Download A Histological Study on the Effect of Diclofenac Sodium

Egypt. J. Histol. Vol. 33, No. 2, June, 2010: 213 - 223
(ISSN: 1110 - 0559)
Original Article
A Histological Study on the Effect of Diclofenac Sodium (Declophen)
Administration on Thyroid Follicular Cells of Albino Rats
Nadia M. El-Rouby
Histology Department, Faculty of Medicine, Cairo University
ABSTRACT
Introduction: Diclofenac Sodium (declophen) is a nonsteroidal anti-inflammatory drug recently categorized as thyroid
antagonist.
Aim of the Work: To determine the histo-physiological changes that might occur in the thyroid follicular cells after
declophen administration and evaluate their reversibility.
Materials and Methods: Thirty adult male albino rats were randomly divided into three groups; Group I (Control
group), Group II received declophen (8 mg/kg /day) for one week then sacrificed. Group III received declophen as
group II and then sacrificed one week after drug withdrawal. Blood samples were collected to estimate T3, T4 and TSH.
Thyroid glands were processed for microscopic examination. Mean follicular diameter and mean height of thyrocytes
were morphometerically evaluated and statistically analyzed.
Results: Declophen significantly decreased T3 level which did not return back to normal in group III. Histologically,
group II showed considerable light microscopic changes as focal distension of follicles that were lined with flattened
epithelium. Their colloid revealed minimal peripheral vacuolations. Some follicles showed shedded epithelial lining.
Others appeared irregular with disrupted epithelium. That epithelium had irregular dark pyknotic nuclei and vacuolated
cytoplasm. Some follicles had apparently little colloid. Ultrastructural changes included dilated rER with electron lucid
material and many darkly stained lysosomes. Group III showed some changes compared with Control. Statistically, mean
follicular diameter of group II increased significantly while mean epithelial height decreased significantly compared to
Control. Group III revealed neither significant increase in mean follicular diameter or decrease in epithelial height in
comparison with the control.
Conclusion: Declophen has toxic effect on the thyroid follicular cells which was incompletely recovered. So it should
be discontinued prior to evaluation of thyroid functions.
Key Words: Declophen, follicular cells, thyroid gland,
Corresponding Author: Nadia M. El-Rouby
rats.
Tel.: 0111675532 E-mail: [email protected]
INTRODUCTION
Many factors including drugs can influence thyroid
function in experimental animals and humans. Studies
in humans reported significant effects of non steroidal
anti-inflammatory drugs (NSAIDs) on thyroid tests,
which can lead to misinterpretation of the result and
inappropriate therapeutic decision1-3.
and anti-pyretic actions. It is commonly used as a pain
killer in many cases including rheumatoid arthritis,
dysmenorrhea, artheralgia, tooth ache and postoperative
pain7. Sometimes the drug may be used in high doses due
to severe agonizing pain or by mistake.
As NSAIDs are used more and more frequently in
human, it is important to know to what extent they can
influence the thyroid gland. Reviewing the literature
showed that the influence of NSAIDs, including
declophen, on the concentration of thyroid hormones
has been established. However, there have been very few
previous studies demonstrating the effect of declophen
on the histological architecture of thyroid gland8. So
the objective of the present study was to evaluate the
declophen-induced alterations in the histological structure
and function of the thyroid follicular cells. It was also
aimed to evaluate the reversibility of such changes.
The role of NSAIDs in causing thyroid dysfunction
is quite often forgotten. These drugs might influence
thyroid hormone homeostasis at any level from their
synthesis, secretion, and transport or end-organ action
as it might interfere with thyroid hormone binding
sites (competitive inhibition). They might also alter
the synthesis and secretion of thyrotropin (Thyroid
Stimulating Hormone: TSH)4,5. Some cases, however,
showed clinically apparent thyroid disease6.
Diclofenac Sodium (Declophen) is a potent NSAID.
It has anti-rheumatic, anti-inflammatory, analgesic
20 (1185-2010)
213
Nadia M. El-Rouby
were fixed in 10% formol saline and processed
for paraffin blocks. Sections of 5 micrometer
thickness were cut and stained with hematoxylin
and eosin (H&E).
MATERIALS AND METHODS
A- Drug:
The drug used was Diclofenac Sodium (declophen),
presented as ampoules of 75 mg/3ml (Pharco
Pharmaceutical, Alexandria, Egypt). The dose used for
rats was 8 mg/kg body weight9. The drug was given daily
for a week by intramuscular (I.M) route. The control rats
were given physiological saline I.M. in the same volume
daily.
2. For Transmission Electron Microscopy (TEM):
Thyroid tissues were rinsed in phosphate buffer
(PH 7.4) then fixed in 2.5% gluteraldehyde, post
fixed in 1% osmium tetroxide and embedded in
epoxy resin. Semithin sections of 1 micrometer
thickness were cut on an LKB ultramicrotome
and stained with toluidine blue. Ultrathin
sections were stained with uranyl acetate and
lead citrate10 to be examined under Joel EM
100 S TEM using an accelerated voltage of 60
KV.
B- Animals:
Thirty adult male albino rats were used in this study.
They were obtained from Kaser El -Aini animal house.
Their weights ranged between 150-200 g. They were
kept under good hygienic conditions, fed ad libitum and
allowed free water supply. Rats were divided randomly
into three equal groups as follows:
Morphometric Analysis of Both Thyroid Follicular
Diameter and Follicular Epithelium Height:
Leica Qwin 500 Ltd, image analysis system was used
to determine the diameter and the follicular epithelium
height of thyroid follicles. The two parameters were
measured in micrometers using low power (X 100) and
high power (X 400) magnification, respectively. The
internal follicular diameters of 50 follicles, in 10 random
fields, of the thyroid gland of each rat of the different
groups were measured. They were measured on diagonal
axes. The mean of these two readings was then taken
as the internal diameter of the follicle. To measure the
epithelial height, 50 follicles from each rat in 10 random
fields were chosen. The epithelial height was measured
at two points on the same axis of each follicle. The mean
of these two readings was then taken as the epithelial
height. The mean value and Standard Deviation (SD) of
the data obtained for each group were calculated.
Group I: (Control Group):
Rats were given physiological saline daily in a similar
manner to other groups. Five rats were sacrificed with
animals of group II and the other five rats were sacrificed
with animals of group III.
Group II: (Experimental Group):
Rats were given declophen as an I.M. daily dose of 8
mg/kg body weight 9 for one week. After that week, the
rats were sacrificed.
Group III: (Recovery Group):
Rats received declophen as in group II. The rats were
kept alive for another week without treatment. On the
15th day of the beginning of the experiment these rats
were sacrificed.
Statistical Analysis:
Statistical analysis was performed on IBM/PC
using SPSS (Version 11) / PC program. Comparison of
significance between the different groups was made using
student “T” test11. Results were considered significant
when probability (p) ≤ 0.05.
Methods:
At the end of the experimental period, the animals
were sacrificed under ether anaesthesia. Blood samples,
from all animals, were collected for hormonal assay.
Then, the thyroid gland of each rat was excised.
RESULTS
During the experiment, three rats died; two from the
experimental group (Group II) and one from the recovery
group (Group III).
Hormonal Assay:
Hormonal assay was carried out at the Clinical
Pathology Department, Faculty of Medicine, Cairo
University, to determine the serum levels of T3, T4 and
TSH.
Hormonal Assay:
It was clear that T3 was significantly decreased in
both the experimental and recovery groups (P< 0.05)
compared to the control. On the other hand, there was
no significant change of both T4 and TSH (P >0.05)
compared to the control (Table 1 and Chart 1).
Histological Study:
1. For Light Microscopy: The thyroid tissues
214
A Histological Study on the Effect of Diclofenac Sodium (Declophen) Administration on Thyroid Follicular Cells of Albino Rats
Table 1: The mean values of T3 in µg/dL, T4 in µg/dL and TSH in µU/mL ± SD in different groups:
Item
Control group
Experimental group
Recovery group
T3: Mean ±SD
“T” test
3.9685 ±.09540
2.7100 ±.07071
* P1< 0.05
3.0990 ±.02767
* P2< 0.05
T4: Mean ±SD
“T” test
2.6270 ±.02830
2.6150 ±.02550
P1 >0.05
2.6000 ±.02944
P2 >0.05
0.0118 ± 0.00301
0.0137 ± 0.00295
P1 >0.05
0.0120 ±.00267
P2 >0.05
TSH: Mean ±SD
“T” test
P1 = between the experimental group and control group.
P2= between the recovery group and control group.
* Significant P value.
was interrupted (Fig. 4-B). The lining follicular cells of
the damaged follicles had darkly stained irregular and
pyknotic nuclei. The cytoplasm of the affected thyrocytes
was vacuolated (Figs. 5-A and B). The follicles showed
variable density of colloid staining. Mast cells were
frequently seen in the interstitial tissue close to blood
vessels (Fig. 6). Electron microscopy revealed that the
nuclei of the affected follicular cells appeared indented
and hetrochromatic with dilatation of the peri-nuclear
space. The cisternae of rER were dilated and contained
amorphous lightly stained material (Figs. 7 and 8).
The cytoplasm showed many darkly stained lysosomes
and prominent Golgi complex (Fig. 9). The distended
follicles were lined with flattened thyrocytes that had flat
nuclei (Fig. 10).
Chart 1: The mean value of T3, T4 & TSH levels.
Group III (Recovery Group): The thyroid follicles
of rats of this group started to regain their activity and
normal appearance but patchy in distribution. These
follicles were lined with cubical epithelium and showed
peripheral vacuolations of their colloid content. Many
distended follicles were still seen. These follicles
were lined with flattened thyrocytes. Their colloid had
minimal peripheral vacuolations. Also, involuted and
distorted follicles persisted (Fig. 11). The follicular cells
had vesicular nuclei. Some mast cells with darkly stained
granules were seen in the surrounding connective tissue
beside the blood vessels (Fig.12). Electron microscopy
revealed that the follicular cells appeared more or less
normal compared with the control group. They had
more or less regular euchromatic nuclei with prominent
nucleoli. The cytoplasm had more or less normal rER
cisternae but many vacuoles and many darkly stained
lysosomes were still seen in some follicular cells
(Figs. 13 and 14). Some follicular cells showed sings of
regeneration as they were binucleated (Fig. 15).
Histological Results:
Group I (Control Group): The thyroid gland of
control rats was composed of follicles lined with a
single layer of cuboidal follicular cells, and filled with
acidophilic homogenous colloid having peripheral
vacuolations (Figs. 1-A and B). The thyroid follicles
were surrounded by vascular connective tissue stroma
(Figs. 1-A and 2-A).The follicular cells had vesicular
nuclei and prominent nucleoli (Fig. 2-A). Electron
microscopy revealed that the follicular cells had
euchromatic nuclei, rough endoplasmic reticulum and
mitochondria. The apical cytoplasm revealed secretory
vesicles. The colloid showed peripheral vacuolations in
the form of electron lucid areas (Fig. 2-B).
Group II (Experimental group): Examination
of the thyroid gland of rats of the experimental group
revealed focal markedly distended thyroid follicles.
These follicles were lined mostly with flattened epithelial
cells that had flattened nuclei. Their colloid showed
nearly absent peripheral vacuolations (Fig. 3-A). Some
follicles revealed shedded cells. The interstitial tissue had
dilated blood vessels (Fig. 3-B). The thyroids of some
animals revealed focal involuted follicles with minimal
amount of colloid (Fig. 4-A). The wall of some follicles
Morphometric Study and Statistical Analysis:
The mean diameter of thyroid follicles of rats of the
experimental group was significantly increased (p1< 0.05)
compared to the control group while the mean follicular
diameter of thyroid follicles of the recovery group was
insignificantly increased (p 2 >0.05) in comparison with
the control group (Table 2 and Chart 2).
215
Nadia M. El-Rouby
Table 2: The mean follicular diameter in µm ±SD of thyroid follicles of different animal groups:
Mean follicular diameter in µm
“T” test
Control group
Experimental group
Recovery group
35.7±9.6
51.9±13
* P1< 0.05
36.6±9.9
P2>0.05
P1 = between the experimental group and control group.
P2= between the recovery group and control group.
* Significant P value.
Chart 2: The mean follicular diameter of thyroid follicles of the different groups.
The mean epithelial height of thyroid follicles of
the experimental group was significantly decreased
(p1< 0.05) compared to the control group while the mean
epithelial height of the recovery group was insignificantly
decreased ( p2 >0.05) compared with the control group
(Table 3 and Chart 3).
Table 3: The mean value of epithelial height of thyroid follicles in µm ± SD in different groups:
Mean epithelial height in µm
“T” test
Control group
Experimental group
Recovery group
9.2±1.6
6.8±1.4
* P1< 0.05
8.9±1.9
P2>0.05
P1 = between the experimental group and control group.
P2= between the recovery group and control group.
* Significant P value.
Chart 3: The mean epithelial height of thyroid follicles in the different groups.
216
A Histological Study on the Effect of Diclofenac Sodium (Declophen) Administration on Thyroid Follicular Cells of Albino Rats
Fig. 1: Photomicrographs of sections in the thyroid gland of a control rat
(group I) showing:
a. Normal thyroid follicles lined with simple cuboidal epithelium and
filled with colloid (C). Note blood vessels (BV) in the surrounding
connective tissue stroma.
H&E X 200.
b. The colloid (C) has peripheral vacuolations (V).
H & E X 400.
Fig. 2: Photomicrographs of:
a. Semithin section in the thyroid gland of a control rat. It shows that
the follicles are of variable sizes and lined with cuboidal follicular
cells that have vesicular nuclei (N). The follicles are filled with
colloid (C). Note that the stroma has many blood capillaries.
Toluidine blue X 1000.
b. TEM section of a control rat thyroid showing a part of thyrocyte.
The colloid (C) shows two electron lucid areas (V) of peripheral
colloid vaculations. Cisternae of RER and mitochondria (m) are
scattered throughout the cytoplasm. Many secretory vesicles
(S) in the apical cytoplasm and part of the nucleus (N) are seen.
Orig mag X 2000.
217
Fig. 3: Photomicrographs of sections in the thyroid gland of rats from the
experimental group (Group II):
a. Reveals markedly distended thyroid follicles. They are lined mainly
by flat cells with flat nuclei (arrow) and few low cuboidal cells with
apparently rounded nuclei (arrow heads). The colloid (C) shows
apparently no peripheral vacuolations.
H&E X 400.
b. Shows that some follicles have shedded epithelial lining (*). The
thyrocytes of most of the follicles have vacuolated cytoplasm
(arrow). Note the dilated blood vessel (b.v).
H&E X 400.
Fig. 4: Photomicrographs of sections in the thyroid gland of rats from the
experimental group (Group II):
a. Shows thyroid follicles of variable activity as some follicles are
markedly distended (1) and other follicles appear involuted (2).
These follicles have minimal amount of colloid.
H&E X 200.
b. Reveals a follicle with interrupted follicular wall (arrow). The
adjacent follicles showing thyrocytes with vacuolated cytoplasm
(curved arrow).
H&E X 400.
Nadia M. El-Rouby
Fig. 5: Photomicrographs of semithin sections in the thyroid gland of rats
from the experimental group (Group II):
a. Shows that the follicles have vacuolated colloid (C). The thyrocytes
have vacuolated cytoplasm (V) and indented darkly stained
pyknotic nuclei (arrow).
Toluidine blue X 1000.
b. Shows that the destroyed follicles reveal shedded cells into
their lumina (encircled).The nuclei of the shedded cells appear
irregular and darkly stained. The thyrocytes reveal vacuolated
cytoplasm (V).
Toluidine blue X 1000.
Fig. 6: A Photomicrograph of semithin section in the thyroid gland of a
rat from the experimental group (group II). It shows a distorted follicle
with shedded cells (S) into its vacuolated colloid (C). These shedded cells
have irregular indented darkly stained nuclei. The colloid (C) reveals
variable density of staining. The thyrocytes show vacuolated cytoplasm
(V). The surrounding stroma is very vascular. Mast cell (M) is seen close
to interstitial blood vessel.
Toluidine blue X 1000.
Fig. 7: A Photomicrograph of TEM section showing two thyrocytes of
two adjacent follicles of a rat from the experimental group, the upper
left thyrocyte is of a damaged follicle showing dilated rER (d rER). Its
nucleus is hetrochromatic and indented (arrows). The lower thyrocyte is
related to more or less normal follicle with normal cisternae of rER but
with apparently more darkly stained lysosome (L). Note the projecting
microvilli (Mv).
Original mag X 6000.
218
Fig. 8: A Photomicrograph of TEM section showing two thyrocytes
of a thyroid follicle of a rat from the experimental group. It shows
dilated cisternae of rER (d rER) with amorphous material (*). The
nuclei (N) appear indented with dilated peri-nuclear space (arrows).
The nuclei have prominent nucleoli (Nu) and hetrochromatin (h).
Original mag X 6000.
Fig. 9: A Photomicrograph of TEM section of a thyrocyte from a rat
of the experimental group reveals apical microvilli (Mv) projecting into
the lumen that has colloid (C). Many darkly stained lysosomes (L) are
seen. The cytoplasm shows prominent Golgi complex (G) and cisternae
of rER.
Original mag X 8000.
Fig. 10: A Photomicrograph of TEM section of the thyroid gland from a
rat of the experimental group (group II). It shows two follicular cells with
flattened nuclei. The nuclei have hetrochromatin. The cytoplasm shows
darkly stained lysosomes (L). The colloid (C) is apparently homogenous.
Original mag X 2000.
A Histological Study on the Effect of Diclofenac Sodium (Declophen) Administration on Thyroid Follicular Cells of Albino Rats
Fig. 14: A photomicrograph of TEM section of the thyroid gland
from a rat of the recovery group (Group III) showing two thyrocytes
with more or less normal rER. Their nuclei appear euchromatic but
with some irregularity. The cytoplasm reveals many vacuoles (V) and
mitochondria (M). Many electron dense lysosomes (L) are observed.
Original mag X 6000.
Fig. 11: A photomicrograph of thyroid gland of a rat from the recovery
group (Group III). It reveals some recovered follicles. These follicles
show vacuolated colloid (V). Other follicles appear distended with
colloid (C) and lined with flat epithelial cells. Note that there are still
some distorted follicles (D). A forth group of involuted (I) follicles are
seen.
H&E X 200.
Fig. 12: A photomicrograph of semithin section in the thyroid gland of
a rat from the recovery group (Group III). It shows that the follicles are
filled with homogenous colloid (C) and lined with cuboidal cells that
have vesicular rounded nuclei (curved arrows) and some low cubical
cells with flattened nuclei (straight arrows). Some mast cells (m) are seen
in the stroma.
Toluidine blue X 1000.
Fig. 15: A photomicrograph of TEM section of a thyrocyte from a rat
of the recovery group (Group III) showing two nuclei with prominent
nucleoli. The cytoplasm shows vacuoles (V) and electron dense
lysosomes (L).
Original mag X 6000.
DISCUSSION
There is a list of medications that alter thyroid
function tests. This list include furosemide, NSAIDs
(salicylate, diclofenac sodium), heparin, amiodarone and
iodinated contrast media12. As NSAIDs are commonly
used drugs so thyroid dysfunction is a potentially
forgotten complication of NSAIDs therapy5.
Fig. 13: A photomicrograph of TEM section of the thyroid gland of a
rat from the recovery group (group III). It shows a follicular cell with
more or less euchromatic nucleus (N) and prominent nucleolus (nu). The
cytoplasm reveals more or less normal cisternae of rER, many vacuoles
(V) and some electron dense lysosomes (L) The lumen shows colloid (C)
.
Original mag X 6000.
219
In the current study, declophen was administered to
rats of group II daily for one week to evaluate its effect
on the structure and function of the thyroid gland. Group
III received declophen for the same duration as group II
and this was followed by another week without treatment
for recovery to asses the reversibility of these changes.
Death of rats that occurred during the experiment was
most probably due to liver and/or kidney affection9,13.
The present study reinforced the previous recorded
results8,14-16 which mentioned that several NSAIDs
Nadia M. El-Rouby
including diclofenac sodium could lower serum T3 while
T4 and TSH were not affected. This might be because
NSAIDs block peripheral (tissue) conversion of T4 to
T3 and produce a low concentration of T3 as most of T3
is produced from peripheral conversion of T4 to T3 by
enzyme 5- deiodinase. The levels of T4 and TSH were not
affected. This was most probably because T4 is strongly
bound to plasma proteins and its biological half life is
long (6-7 days) so subsequently TSH remained without
change17. Another cause for insignificant change in T4
level could be because the thyroid gland is a unique
gland that stores its hormones extracellular in an amount
sufficient for weeks18. NSAIDs including declophen
affected the tests of the thyroid gland through alteration
in the synthesis, transport, and metabolism of thyroid
hormones1,3,5,16. This was proved by both light and
electron microscopic findings. At the light microscopic
level; some follicles showed degenerative changes in the
form of shedded degenerated thyrocytes. The thyrocytes
of more affected follicles had vacuolated cytoplasm and
small, irregular dark pyknotic nuclei. At the level of TEM;
the nuclei of thyrocytes were indented, heterochromatic
with widening of the peri-nuclear space. This might be
attributed to its connection to dilated cisternae of rER. The
rER cisternae were dilated and had intra-luminal electron
lucid material of stored unprocessed protein. Also there
was apparent increase in dark stained lysosomes.
Some follicles revealed variable staining density of
colloid. This might indicate variable follicular activities.
This finding was supported by Wollman et al.23 who
mentioned that density of staining of the accumulated
colloid varied from follicle to follicle and correlated the
staining reaction of the colloid with the activity of the
follicle. Other follicles had vacuolated colloid which
might be due to defect in pinocytosis of follicular cell.
Thyrocytes of the more affected follicles revealed
pyknotic nuclei and vacuolated cytoplasm. This could
explain the lowered level of thyroid hormone (T3)14.
Ultrastructurally, the follicular cells of the damaged
follicles revealed widening of the peri-nuclear space,
irregularity and indentation of the nuclei with chromatin
condensation, dilatation of rER with intra-luminal electron
lucid material, apparently increased darkly stained
lysosomes and prominent Golgi complex. Widening of
the peri-nuclear space might be due to shrinkage of the
nuclei due to the toxic effect of declophen. Also, this might
be attributed to its connection to dilated cisternae of rER.
The pyknosis of the nuclei and condensation of chromatin
observed in thyrocytes of declophen treated rats have been
also observed in other tissues as rat liver cells exposed
to NSAIDs24. These changes could be degenerative
changes. Declophen might induce these degenerative
changes either directly or indirectly. This finding was
in agreement with Boelsterli25 and Moorthy et al.26 who
observed that declophen caused injury of liver cells and
they referred this to immune mediated response. Many
darkly stained lysosomes were seen in the cytoplasm of
thyrocytes of rats administered declophen. The apparent
increased number of lysosomes could be explained by
the accumulation of highly iodinated thyroglobulin that
was resistant to proteolysis27. Marked dilatation of rER
was an evidence of disturbed protein synthesis. This
dilatation could be due to retention of aberrant protein
within their cisternae. Therefore, this might be a form of
rER storage disease. This protein could not be processed,
folded and transported to appropriate sites. Disruption in
protein production might prevent synthesis of apoptosis
inhibitors and/or loss of essential proteins involved in
cellular homeostasis leading to cellular degeneration28,29.
The dilated rER might be the cause of nuclear indentation
and irregularity as the dilated rER compressed the nucleus
causing its indentation and irregularity.
Light microscopic examination of thyroid glands
of rats of the experimental group (Group II) revealed
marked distortion of follicular structure. Some follicles
appeared distended and lined by flat follicular cells
due to increased their colloid content that had minimal
peripheral vacuolations denoting hypoactivity of these
follicles. Other follicles appeared small in size with
apparently little colloid. These follicles might be newly
formed, collapsed or involuted follicles. A third group of
follicles had shedded epithelial cells. These changes could
be attributed to cellular distension with accumulated
colloid which resulted in cellular disruption and collapse
with subsequent collapse of the follicles. The degenerated
cells had dark stained nuclei and vacuolated cytoplasm.
These findings were recorded in cases of thyroiditis19.
Many mast cells were observed frequently in the interfollicular connective tissue beside blood vessels. This
finding might be due to an allergic reaction occurred in
the thyroid gland as these mast cells secrete cytokines
and chemical mediators responsible for allergic reaction.
This allergic reaction might be because NSAIDs altered
plasma proteins. These NSAIDs- altered proteins could
become antigenic. By time these antigens would become
immunogens20. Former studies reported that mast cells
participate in the process of thyroid hormone secretion
and in thyroid gland activity. Mast cells might mediate
the action of TSH on follicular cells. TSH has been
shown to promote the release of serotonin from thyroid
mast cells. Serotonin activates thyroid follicular cells
by enhancing them to extend pseudopodia and engulf
thyroglobulin from the follicular lumen21,22.
Using light microscopy, the thyroid gland of the
recovery group revealed that the gland needed more
time to return back to normal structure. The thyroid
follicles revealed patchy recovery. Some follicles were
still large in diameter and distended with colloid with
minimal peripheral vacuolations which indicated hypoactivity. Other follicles were smaller in diameter, lined
with cuboidal cells mainly and their colloid had some
peripheral vacuolations i.e. these follicles regain their
activity.
Ultrastructurally, the thyrocytes of that group
220
A Histological Study on the Effect of Diclofenac Sodium (Declophen) Administration on Thyroid Follicular Cells of Albino Rats
appeared more or less normal with slightly indented
euchromatic nuclei and prominent nucleoli. The
cytoplasmic organelles were more or less normal in
appearance except for the presence of apparently more
darkly stained lysosomes. Some thyrocytes revealed
variable sized vacuoles in their cytoplasm. This indicated
that rat thyroid needed more time to recover completely.
4.
5.
6.
Morphometeric study of thyroid follicles of rats of
the experimental group (Group II) revealed statistically
significant increase in the mean follicular diameter
and statistically significant decrease in the mean
epithelial height in comparison with the control group
(Group I) which denoted hypoactivity of the gland.
This was proved histologicaly as many follicles were
markedly distended with colloid and lined mainly with
flat cells. The colloid was homogenous acidophilic and
showed minimal peripheral vacuolations. The less active
follicles of the thyroid gland were distended by stored
colloid and the lining cells appear flattened against the
follicular basement membrane17,30.
7.
8.
9.
One week after declophen withdrawal (recovery
group; group III), the morphometeric analysis of the
thyroid follicles showed insignificant increase in the
mean follicular diameter of thyroid follicles and also
insignificant decrease in the epithelial height of the lining
follicular cells in comparison to the control group. This
could be attributed to starting reactivity of the thyroid
gland as a result of recovery from the toxic effect of
declophen and elimination of harmful waste products
from the body.
10.
11.
12.
13.
CONCLUSION
14.
Declophen, a cytotoxic drug, induced variable
structural and functional alterations in the thyroid
follicular cells of adult male albino rats. These alterations
were incompletely recovered. So, it is advisable to avoid
using declophen as first pain killer and if it is used, thyroid
function tests’ results should be interpreted cautiously
in patients on declophen therapy. The drug should be
discontinued prior to evaluation of thyroid function.
15.
16.
17. Ganong WF. (2005): Review of medical physiology. 22nd ed.
McGraw-Hill Medical.
18. Wheater PR, Burkitt HG and Victor G. Daniels VG. (1987):
Functional histology: Text and colour atlas.2nd ed. Churchill
Livingstone.
19. Smyrk TC, Goellner JR, Brennan MD and Carney JA. (1987):
Pathology of the thyroid in amiodarone-associated thyrotoxicosis.
Am. J. Surg. Pathol. Mar; 11(3):197-204.
20. Boelsterli UA, Zimmerman HJ and Kretz Rommel A. (1995):
Idiosyncratic liver toxicity of nonsteroidal antiinflammatory
drugs: Molecular mechanisms and pathology. Crit. Rev.
Toxicol.; 25(3):207-235.
21. Melander A and Sundler F. (1972): Significance of thyroid
mast cells in thyroid hormone secretion. Endocrinology
Mar; 90(3):802-807.
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‫دراﺳﺔ ﺗﺄﺛﻴﺮ ﻋﻘﺎر اﻟﺪﻳﻜﻠﻮﻓﻴﻦ ﻋﻠﻰ اﻟﺨﻼﻳﺎ اﻟﻤﺒﻄﻨﺔ ﻟﺤﻮﻳﺼﻼت اﻟﻐﺪة اﻟﺪرﻗﻴﺔ ﻓﻲ اﻟﻔﺌﺮان‬
‫اﻟﺒﻴﻀﺎء‬
‫ﻧﺎدﻳﺔ ﻣﺤﻤﻮد ﻣﺤﻤﺪ اﻟﺮوﺑﻲ‬
‫ﻗﺴﻢ اﻟﻬﺴﺘﻮﻟﻮﺟﻴﺎ‪ ،‬آﻠﻴﺔ ﻃﺐ اﻟﻘﺼﺮ اﻟﻌﻴﻨﻲ‪ ،‬ﺟﺎﻣﻌﺔ اﻟﻘﺎهﺮة‬
‫ﻣﻠﺨﺺ اﻟﺒﺤﺚ‬
‫ﻳﻌﺪ ﻋﻘﺎر اﻟﺪﻳﻜﻠﻮﻓﻴﻦ ﺣﺪﻳﺜﺎ أﺣﺪ اﻷدوﻳﺔ اﻟﺘﻲ ﻗﺪ ﺗﺆﺛﺮ ﻋﻠﻰ اﻟﻐﺪة اﻟﺪرﻗﻴﺔ وﻗﺪ أﺟﺮﻳﺖ هﺬﻩ اﻟﺪراﺳﺔ ﻋﻠﻰ اﻟﻔﺌﺮان ﻟﻤﻌﺮﻓﺔ‬
‫ﻣﺪى ﺗﺄﺛﻴﺮ هﺬا اﻟﻌﻘﺎر ﻋﻠﻰ اﻟﻐﺪة اﻟﺪرﻗﻴﺔ ﻣﻦ اﻟﻨﺎﺣﻴﺘﻴﻦ اﻟﺘﺮآﻴﺒﻴﺔ واﻟﻮﻇﻴﻔﻴﺔ وﻣﺎ إذا آﺎﻧﺖ هﺬﻩ اﻟﺘﻐﻴﺮات ﻣﺆﻗﺘﺔ أم ﻻ‪.‬‬
‫أﺟﺮﻳﺖ هﺬﻩ اﻟﺪراﺳﺔ ﻋﻠﻰ ﺛﻼﺛﻴﻦ ﻓﺄرا‪ ،‬ﻗﺴﻤﺖ إﻟﻰ ﺛﻼث ﻣﺠﻤﻮﻋﺎت ﻣﺘﺴﺎوﻳﺔ‪ ،‬آﺎﻟﺘﺎﻟﻲ‪:‬‬
‫اﻟﻤﺠﻤﻮﻋﺔ اﻷوﻟﻰ )اﻟﻤﺠﻤﻮﻋﺔ اﻟﻀﺎﺑﻄﺔ( واﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻧﻴﺔ )اﻟﻤﺠﻤﻮﻋﺔ اﻟﻤﻌﺎﻟﺠﺔ( ﻗﺪ ﺗﻢ ﺣﻘﻨﻬﺎ ﺑﺎﻟﻌﻘﺎر ﺑﺠﺮﻋﺔ ‪8‬‬
‫ﻣﺠﻢ‪ /‬آﺠﻢ ﻣﻦ وزن اﻟﺠﺴﻢ ﻳﻮﻣﻴﺎ وذﻟﻚ ﻟﻤﺪة أﺳﺒﻮع‪ .‬أﻣﺎ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻟﺜﺔ )اﻟﻤﺠﻤﻮﻋﺔ اﻟﻤﺘﻤﺎﺛﻠﺔ ﻟﻠﺸﻔﺎء( ﻗﺪ ﺗﻢ ﺣﻘﻨﻬﺎ‬
‫ﺑﺎﻟﻌﻘﺎر ﻣﺜﻞ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻧﻴﺔ ﺛﻢ ﺗﺮآﺖ ﻟﻤﺪة أﺳﺒﻮع أﺧﺮ ﺑﺪون ﻋﻼج‪.‬‬
‫ﺗﻢ ﺗﺸﺮﻳﺢ اﻟﻔﺌﺮان ﻓﻲ اﻷوﻗﺎت اﻟﻤﺤﺪدة ﻟﺬﻟﻚ آﻤﺎ ﺗﻢ أﺧﺬ ﻋﻴﻨﺎت دم ﻣﻨﻬﺎ ﻟﻘﻴﺎس ﻣﺴﺘﻮى هﺮﻣﻮﻧﺎت اﻟﻐﺪة اﻟﺪرﻗﻴﺔ وآﺬﻟﻚ‬
‫اﻟﻬﺮﻣﻮن اﻟﻤﺤﻔﺰ ﻟﻬﺎ‪ .‬و ﺗﻢ أﺧﺬ اﻟﻐﺪة اﻟﺪرﻗﻴﺔ ﻣﻦ اﻟﻔﺌﺮان وﺗﺤﻀﻴﺮهﺎ ﻟﻠﻔﺤﺺ اﻟﻤﻴﻜﺮوﺳﻜﻮﺑﻲ ‪ ،‬آﻤﺎ ﺗﻢ ﻗﻴﺎس ﻣﺘﻮﺳﻂ‬
‫ﻗﻄﺮ ﺣﻮﻳﺼﻼت اﻟﻐﺪة اﻟﺪرﻗﻴﺔ وآﺬﻟﻚ ﻣﺘﻮﺳﻂ ارﺗﻔﺎع اﻟﻨﺴﻴﺞ اﻟﻄﻼﺋﻲ اﻟﻤﺒﻄﻦ ﻟﻬﺎ وﺣﻠﻠﺖ اﻟﻨﺘﺎﺋﺞ اﺣﺼﺎﺋﻴﺎ ‪،‬‬
‫أوﺿﺤﺖ اﻟﺪراﺳﺔ أن اﻟﺪﻳﻜﻠﻮﻓﻴﻦ ﻗﺪ ﺗﺴﺒﺐ ﻓﻲ ﻧﻘﺺ ذو دﻻﻟﺔ اﺣﺼﺎﺋﻴﺔ ﻓﻲ هﺮﻣﻮن اﻟﻐﺪة اﻟﺪرﻗﻴﺔ اﻟﺜﻼﺛﻲ )‪ (T3‬وذﻟﻚ‬
‫ﻓﻲ اﻟﻤﺠﻤﻮﻋﺔ اﻟﻤﻌﺎﻟﺠﺔ ﺑﺎﻟﻤﻘﺎرﻧﺔ ﻣﻊ اﻟﻤﺠﻤﻮﻋﺔ اﻟﻀﺎﺑﻄﺔ‪.‬‬
‫آﻤﺎ أﻇﻬﺮ اﻟﻤﻴﻜﺮﺳﻜﻮب اﻟﻀﻮﺋﻲ ﺑﻌﺾ اﻟﺘﻐﻴﺮات ﻓﻲ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻧﻴﺔ وذﻟﻚ ﻓﻲ ﺻﻮرة اﻧﺘﻔﺎخ ﻓﻲ ﺑﻌﺾ اﻟﺤﻮﻳﺼﻼت‬
‫اﻟﺘﻲ آﺎن ﻧﺴﻴﺠﻬﺎ اﻟﻄﻼﺋﻲ ﻣﺴﻄﺤﺎ وﻣﻤﺘﻠﺌﺔ ﺑﺎﻟﻬﺮﻣﻮن اﻟﻐﺮوي واﻟﺒﻌﺾ اﺣﺘﻮى ﻋﻠﻰ ﺧﻼﻳﺎ ﻣﺘﺴﺎﻗﻄﺔ وأﻇﻬﺮت‬
‫اﻟﺤﻮﻳﺼﻼت اﻷآﺜﺮ ﺗﺄﺛﺮًا ﺗﻐﻴﺮات ﻓﻲ ﺧﻼﻳﺎهﺎ اﻟﻤﺒﻄﻨﺔ وذﻟﻚ ﻓﻲ ﺻﻮرة ﻓﺮاﻏﺎت ﺳﻴﺘﻮﺑﻼزﻣﻴﺔ وﺗﺪﻣﻴﺮ ﺑﺄﻧﻮﻳﺘﻬﺎ وﻟﻢ‬
‫ﻳﻈﻬﺮ اﻟﻬﺮﻣﻮن اﻟﻐﺮوي ﻓﻲ اﻟﺒﻌﺾ ﻣﻦ اﻟﺤﻮﻳﺼﻼت اﻻ ﺑﻜﻤﻴﺎت ﻗﻠﻴﻠﺔ‪.‬‬
‫وأوﺿﺢ اﻟﻤﻴﻜﺮﺳﻜﻮب اﻹﻟﻜﺘﺮوﻧﻲ اﻧﺘﻔﺎخ ﻓﻲ اﻟﺸﺒﻜﺔ اﻷﻧﺪوﺑﻼزﻣﻴﺔ اﻟﺨﺸﻨﺔ ﻣﻊ وﺟﻮد ﻣﻮاد ﻣﺨﺰﻧﺔ ﺑﺘﺠﺎوﻳﻔﻬﺎ وزﻳﺎدة‬
‫واﺿﺤﺔ ﻓﻲ اﻟﻠﻴﺴﻮزوﻣﺎت ﻣﻊ ﺗﻌﺮﻳﺞ ﻓﻲ ﺷﻜﻞ اﻟﻨﻮاة وﺑﻘﻴﺖ ﺑﻌﺾ هﺬﻩ اﻟﺘﻐﻴﺮات ﻣﻮﺟﻮدة ﻓﻲ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻟﺜﺔ ‪.‬‬
‫وأﻇﻬﺮ اﻟﺘﺤﻠﻴﻞ اﻻﺣﺼﺎﺋﻲ ﻟﻠﻨﺘﺎﺋﺞ زﻳﺎدة ذو دﻻﻟﺔ اﺣﺼﺎﺋﻴﺔ ﻓﻲ ﻣﺘﻮﺳﻂ ﻗﻄﺮ اﻟﺤﻮﻳﺼﻼت ﻓﻲ اﻟﻤﺠﻤﻮﻋﺔ اﻟﺜﺎﻧﻴﺔ ﻣﻊ‬
‫ﻧﻘﺺ ﻓﻲ ﻣﺘﻮﺳﻂ ارﺗﻔﺎع اﻟﻨﺴﻴﺞ اﻟﻄﻼﺋﻲ اﻟﻤﺒﻄﻦ ﻟﻬﺎ ﺑﺎﻟﻤﻘﺎرﻧﺔ ﺑﺎﻟﻤﺠﻤﻮﻋﺔ اﻟﻀﺎﺑﻄﺔ ‪.‬‬
‫وﻳﻤﻜﻦ اﻻﺳﺘﻔﺎدة ﻣﻦ هﺬﻩ اﻟﺪراﺳﺔ ﻓﻲ أﺧﺬ اﻟﺤﻴﻄﺔ ﻋﻨﺪ اﺟﺮاء ﺗﺤﻠﻴﻞ اﻟﻐﺪة اﻟﺪرﻗﻴﺔ ﻓﻲ اﻷﺷﺨﺎص اﻟﺨﺎﺿﻌﻴﻦ ﻟﻠﻌﻼج‬
‫ﺑﻌﻘﺎر اﻟﺪﻳﻜﻠﻮﻓﻴﻦ‪.‬‬
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