Download Light and Electron Microscopic Study on the Effect of

Egypt. J. Histol. Vol. 32, No. 1, June, 2009: 101 - 108
(ISSN: 1110 - 0559)
Original Article
Light and Electron Microscopic Study on the Effect of Lithium Carbonate
on the Thyroid Gland of Adult Male Albino Rat
Nafisa A. El-Bakary and Gehan M. Soliman
Histology Department, Faculty of Medicine, Tanta University
ABSTRACT
Introduction: Lithium carbonate is the treatment of choice for acute manic episodes. It is often referred to as an antimanic drug as it prevents mood swings in patients with manic- depressive disorder. Thyroid disturbances during lithium
treatment had been reported.
Aim of the Work: This research was performed to study the effect of lithium carbonate on the thyroid gland of albino
rat and the possibility of recovery after drug withdrawal.
Materials and Methods: Thirty adult male albino rats were divided into three equal groups; Group I (Control Group),
group II received lithium carbonate at a daily dose of 14.4 mg for each rat for 6 weeks orally and group III received
the same dose of lithium carbonate as group II and then left untreated for another 6 weeks to study the possibility of
recovery after the drug withdrawal. The specimens were prepared for light and electron microscopic examination.
Results: Sections of lithium carbonate treated rats showed enlarged irregular shaped thyroid follicles with papillary
infoldings projecting into the follicular lumena. Detached and desquamated follicular cells were seen in the follicular
colloid. The follicular cells showed apparent hyperplasia and bizarre-appearing nuclei. The interstitial tissue showed
cellular infiltration, presence of deeply eosinophilic large cells and fibrosis in some specimens. Ultrastructurally, there
was cellular debris in the follicular lumena. Some follicles showed dark follicular cells containing electron dense
cytoplasm and indistinct organelles. The above structural changes were much less pronounced in group III (Recovery
Group).
Conclusion: Lithium carbonate induced histological changes in the thyroid gland of albino rat and most of these
changes were seen to be improved after withdrawal of the drug. So, the use of this drug should be justified in clinical
situation under direct medical supervision.
Key Words: Lithium carbonate, thyroid gland,
Corresponding Author: Nafisa A. Elbakary
histopathology.
Tel.: 0109171524
E-mail: [email protected]
INTRODUCTION
excreted unchanged in the urine4. It has been reported
that lithium induced gastrointestinal, neuromuscular and
endocrine adverse effects in patients taking it5.
Lithium salts are used as mood stabilizing drugs.
They have a role in the treatment of depression and
mania in both acute and long term conditions1. Lithium
carbonate is the treatment of choice for acute manic
episodes. It is often referred to as an anti-manic drug. It
prevents mood swings in patients with manic-depressive
disorder2. International multi center studies yield strong
evidence that mortality and suicide rates could be
lowered by long- term lithium treatment3.
The thyroid gland is a highly vascular organ. The
pathway of thyroid hormones synthesis, transport of
secretion in the circulation and their metabolism offer
numerous targets for drug interaction6.
Thyroid disturbances during lithium treatment had
been reported6. However, there have been few reports
describing the histological alterations of the thyroid
gland during lithium treatment. This research was
carried out to study the effect of lithium carbonate on
the structure of the thyroid gland of albino rat and the
possibility of recovery after drug withdrawal.
Lithium carbonate is virtually totally absorbed
from the gastrointestinal tract within 18 hours of oral
administration, with peak blood levels achieved in 2 to
4 hours. Its tissue uptake is not uniform, with levels in
the brain, thyroid and saliva exceeding plasma levels.
Lithium is not bound to proteins or metabolized but
9 (1134-2009)
101
Light and Electron Microscopic Study on the Effect of Lithium Carbonate on the Thyroid Gland
MATERIALS AND METHODS
of thyroid follicles of variable sizes and their lumena
contained colloid. The follicles were lined by simple
cuboidal epithelium (follicular epithelium) (Fig. 1).
Mallory’s trichrome stained sections showed little
amount of connective tissue in interstitial tissue between
the thyroid follicles (Fig. 2).
This research was carried out on 30 adult male albino
rats ranging in weight from 150-200 grams. All animals
were maintained on a standard diet and water and kept
under suitable conditions. They were divided into three
groups:
Group II: Examination of specimens obtained from
lithium-treated animals showed morphological changes
in the thyroid glands. Many specimens showed irregular
shaped follicles with papillary infoldings projecting
into the follicular lumena. Extra follicular colloid was
evident in the inter follicular spaces (Fig. 3). Some
follicles were lined by simple squamous, others were
lined by simple cuboidal or tall columnar epithelium
(Figs. 4&5). Bizarre-appearing nuclei in the form of
pleomorphism and hyperchromasia were also observed
in the lining epithelium of some follicles and scalloped
edges of the colloid of some follicles were detected.
(Fig. 5). Detached and desquamated follicular cells were
present in the follicular colloid. Some follicles were filled
with aggregates of these desquamated follicular cells
(Figs. 6&7). Pinched apical parts of follicular cells
associated with apical vacuolation were observed
(Figs. 8&9).Cellular infiltration in the interstitial tissue
was detected (Fig. 9). Deeply eosinophilic large cells
were present in the interstitial tissue in many specimens
(Fig. 10). Mallory’s trichrome stained sections showed
interstitial fibrosis in some specimens (Fig. 11).
Group I (Control Group): Included 10 rats which
were further subdivided into 2 subgroups. Subgroup A
(negative control animals) included 5 animals being kept
without any treatment. Subgroup B (positive control)
included 5 animals each received distilled water by
the same route, dose and duration as lithium carbonate
treated animals.
Group II: Included 10 rats, each received lithium
carbonate (Prianil CR) at a daily dose of 14.4 mg7,8 for 6
weeks. This drug is manufactured by EL-Nile Company.
It is present in the form of tablets of 400 mg. The drug
was dissolved in distilled water and the calculated dose
was given per mouth using feeding tubes.
Group III: Included 10 rats, each received the same
dose of lithium carbonate as group II for the same period
and then left untreated for another 6 weeks to study the
possibility of recovery after the drug withdrawal.
At the expected time, the animals were anaesthetized
with diethyl ether and perfused intracardially with 4%
paraformaldehyde in 0.1 M phosphate buffered solution
(PH 7.2) containing 1 % glutaraldehyde solution. The
thyroid glands were dissected out and some specimens
were processed for light microscopy and stained with
haematoxylin and eosin and Malloryʼs trichrome9.
Group III: Examination of specimens obtained
from group III (Recovery Group) showed regression of
the lesions in many specimens. However, some thyroid
follicles still showed papillary infoldings and irregular
outlines (Fig. 12). Mallory’s trichrome stained sections
showed interstitial fibrosis in some sections (Fig. 13).
Parallel tissue specimens were processed for
electron microscopy by immediate immersion for
one hour in a glutaraldehyde fixative based on 0.1 M
phosphate buffer (pH 7.2). They were then washed three
times (5 minutes each) with phosphate buffer followed
by postfixation in 1 % phosphate buffered osmium
tetroxide for thirty minutes at room temperature. After
dehydration in ascending grades of ethyl alcohol, the
specimens were embedded in Epon–araldite mixture.
Ultrathin sections were cut with ultramicrotome using
a diamond knife and stained with uranyl acetate and
lead citrate10 to be examined by the transmission
electron microscope at Faculty of Science, Ain-Shams
University.
Electron microscopic results:
Group I: Electron microscopic examination of
specimens obtained from the control animals showed a
part of thyroid follicle composed of cuboidal follicular
cells with rounded nuclei. The apical surfaces of
these cells showed small microvilli protruded into the
follicular lumen that contained colloid (Fig. 14).
Group II: Examination of specimens obtained from
lithium-treated animals revealed cellular debris in the
follicular lumen (Figs. 15&16). Some follicles showed
dark follicular cells which contained electron dense
cytoplasm and indistinct organelles (Fig. 17).
RESULTS
Group III: Examination of specimens obtained from
animals of recovery group showed that thyroid follicles
appeared more or less as the control group but some
follicular cells showed dense bodies in their cytoplasm
(Fig. 18).
Light microscopic results:
Group I: Light microscopic examination of
specimens from all control animals showed the same
histological structures. The thyroid gland consisted
102
Nafisa A. El-Bakary and Gehan M. Soliman
Fig. 1: A photomicrograph of a section of thyroid gland of control group
showing thyroid follicles lined by simple cuboidal epithelium (→) and
containing colloid.
H&E., Mic. Mag. X 400.
Fig. 2: A photomicrograph of a section of thyroid gland of control
group showing little amount of connective tissue in interstitial
tissue (→).
Mallory's trichrome, Mic. Mag. X 400.
Fig. 4: A photomicrograph of a section of thyroid gland of lithium treated
rats showing irregular shaped follicles, some of them are lined by simple
squamous epithelium (→) and others are lined by simple columnar
epithelium (►). Notic the presence of papillary projections in some
follicles.
H&E., Mic. Mag. X 200.
Fig. 5: A photomicrograph of a section of thyroid gland of lithium treated
rats showing parts from thyroid follicles one is lined by tall columnar
epithelium (►) and the other is lined by cuboidal epithelium with
pleomorphic nuclei (→). Notice the scalloped appearance of the colloid
edges (*).
H&E., Mic. Mag. X 1000.
Fig. 3: A photomicrograph of a section of thyroid gland of
lithium treated rats (Group II) showing irregular shaped follicles
with papillary infoldings projecting into the lumena (→).
Extrafollicular colloid (*) is evident in the interfollicular spaces.
H&E., Mic. Mag. X 100.
103
Fig. 6: A photomicrograph of a section of thyroid gland of lithium treated
rats showing detached (→) and desquamated (►) follicular cells in the
colloid of the follicles.
H&E., Mic. Mag. X 200.
Light and Electron Microscopic Study on the Effect of Lithium Carbonate on the Thyroid Gland
Fig. 7: Higher magnification of the previous figure showing the detached
follicular cells in the colloid (→).
H&E., Mic. Mag. X 400.
Fig. 8: A photomicrograph of a section of thyroid gland of lithium treated
rats showing pinched apical parts of follicular cells (→) associated with
apical vacuolation (►).
H&E., Mic. Mag. X 1000.
Fig. 9: A photomicrograph of a section of thyroid gland of lithium
treated rats showing cellular infiltration in the interstitial tissue (*) and
cytoplasmic vacuolation of follicular cells (→).
H&E., Mic. Mag. X 1000.
Fig. 10: A photomicrograph of a section of thyroid gland of lithium
treated rats showing large eosinophilic cells in the interstitial tissue
(→). Notice that the follicular cells have hyperchromatic nuclei.
H&E., Mic. Mag X 1000.
Fig. 11: A photomicrograph of a section of thyroid gland of lithium
treated rats showing fibrosis of interstitial tissue (*).
Mallory’s trichrome, Mic. Mag. X 400.
Fig. 12: A photomicrograph of a section of thyroid gland of group III
(Recovery Group) showing some thyroid follicles appeared similar to
those of the control, while others still containing papillary infoldings (*).
H&E., Mic. Mag. X 200.
104
Nafisa A. El-Bakary and Gehan M. Soliman
Fig. 13: A photomicrograph of a section of thyroid gland of recovery
group showing interstitial fibrosis (*).
Mallory’s trichrome, Mic. Mag. X 400.
Fig. 14: An electron micrograph of a section of thyroid gland of
control group showing a part of thyroid follicle lined by cuboidal cells
with rounded nuclei. The apical surfaces of these cells showed small
microvilli (→) protruded into the follicular lumen (L) that contains
colloid.
Mic. Mag. X 3000.
Fig. 15: An electron micrograph of a section of thyroid gland
of lithium treated rats showing cellular debris in the follicular
lumen (→).
Mic. Mag. X 3000.
105
Fig. 16: Higher magnification of the previous figure showing cellular
debris in the follicular lumen (→).
Mic. Mag. X 6000.
Fig. 17: An electron micrograph of a section of thyroid gland of lithium
treated rats showing two adjacent follicular cells, one of them (F) showed
dark cytoplasm and indistinct organelles as compared with the other
cell (F1). Notice, microvilli (→) protruded into the lumen (L).
Mic. Mag. X 6000.
Fig. 18: An electron micrograph of a section of thyroid gland of recovery
group showing the follicular cells appear more or less similar to control.
Some cells contain dense bodies (→).
Mic. Mag. X 3000.
Light and Electron Microscopic Study on the Effect of Lithium Carbonate on the Thyroid Gland
DISCUSSION
of follicular disruption in lithium treated patients
resembled those found in patients with silent thyroiditis
which were considered to represent a destructive phase
of chronic thyroiditis18. Such follicular destruction
had
also been reported in amiodarone induced
hyperthyroidism19.
This study showed that lithium treatment induced
histological and ultrastructural changes in the thyroid
gland of albino rats. Thyroid disturbances during lithium
treatment had been reported. These commonly presented
as goiter with or without hypothyroidism. Also some
scientists stated that there were cases of hyperthyroidism
among patients treated with lithium11,12. In addition,
lithium associated thyroiditis and lithium associated
autoimmune thyroiditis had been recorded6,13.
In this study deeply eosinophilic cells were observed
in the interstitial tissue in many specimens. Some
investigators described deeply eosinophilic cells with
granular cytoplasm called Hurthle cells in Hashimoto
thyroiditis20. Cellular infiltration and interstitial fibrosis
were observed in many specimens of this study. This
appearance is consistent with that found in autoimmune
thyroiditis20. Furthermore, some studies had reported
a higher incidence of thyroid antibodies in patients
treated with lithium compared with those not taking
lithium21.
In this research the thyroid glands of lithium treated
animals were composed of large distended follicles with
abundant colloid. Some of these follicles were lined
with simple squamous epithelium, but many follicles
were lined with tall columnar epithelium which showed
hyperplasia and papillary infoldings projecting into the
thyroid lumena.
As regards to the ultrastructural findings, cellular
debris in the follicular lumena, degenerated dark
follicular cells in some follicles with indistinct organelles
and detached apical parts of the cytoplasm of some
follicular cells were observed in this study. These
ultrastructural findings confirmed the light microscopic
results and indicated that lithium could induce follicular
cell damage and follicular disruption.
The same results were reported by other
investigators7. In this regard, some authors stated that
lithium associated changes of the thyroid glands were
similar to the changes seen in radiation –treated glands
including the presence of nodular hyperplasia associated
with increased cellularity and fibrosis14. The bizarreappearing nuclei of some follicular cells seen in this
study were also recorded by other investigators who
reported that in chronic lithium therapy the nuclei of
follicular cells showed nucleomegaly, pleomorphism
and hyperchromasia14. In this research, scalloping of
the colloid edges was detected in many follicles in
lithium treated animals. It was stated that the enlarged
epithelial cells projecting into the lumena of the
follicles actively resorb the colloid in the centers of the
follicles, resulting in the scalloped appearance of the
edges of the colloid15.
In this investigation, the changes observed in
the thyroid gland described above were no longer
encountered after the drug withdrawal. However, few
follicles still showed few papillary infoldings. Also;
stromal fibrosis in some specimens was also observed.
Ultrastructurally, the thyroid follicles appeared more
or less similar to control except the presence of
dense bodies in some follicular cells. In this regard,
it was reported that some patients became euthyroid
spontaneously with cessation of lithium treatment7.
The mechanism of the effect of lithium on the
thyroid gland is not fully understood. However, it had
been reported that lithium is a monovalent cation that
becomes concentrated in the thyroid gland16. It had been
shown that lithium reduces iodine uptake into the gland,
impairs coupling of iodotyrosines and interferes with
the release of hormone from the gland7.
CONCLUSION
From this research, it could be suggested that lithium
carbonate induced severe histological changes in the
thyroid gland of albino rat and most of these changes
were seen to be improved after withdrawal of the drug.
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In this study follicular disruption with release
of colloid in the interfollicular spaces, detachment
and desquamation of follicular cells and presence of
these desquamated cells in the follicular colloid were
common findings. Occasionally some thyroid follicles
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were similarly described by some investigators7 and
others reported that lithium itself may directly injure
thyroid follicular cells and cause thyroid follicular
destruction17. They added that subsequent release of
thyroglobulin increases the serum thyroid hormone
concentration causing hyperthyroidism. On the other
hand, it was stated that the histological features
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R. (2002): Thyroid abnormalities in lithium-treated patients with
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‫دراسة بالميكروسكوب الضوئي و االلكتروني لتأثير كربونات الليثيوم علي الغدة الدرقية في الجرذ األبيض‬
‫البالغ‬
‫نفيسة عبد الرحيم البقرى ‪ ،‬جيھان محمد سليمان‬
‫قسم الھستولوجيا ‪ ،‬كلية الطب‪ ،‬جامعة طنطا‬
‫ملخص البحث‬
‫ان كربونات الليثيوم ھي العالج المثالي لنوبات الجنون الحادة‪ .‬و لذلك يوصف ھذا العقار في كثير من األحيان كعالج لمنع‬
‫التغيرات المزاجية المفاجئة في مرضي الجنون االكتئابي‪ ،‬و لقد وجد انه يسبب اضطرابات بالغدة الدرقية و لذا فان الھدف من‬
‫ھذا البحث ھو دراسة ھستولوجية لتأثير كربونات الليثيوم علي الغدة الدرقية في الجرذ األبيض و دراسة امكانية النقاھة من تأثير‬
‫ھذا العقار بعد ايقاف اعطاؤه‪.‬‬
‫تم تقسيم ثالثون جرذا الي ثالث مجموعات متساوية‪ ،‬المجموعة األولي ضابطة‪ ,‬المجموعة الثانية تلقت ‪ 14.4‬مجم‬
‫كربونات ليثيوم كجرعة لكل جرذ بالفم يوميا لمدة ‪ 6‬أسابيع‪ ,‬أما المجموعة الثالثة فقد تلقت نفس جرعة كربونات الليثيوم بنفس‬
‫الطريقة و لنفس المدة و لكن تم تركھا لمدة ‪ 6‬أسابيع أخري بعد ايقاف الدواء لدراسة امكانية النقاھة من ھذا الدواء بعد سحبه‪.‬‬
‫و تم تجھيز عينات لفحصھا بالميكروسكوب الضوئي و االلكتروني‪.‬‬
‫و قد أوضحت النتائج في المجموعة الثانية ظھور حويصالت الغدة الدرقية كبيرة و غير منتظمة مع وجود حلمات بارزة‬
‫في تجاويف ھذه الحويصالت و وجود خاليا الحويصالت تالفة و منفصلة في السائل الھالمي الحويصلي‪ .‬كما وجد تزايد ظاھري‬
‫في عدد الخاليا الحويصلية مع عشوائية في األنوية ‪ .‬كما لوحظ وجود ارتشاح خلوي و ظھور خاليا كبيرة حمضية الصبغة‬
‫و تليف في النسيج الخاللى فى بعض العينات ‪ ،‬أما دراسة التركيب الدقيق فقد وجدت نفايا خلوية في تجاويف الحويصالت و‬
‫ظھور بعض الخاليا معتمة ذات أنوية داكنه و صغيرة مع عدم وضوح العضيات‪ ،‬و في المجموعة الثالثة فان كل ھذه التغيرات‬
‫لم تكن ملحوظة بدرجة أو بأخري‪.‬‬
‫نستنتج من ھذا البحث أن عقار كربونات الليثيوم يؤدي الي تغيرات جسيمة في تركيب الغدة الدرقية في الجرذ األبيض‬
‫و لقد تحسنت تلك التغيرات بعد سحب ھذا الدواء‪ .‬و لذلك فانه ينصح بعدم استخدام ھذا العقار اال في حاالت مرضية معينة‬
‫و تحت أشراف طبي مباشر‪.‬‬
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