, 2005). Many studies on NS seeds have shown
that the plant has antitumor, antibacterial, anti-­inflammatory, anti-
oxidant, hypoglycemic, and immunostimulant properties (Bourgou
et al., 2012; Burtis & Bucar, 2000; Randhawa & Al-­Ghamdi, 2002;
Salem, 2005). It has been stated that many of these properties are
due to thymoquinone (TQ), one of the bioactive components of
NS fixed and essential oils. In addition, the seeds contain various
essential oil components, such as dithymoquinone, thymohydro-
quinone, thymol, nigellone, carvacrol, p-­cymene, d-­limonene, and
α-­ and β-­pinene (Randhawa & Al-­Ghamdi, 2002). Oxidative stress,
known as the imbalance between the production of pro-­oxidants
(reactive oxygen species and reactive nitrogen species) and antiox-
idant defenses, is generated in both hypothyroidism and hyperthy-
roidism based on different mechanisms (Mancini et al., 2016; Resch
et al., 2002). Several studies have proven that TQ and Nigella sativa
L. oil (NSO) act as radical scavengers by suppressing oxidative stress
and protecting cells against lipid peroxidation (Farhangi et al., 2016;
Mohebbati et al. 2017). Mekkawy et al. (2020) reported that Vit C
and NSO either alone or in combination protected and restored the
structural and functional integrity of the cerebellum and the thy-
roid glands via their antioxidant properties to inhibit oxidative dam-
age induced by monosodium glutamate. It has been reported that
the age-­related increase in oxidative stress in the thyroid gland in
late adult rats almost disappeared with NSO supplementation (Zaki
et al., 2018).
Considering these beneficial effects of NS, the present study
aimed to investigate the effect of NS essential oil (NSE) administra-
tion on thyroid hormone levels and oxidant–­antioxidant balance in
rats with experimentally induced hypothyroidism and hyperthyroid-
ism. This study intended to determine the effects of NSE usage on
thyroid functions and oxidant–­antioxidant balance using the plasma
levels of thyroid hormones as well as nitric oxide (NOx), malondial-
dehyde (MDA), superoxide dismutase (SOD), and total antioxidant
capacity (TAC) in the experimental hypothyroidism model induced
by propylthiouracil (PTU) and the hyperthyroidism model induced
by L-­thyroxine (LT4).
2 | M ATE R I A L S A N D M E TH O DS
2.1 | Acquisition of the essential oils from Nigella
sativa L
NS seeds were purchased from a local herb market (Tugba company)
in Afyonkarahisar, Turkey in March 2016. They were properly
powdered using a mechanical grinder. In order to obtain essential
oil by ground NS seed, it was prepared using a Clevenger apparatus
with a solid/liquid ratio of 100 g dry NS/L and then subjected to
distillation for 3 hr. Before the beginning of distillation, 1 ml of n-­
hexane was added on the surface of water in the side arm of the
Clevenger apparatus to collect the condensed droplets of the volatile
oil, thus decreasing its partition with water to increase the yield. The
obtained essential oil was dried using anhydrous Na2SO 4 (Sigma-­
Aldrich, 238597) and kept in refrigerator at 4°C until used (Burtis &
Bucar, 2000). In the present study, NSE was administered to rats at
200 mg/kg by gastric gavage. El-­Hadiyah et al. (2003) reported that
the lethal dose (LD50) values of the intraperitoneal administration of
volatile oil, aqueous extract, and fixed oil of NS were 1.853, 3.020,
and 3.371 mg/kg, respectively. Also, the LD50 of TQ was found to
be 90.3 mg/kg. Based on the data, NS and its active constituent,
thymoquinone, were seen as a safe medicinal herb.
2.2 | NSE analysis by gas chromatography-­mass
spectrometry
Gas chromatography-­
mass spectrometry (GC/MS) analyses were
carried out using an Agilent 6890N Gas Chromatograph equipped
with a multi mode inlet (MMI; 250°C), a Agilent DB-­1MS capillary
column (30 m × 0.25 mm; film thickness 0.25 μm) and coupled with
an Agilent 5975C MS Detector, operating in the electron impact
(EI) mode at 70 eV. Transfer line temperature was set at 250°C.
The carrier gas was He (2.6 ml/min), and the oven temperature was
programmed from 50°C to 280°C at a rate of 3°C/min. The injected
volume was 2 μl and the split ratio 40:1. The identification of the
compounds was based on the comparison of their retention times
(RT) and mass spectra with those from the NIST and Wiley 2008
libraries. Relative percentages of compounds were calculated based
on the peak areas from their GC/MS chromatograms (Table 1).
2.3 | Experimental animal design
All procedures were conducted with the approval of Afyon Kocatepe
University (AKU) Animal Ethics Committee (reference number 90-­
16). A total of 48 male Wistar Albino rats weighing 180–­290 g,
5–­6 months old, were used in the present study. Experimentally
AVCI et al. | 3 of 9

TA B L E 1 Chemical composition (%) of the NSE gavage; groups with hyperthyroidism (HP), (1) control: rats were fed
a standart diet and given 1 ml 0.9% saline by gastric gavage; (2) HP:
Retention
Peak no time Components % rats were given added to daily drinking water 0.0012% LT4 (Sigma,
T2376); (3) NSE: given 200 mg/kg NSE, BW, /day, gastric gavage; and
1 2.563 Hexanal 0.052
(4) HP + NSE: rats were given added to daily drinking water 0.0012%
2 5.514 α-­Thujene 9.958
LT4 + 200 mg/kg NSE, BW/day, gastric gavage.
3 5.678 α-­Pinene 2.47
Rats were housed in the AKU Experimental Animal
4 6.033 Camphene 0.041
Implementation and Research Center. During the experimental pe-
5 6.858 Sabinene 1.167
riod, living conditions for rats were established to be 22 ± 2°C room
6 6.951 β-­Pinene 2.219 temperature and 12-­hr light/12 hr dark cycle. Standard rat feed and
7 7.629 β-­Myrcene 0.032 drinking water were given ad libitum during the 15-­day training pe-
8 8.019 α-­Phellandrene 0.039 riod. The body weight of the rats was recorded at the beginning and
9 8.339 3-­C arene 0.168 end of the 4-­week experiment.
10 8.558 α-­Terpinene 0.104
11 8.754 o-­Cymene 29.066
12 8.954 Eucalyptol (1,8-­Cineole) 0.118 2.4 | Collection of blood samples
13 9.084 α-­Limonene 1.666
After 4 weeks of study, the rats were fasted for 1 day, and then,
14 10.402 γ-­Terpinene 0.849
the animals were decapitated after 4–­5 ml of blood was collected
15 12.312 cis-­4-­methoxy thujane 0.77
with a heparin tube under deep anesthesia. Plasma were separated
16 13.433 trans-­4-­methoxy thujane 3.972
by the centrifugation of the blood samples at 3.000 rpm for 10 min
17 15.402 unidentified 0.379
at 4°C. The plasma samples obtained were stored at −20°C for
18 16.150 Terpinen-­4-­ol 0.479
the measurement of thyroid hormones and oxidant-­
antioxidant
19 17.640 4.5-­Epoxy-­1-­Isopropyl-­4-­ 0.617 parameters.
methyl-­1-­c yclohexene
20 19.002 2-­methyl-­3-­phenyl-­propanal 0.197
21 19.258 Carvone 0.074 2.5 | Measurements of thyroid hormones and
22 19.850 Thymoquinone 43.745 oxidant-­antioxidant parameters
23 21.854 β-­Ocimene 0.043
24 22.147 Anethole 0.041 Plasma free triiodothyronine (fT3) and free tetraiodothyronine (fT4)
25 22.639 Bornyl acetate 0.087 levels were measured by electrochemiluminescence immunoassay
26 23.850 Carvacrol 0.694 method (Roche, 03,051,986 190 and 11,731,297 122, respectively)

27 24.081 2,4-­Decadienal 0.079 with autoanalyzer using a commercial kit. Plasma total triiodothyronine
(TT3) and total tetraiodothyronine (TT4; Biovander, RD291001200R)
28 27.015 α-­Longipinene 0.126
were determined by the enzyme-­linked immunosorbent assay (ELISA)
29 28.689 Copaene 0.019
method in accordance with the procedure inserts of commercial kits.
30 29.647 Seychellene 0.026
Plasma nitric oxide (NOx; Miranda et al., 2001), malondialdehyde
31 29.918 Longifolene 0.621
(MDA; Draper & Hadley, 1990), and superoxide dismutase (SOD; Sun
32 30.891 Caryophyllene 0.048
et al., 1998) levels were performed in accordance with the mentioned
33 41.086 Isoapiole 0.034 procedure. Plasma total antioxidant capacity (TAC) levels were
Total 100.000 determined by ELISA kits (antioxidant assay kit, item no. 709,001,
Note: Bold text indicates significant values, and they are the most Cayman Chemical Company).
intense substances in terms of content.

hypothyroidism (Moulakakis et al., 2008) and hyperthyroidism 2.6 | Statistical analysis

(Subudhi et al., 2008) were performed in eight groups, two control
and six experimental groups, with six rats in each group as follows:
groups with hypothyroidism (HT), (1) control: rats were fed a
standart diet and given 1 ml 0.9% saline by gastric gavage; (2) HT:
rats were given added to daily drinking water 0.05% PTU (Sigma,
P3755); (3) NSE: given 200 mg/kg NSE, body weight (BW)/day,
gastric gavage; and (4) HT + NSE: rats were given added to daily
drinking water 0.05% PTU + 200 mg/kg BW, NSE/day, gastric
A statistics-­package program (Statistical Package for the Social
Sciences, SPSS) was used for the analysis of the data. The means
that the data were given as mean ± standard error values. Normality
tests were conducted, and the statistical differences among groups
were analyzed through One way ANOVA. Duncan test was used as
a posttest. Statistical significance of all data was determined as p <
.001, p < .01, p < .05.
4 of 9 | AVCI et al.

3 | R E S U LT S 4 | D I S CU S S I O N

3.1 | Chemical composition of the Nigella sativa L.
essential oil
As given in Table 1, the main constituents of NSE have been identified
as TQ (43.745%) and o-­cymene (29.066%).
3.2 | Changes of body weight
As shown in Table 2, no change was found in the NSE group compared
to the control group and in the HT + NSE group compared to the HT
group in terms of body weight at the end of the trial period. There
was no significant change in body weight in the hyperthyroidism
groups compared to the control group.
3.3 | Thyroid hormone and oxidant-­
antioxidant levels
As seen in Table 3, it was determined that the fT3, fT4 (p < .001),
and TT3 (p < .01) levels decreased significantly in the HT group
compared to the controls. Simultaneously, it was found that NOx
and MDA values increased significantly (p < .01) in the HT group,
whereas plasma TAC level decreased (p < .001). Compared to
the control group, the values of all parameters in the NSE group
were statistically insignificant. When evaluated in terms of
thyroid hormone levels, it was found that TT3 level increased
significantly (p < .01) and NOx level decreased significantly (p < .01)
in the HT + NSE group compared to the HT group. As indicated
in Table 4, when the control group was compared with the HP
group, it was observed that the fT3, fT4 (p < .001), and TT3 (p <
.05) levels increased significantly. In addition, it was noted that
plasma TAC level decreased significantly (p < .01) and MDA level
increased significantly (p < .05) in the HP group. Compared to
the control group, the values of all parameters in the NSE group
were statistically insignificant. In terms of thyroid hormones, when
the HP + NSE group was compared with the HP group, it was found
that the TT3 level decreased significantly (p < .05) and the TAC level
increased significantly (p < .01).
While medicinal plants are used as natural alternatives to support
treatment in various diseases, they serve as the only effective
treatment in some cases (Salem, 2005). NS, an aromatic plant, is
rich in phenolic compounds. These compounds have antioxidant
properties that have traditionally been used for centuries in the
treatment of various diseases such as fever, headache, asthma,
anxiety, and stroke (Randhawa & Al-­Ghamdi, 2002). Kıralan (2014)
reported that the essential oil content in NS seeds is around 0.5%–­
1.4%. In a previous study, the components found in NSE have been
reported to be TQ, dithymoquinone, thymohydroquinone, thymol,
p-­cymene, d-­limonene, α-­ and β-­pinene, trans-­anethole, carvacrol,
and nigellone (Ali & Blunden, 2003). The main compounds that
we detected in NSE in the present study were TQ (43.74%) and o-­
cymene (29.06%). TQ, one of the active compounds found in NS in
the present study, has many biological activities such as antioxidant,
anti-­inflammatory, antimicrobial, and anticancer activities (Ali &
Blunden, 2003; Salem, 2005). Another active compound, cymene,
is an organic aromatic compound that is found in the volatile oils
of more than 100 plants. Studies have reported that cymene
exhibits antioxidant, analgesic, and anti-­inflammatory properties in
mice (Bonjardim et al., 2012; Oliveira et al., 2015). In other studies,
the levels of NSE components in Turkey were reported to be TQ
(44.77%) and p-­cymene (28.62%) by Kıralan (2014) and TQ (45.78%)
and p-­cymene (29.45%) by Erdogan et al. (2020), which support our
results. These small differences in the results in terms of the major
compounds could be attributed to factors such as variations in the
geographical conditions in which the plant grows and the method
of analysis.
The thyroid hormones are involved in the regulation of energy
expenditure and body weight. It is known that the lack of these hor-
mones could reduce the energy expenditure and increase the fat
mass (Syed et al., 1999). The significant decrease found in the aver-
age body weight of rats in the HT group compared to the controls
at the end of the experimental period in our study was consistent
with Kandır (2015)’s report that the average body weight and feed
consumption in rats with hypothyroidism decreased significantly.
Similarly, other studies reporting a decrease in metabolizable energy
intake and consumption (Iossa et al., 2001) and a dose-­dependent
decrease in body weight and food intake (Nambiar et al., 2014) in

TA B L E 2 Changes in body weight of rats with experimental induced hypothyroidism and hyperthyroidism (x̄ ± SEM)

Parameter Control HT NSE HT + NSE p

a b a b
Body weight (g) 290.66 ± 5.49 212.00 ± 3.89 267.16 ± 11.37 186.66 ± 12.45 .001
Control HP NSE HP + NSE p
Body weight (g) 235.00 ± 9.03 185.40 ± 13.26 232.66 ± 34.00 215.50 ± 18.332 .396

Note: Hypothyroidism (HT), a group containing 0.05% propylthiouracil (PTU) for 4 weeks in drinking water. NSE, a group where the essential oil of
Nigella sativa L. is administered orally at dose of 200 mg/kg for 4 weeks. HT + NSE, a group where 0.05% PTU was added to drinking water and also
essential oil of Nigella sativa L. was administered orally at dose of 200 mg/kg. Hyperthyroidism (HP), a group containing 0.0012% L-­thyroxine (LT4)
for 4 weeks in drinking water. NSE, A group where the essential oil of Nigella sativa L. are administered orally at dose of 200 mg/kg for 4 weeks.
HP + NSE, a group where 0.0012% LT4 was added to drinking water and also essential oil of Nigella sativa L. was administered orally at dose of
200 mg/kg. Different letters in the same line represent the statistically significant difference between the mean ± SEM values (p < .05).
AVCI et al. | 5 of 9

TA B L E 3 Plasma thyroid hormones and oxidant-­antioxidant parameters in rats with experimental hypothyroidism (x̄ ± SEM)

Parameter Control HT NSE HT + NSE p

a b a b
fT3 (pg/ml) 2.26 ± 0.18 1.06 ± 0.07 2.64 ± 0.13 0.92 ± 0.06 .000
fT4 (ng/dl) 1.90 ± 0.13a 0.10 ± 0.02b 1.84 ± 0.14a 0.19 ± 0.02b .000
ab b ab ab
TT4 (ng/ml) 53.91 ± 2.321 44.96 ± 4.13 49.79 ± 1.27 45.87 ± 2.93 .116
TT3 (pg/ml) 781.05 ± 15.98b 610.82 ± 23.95a 806.15 ± 26.33b 769.49 ± 51.13b .003
b a b b
NOx (µmol/L) 21.22 ± 0.888 28.79 ± 0.769 24.19 ± 1.41 23.99 ± 1.85 .010
MDA (nmol/ml) 2.61 ± 0.198b 3.62 ± 0.220a 2.50 ± 0.241b 3.48 ± 0.225a .003
ab b a b
SOD (U/ml) 1.22 ± 0.071 0.96 ± 0.066 1.42 ± 0.122 1.06 ± 0.092 .015
TAC (mmol/L) 0.460 ± 0.0279ab 0.338 ± 0.0162c 0.493 ± 0.0209a 0.400 ± 0.0148bc .000

Note: Hypothyroidism (HT), a group containing 0.05% propylthiouracil (PTU) for 4 weeks in drinking water. NSE, a group where the essential oil of
Nigella sativa L. is administered orally at dose of 200 mg/kg for 4 weeks. HT + NSE, a group where 0.05% PTU was added to drinking water and also
essential oil of Nigella sativa L. was administered orally at dose of 200 mg/kg. Different letters in the same line represent the statistically significant
difference between the mean ± SEM values (p < .001, p < .01, p < .05).

TA B L E 4 Plasma thyroid hormones and oxidant-­antioxidant parameters in rats with experimental hyperthyroidism (x̄ ± SEM)

Parameter Control HP NSE HP + NSE p

fT3 (pg/ml) 2.87 ± 0.07b 11.12 ± 1.91a 2.73 ± 0.15b 13.73 ± 2.27a .000
b a b a
fT4 (ng/dl) 2.10 ± 0.10 7.77 ± 0.00 2.01 ± 0.10 7.77 ± 0.00 .000
TT4 (ng/ml) 52.16 ± 2.92 62.32 ± 2.96 54.23 ± 4.28 61.18 ± 2.94 .143
b a b b
TT3 (pg/ml) 782.63 ± 37.65 928.88 ± 17.47 806.49 ± 41.84 773.77 ± 51.31 .050
NOx (µmol/L) 21.59 ± 0.87 16.83 ± 1.16 21.02 ± 0.912 18.99 ± 2.48 .110
b a b ab
MDA (nmol/ml) 2.85 ± 0.175 3.43 ± 0.195 2.66 ± 0.135 2.95 ± 0.159 .026
SOD (U/ml) 1.31 ± 0.12048 1.18 ± 0.12306 1.50 ± 0.12593 1.15 ± 0.09522 .158
TAC (mmol/L) 0.452 ± 0.018a 0.318 ± 0.035b 0.457 ± 0.019a 0.442 ± 0.0222a .002

Note: Hyperthyroidism (HP), a group containing 0.0012% L-­thyroxine (LT4) for 4 weeks in drinking water. NSE, a group where the essential oil of
Nigella sativa L. is administered orally at dose of 200 mg/kg for 4 weeks. HP + NSE, a group where 0.0012% LT4 was added to drinking water and also
essential oil of Nigella sativa L. was administered orally at dose of 200 mg/kg. Different letters in the same line represent the statistically significant
difference between the mean ± SEM values (p < .001, p < .01, p < .05).

rats with PTU-­induced hypothyroidism support our results. In addi-
tion, Soukup et al. (2001) stated that the body weight gain per week
was very low during the experimental period in hypothyroid rats and
that their body growth was practically arrested. In the light of this
information, the body weight reduction of the rats in the HT group
may be related to the decrease in feed intake and basal metabolism.
Furthermore, low feed intake might have partly reduced the water
intake.
In studies where TQ, one of the components of NS seeds,
was administered orally to rats at a dose of 80 mg/kg (Pari &
Sankaranarayanan, 2009) and 10 mg/kg (Badary et al., 2000), the
body weight of the rats did not change. Similarly, in both the hy-
pothyroidism and hyperthyroidism models of the present study,
the changes in body weights were found to be insignificant in the
groups that were administered only NSE when compared to the
respective controls. On the other hand, Zaoui et al. (2002) un-
covered that the administration of NS fixed oil (1 ml kg−1 day−1,
12 weeks) resulted in a significant reduction in the body weight
from the 6th week, and this could be due to the effect of TQ on
the food intake. Similarly, it has been documented that TQ admin-
istration (50 mg/kg) resulted in decreased body weight at the end
of the 6th week in rats fed with a fatty diet (Gullu & Avci, 2013).
Considering the use of NSO/TQ in the abovementioned studies,
the fact that the decrease in body weight seen in the HT + NSE
group was not significant compared to that in the HT group in our
study may be due to differences in the trial period or the use of
NSE. Studies have reported that LT4 administration causes loss
of body weight (Shinohara et al., 2000) and that basal metabo-
lism is accelerated in hyperthyroidism, leading to increased en-
ergy consumption and decreased body fat mass, thereby leading
to decreased body weight (Greenlund et al., 2008). The changes
observed in all other groups in the hyperthyroidism models were
insignificant when compared to the controls, which may be due to
the short trial period.
In the present study, it was found that fT3, fT4, and TT3 levels de-
creased significantly in the HT group (no change in TT4), whereas fT3,
fT4, and TT3 levels significantly increased in the HP group. According
to the results obtained, the decrease in thyroid hormone levels fol-
lowing the administration of PTU in the HT group and the increase
in thyroid hormone levels following the administration of LT4 in the
HP group prove that experimental hypothyroidism and hyperthy-
roidism were achieved in both models of the study. It is known that
6 of 9 | AVCI et al.
PTU, which used to induce experimental hypothyroidism, exerts its
effect by inhibiting the ionization of monoiodotyrosine (binding of
iodine to tyrosine) and oxidation of iodine (by inhibiting tyrosine
peroxidase, which transforms I− to I2) and by preventing the periph-
eral conversion of T4 to T3 (by inhibiting type-­I deiodinase; Fumarola
et al., 2010). LT4, which is used to induce experimental hyperthy-
roidism, is a synthetic thyroxine hormone. These findings with re-
gard to the thyroid hormones are consistent with studies in which
experimental hypothyroidism and hyperthyroidism were induced in
rats (Moulakakis et al., 2008; Subudhi et al., 2008). In both models of
the present study, it was found that the changes in the levels of thy-
roid hormones and oxidant–­antioxidant parameters in the NSE group
were not statistically significant when compared to the control
group. Although there is not enough data on the subject, Jasima et al.
(2016) have reported that administration of NSO (1 ml kg−1 day−1) to
rats increased TT3 and TT4 levels, whereas Gullu and Avci (2013)
reported that administration of TQ (50 mg kg−1 day−1) decreased
TT4 levels significantly, with insignificant decrease in other thyroid
hormone levels. Taking our findings into account, NSE administra-
tion did not adversely affect the thyroid hormones and the oxidant–­
antioxidant balance.
In the present study, it was found that TT3 level was higher in the
HT + NSE group when compared to the HT group and the changes in
the levels of other thyroid hormones were insignificant. In the liter-
atüre search, no study was found in which any NSE was used to treat
hypothyroidism and hyperthyroidism. However, Khalawi et al. (2013)
have opined that NS (400 mg/kg) administration increased serum
T3 and T4 levels, decreased TSH levels, and reversed hyperplastic
changes in the thyroid gland in rats with hypothyroidism. In another
study, Mohebbati et al. (2017) have observed that in rats with PTU-­
induced hypothyroidism, serum T4 level decreased in the PTU group
compared to controls, whereas it increased in all groups adminis-
tered with NS (100, 200, and 400 mg/kg). The highest increase was
observed in the 400 mg/kg NS group. Similarly, Meral et al. (2003)
have alluded that low fT3 levels in rabbits with diabetes increased
with NS administration and approached normal levels; hence, NS
may alleviate thyroid hormone metabolic disorders caused by dia-
betes. In patients with Hashimoto’s thyroiditis, in which hypothy-
roidism is most common, NS administration has been reported to
increase serum T3 levels and decrease TSH and anti-­thyroid peroxi-
dase (anti-­TPO) levels (Farhangi et al., 2016). Shariatifar et al. (2014)
have reported that the administration of hydroalcoholic extract (50,
100, and 200 mg/kg) of NS in rats increases serum T3 and T4 levels
while decreasing the TSH levels. Considering the studies mentioned
above, it could be seen that both NS and NSO administrations exert
positive effects on thyroid abnormalities owing to their antioxi-
dant properties. Similarly, the results obtained in our study suggest
that NSE has improvement effects on thyroid hormones, especially
due to increased TT3. It was determined that the TT3 level in the
HP + NSE group decreased significantly, except for the other thyroid
hormones, and approached the control levels compared to the HP
group. However, there are not enough studies on the subject with
which we can compare our results.
Thyroid hormones are closely associated with oxidative me-
tabolism, regulation of antioxidant enzyme levels, and the syn-
thesis of antioxidants. Antioxidant enzymes act by scavenging
the free radicals and converting them to less harmful molecules,
thus protecting the body from their damaging effects. It is well
known that reducing antioxidant levels in the circulation leads
to oxidative stress (Venditti et al., 1997). The researchers stated
that free radical formation decreases and thus oxidative stress
is lowered due to the decrease in lipid peroxidation and as a re-
sult of the slowing of the basal metabolism in the hypothyroidism
(Araujo et al., 2011; Harper & Seifert, 2008). When our results
were evaluated in terms of antioxidant metabolism, it was found
that NO x and MDA levels increased significantly in the HT group
compared to the control group, whereas TAC level decreased. In
our experimental model, the decrease in TAC and the increase in
NOx and MDA in the HT group may have been caused by distur-
bances in the antioxidant synthesis capacity (Reddy et al., 2013).
Considering the above information, our findings support this in-
formation. In the same study, although SOD and NO x levels did
not change in the HP group compared to the control group, TAC
level decreased, and MDA level increased significantly. Mogulkoc
et al. (2005) has stated that hyperthyroidism induced by LT4 acti-
vates both MDA and reduced glutation (GSH) in cerebral, hepatic,
and cardiac tissues. Kandır (2015) has reported that plasma MDA
level of rats increased in both experimentally induced hypothy-
roidism and hyperthyroidism groups compared to the control
group, and their results support our findings. In fact, there are
different reports in terms of NO x levels in hypothyroidism and hy-
perthyroidism. Kandır (2015) has reported that plasma NO x levels
were not altered in the hypothyroidism group compared to the
control group; however, these levels decreased in the hyperthy-
roidism group, and this report was not consistent with the results
of our study. There are also reports that plasma NO x levels are ei-
ther not affected (Hermenegildo et al., 2002) or increase (Arikan
et al., 2007) in hypothyroidism. In fact, Verma et al. (2015) has
reported that serum NO x levels increased in people with hypo-
thyroidism and decreased in people with hyperthyroidism. The
results of the corresponding levels of NO x in the experimental HT
group in the present study support this report. Accordingly, the
increase in lipid peroxidation occurring in hyperthyroidism could
be explained by the inadequate antioxidant response against
free radicals released from mitochondrial respiratory chain due
to the acceleration of the basal metabolism rate. In contrast, it
has been stated that plasma NO x levels decreased in people with
hyperthyroidism (Arikan et al., 2007; Hermenegildo et al., 2002);
however, the decrease in NO x level in the hyperthyroidism group
in our study was not significant. In our experimental model, the
decrease in TAC and the increase in MDA in the HP group may
have been caused by increased free radicals due to the acceler-
ated basal metabolism. Disturbances of the oxidant–­a ntioxidant
balance resulting from the increased production of free radicals
lead to oxidative damage of biological membranes and thus in-
crease lipid peroxidation (Halliwell & Guteridge, 1989). We found
AVCI et al.
that NSE administered in both hypothyroidism and hyperthyroid-
ism models did not cause any changes in SOD and MDA and only
increased TAC in the HP + NSE group compared to the control
group. Our findings can be explained by the fact that the increase
in TAC in the group treated with NSE may be due to the positive
effects of NSE on the antioxidant system. The main ingredients
of NSE, such as TQ (Farhangi et al., 2016) and cymene (Oliveira
et al., 2015), are also known to enhance the antioxidant activity.
In addition, polyphenolic compounds and flavonoids with many
hydroxyl groups in NSE may be responsible for the increase in
the total antioxidant capacity due to their free radical scavenging
activities.
In a study conducted by Mohebbati et al. (2017), when the kid-
ney tissue was examined in rats with PTU-­induced hypothyroid-
ism, it was determined that the MDA level increased. On the other
hand, catalase and SOD activity decreased in the PTU group com-
pared to the levels observed in the controls. These values were
reversed in the NS-­t reated groups (200 and 400 mg/kg), and the
increase was significant. Based on the results of this study, it could
be stated that hydroalcoholic NS extract has protective effects on
oxidative kidney damage observed in hypothyroidism caused by
PTU. Considering the report that both fixed oil and essential oil of
NS have antiradical and antioxidant properties (Randhawa & Al-­
Ghamdi, 2002), it is believed that NSE exerts positive effects via
TT 3 in both hypothyroidism and hyperthyroidism. These effects
are thought to occur because of the antioxidant properties of TQ
and cymene, which are found in high amounts in NSE (Oliveira
et al., 2015; Salem, 2005). It can thus be interpreted that these
two agents help in lowering the lipid peroxidation caused by the
increased oxidative stress in the thyroid gland cells in hypothyroid-
ism and hyperthyroidism. Besides, these agents increase TAC, es-
pecially in hyperthyroidism.
5 | CO N C LU S I O N
In conclusion, the study has asserted that NSE administration in
rats increased TT3 and decreased NOx in HT, whereas it decreased
TT3 and increased TAC in HP. Considering these results, it could be
concluded that NSE has a favorable impact on the total antioxidant
capacity and has protective effects in both HT and HP models. As a
result, it can be stated that NSE plays a protective and supportive
role in thyroid abnormalities.
AC K N OW L E D G M E N T S
This work was supported by Afyon Kocatepe University Scientific
Research Projects Coordination Unit (Project number: 16.VF.08).
This study was presented as a poster presentation at Central
Bohemia University International Conference Innovations in Science
and Education March 21–­23, 2018, Prague, Czech Republic.
C O N FL I C T O F I N T E R E S T
The authors declared that they have no conflict of interest.
| 7 of 9
AU T H O R C O N T R I B U T I O N S
Funding acquisition; Methodology; Project administration; Supervision;
Validation; Writing-­original draft; Writing-­review & editing: Gulcan Avci.
Investigation; Resources; Supervision: Elmas Ulutas. Conceptualization;
Investigation; Resources; Visualization: Vural Özdemir. Data curation;
Investigation; Resources; Supervision: Ibrahim Kivrak. Conceptualization;
Data curation; Formal analysis; Investigation; Resources; Validation;
Visualization; Writing-­review & editing: Aziz Bulbul.
ORCID
Gulcan Avci https://orcid.org/0000-0001-8767-4507
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