Toxicity of fluoxetine hydrochloride on some selected vital organs of pregnant mice Mus musculus

Consequently, these lesions have a great impact on the heath of both fetus and pregnant mice since the most common lesions were observed in fetuses which causes abortion which affects the health of pregnant mice.

Toxicity of fluoxetine hydrochloride on some selected vital organs of pregnant mice Mus musculus

Baidaa Abdul-Aziz Mohammed1 and Saevan Saad Al-Mahmood2

1Department of Biology, College of Education for Pure Science, 2Department of Pathology and Poultry Diseases, College of Veterinary Medicine, University of Mosul, Mosul, Iraq

baidaamohammed@uomosul.edu.iq,

saevan981@yahoo.com, 0000-0002-9693-0354

Abstract

The current study intends to look at the impact of the fluoxetine hydrochloride on specific tissues lung and pancreas of mature pregnant mice. The two doses used during the study were 45,75mg/kg b.w. from the 7th to the 17th day of pregnancy. Experimental animals received fluoxetine orally at a dosage of 45mg/kg b.w. The findings indicate variable pathologic changes in the lungs. At the dose of 75 mg/kg b.w. hyperplasia of pneumocytes occurred. In the pancreas, there were no detectible lesions at the dose of 45mg/kg b.w. while at the dose of 75 mg/kg b.w. the severity of tissue lesions was seen. In conclusion, antidepressants may stimulate oxidative injury throughout the body's internal organs, particularly if taken at high doses during pregnancy. Consequently, these lesions have a great impact on the heath of both fetus and pregnant mice since the most common lesions were observed in fetus which cause abortion which effect on the health of pregnant mice.

Keywords: Fluoxetine, Mice, Histopathology, Pregnancy, Lung

سمیة الفلوکستین هایدروکلورید على بعض الأعضاء الحیویة المختارة فی الفئران الحوامل Mus musculus

بیداء عبدالعزیز محمد و سیڤان سعد المحمود

فرع علم الأحیاء، کلیة التربیة للعلوم الصرفة، 2فرع الأمراض وأمراض الدواجن، کلیة الطب البیطری، جامعة الموصل، الموصل، العراق

الخلاصة

أخذت الدراسة الحالیة على عاتقها التحری عن تأثیر عقار فلوکستین هایدروکلوراید على کل من الرئة والبنکریاس فی الفئران البالغة الحوامل. استخدمت فی الدراسة الحالیة جرعتین هما: 45، 75 ملغم /کغم من وزن الجسم. وجرعت الفئران الحوامل فمویا بالعقار من الیوم 7 إلى یوم 17 من الحمل. أظهرت النتائج تغیرات نسجیة مرضیة مختلفة فی الرئة عند الجرعة 45 ملغم /کغم من وزن. ازدادت حدة الآفات النسجیة المرضیة عند الجرعة 75 ملغم /کغم من وزن الجسم، فضلا عن ظهور فرط التنسج فی الخلایا الرئویة. لم تظهر أفات نسجیة مرضیة واضحة عند الجرعة 45 ملغم /کغم من وزن الجسم فی البنکریاس، فی حین عند الجرعة 75 ملغم /کغم من وزن الجسم لوحظ احتقان الوعاء الدموی وتفجی سایتوبلازم الخلایا البنکریاسیة وتضخم بعضها فضلا عن زیادة الضرر النسجی عند الجرعة 75 ملغم /کغم. نستنتج من ذلک أن استخدام مضادات الاکتئاب قد یسبب زیادة الضغط التأکسدی النسجی فی الأعضاء الداخلیة للجسم خاصة إذا تم استخدامه خلال فترة الحمل بجرع عالیة، وبالنتیجة فان هذه الآفات لها تأثیر کبیر على صحة کل من الأجنة وأمهات الفئران الحوامل وبما أن اغلب الآفات المرضیة التی تم تسجیها کانت فی الأجنة والتی تسبب الإجهاض وبالتالی تؤثر على صحة أمهات الفئران الحوامل.

Introduction

Recently, the abuse of antidepressants such as fluoxetine is a significant problem worldwide (1). Those drugs are often used in depression treatment. They may help depressed people to recover from their sickness, especially pregnant women who suffer from depression and emotional changes during pregnancy and after childbirth (2). One of the newest antidepressant drugs is Fluoxetine Hydrochloride (3). The drug's chemical formula is C₁₇ H₁₈ F₃NO, and the scientific name of the drug is N-methyle-3-phenyle1-1-3 (4-trifluoromethyl) phenoxylpropan-1-amine (4). Fluoxetine metabolism is done in the liver, and the drug is metabolized into norfluoxetine. Norfluoxtine is the drug's active metabolite, which has been directly linked to medical and toxicological effects. It is the active metabolite of the drug directly linked to medical and toxicological effects (5). About 80% of the drug is excreted with urine, and 57% is excreted with feces (6). Moreover, the drug has a maternal effect. The severity of the effect depends on the dose and duration of exposure (7). Medical reports showed that the drug causes pulmonary hypertension syndrome in babies (8). Fluoxetine administration throughout gestation may indeed cause a slow formation of the lungs (9). The drug caused an increase in mortality and altering the structure of phospholipids, hepatic changes when given to rats at a dosage of 10,50 mg/kg b.w. (10). The drug induces oxidative stress, which causes testicular damage, a significant decrease in the weight of reproductive organs, and affects the level of testosterone in the mice (11). Drug administration also causes injuries in the vital organs of the body (12). Several studies referred to the drug's effect on carbohydrate metabolism and the function of beta cells (13).

The objective of this research project was to assess the impact of fluoxetine administration on the concentrations of 45,75 mg /kg of b.w. along with the maternal lung and pancreas of pregnant mice Mus musculus to evaluate the harmful effect of the drug on them.

Materials and methods

In our investigation, twenty-one pregnant mice, aged three months, weighted 29 ± 3 gm, were used. Animals were taken from the College of Veterinary Medicine's animal house, Mosul University, Mosul, Iraq, and housed in the animals' house of the Biology Department, College of Education for Pure Science. Animals were caged in plastic cages, supported with free access to food and water, fed with a standard diet, the room temperature was 25ºC, the animals were exposed to a regular light-dark cycle (14). The drug used in the study is fluoxetine 20 mg capsules produced by Bristol Laboratories Ltd., Bristol house, Unite 3, Canalside, Northbridge Road, United Kingdom. For the mating process, two females for one male were placed together in the same cage overnight. The next morning females with vaginal plugs were isolated in separate cages and kept together until the 7^th day of pregnancy. Animals’ housing was done following the standard guidelines for the use and care of experimental animals.

The pregnant mice (n=21) were divided into three groups (each group consisted of 7 pregnant mice). Group I (control group): The pregnant mice (n=7) were administered with 0.2ml of distilled water orally from the day 7^th until the 17^th of pregnancy. Group II: The pregnant mice (n=7) were administered orally with 45mg/kg b.w. of fluoxetine drug from the 7^th day until the 17^th day of pregnancy. Group III: in this group, the pregnant mice (n=7) were administered 75 mg/kg b.w. of fluoxetine drug from day 7^th until day 17^th of pregnancy.

Doses preparation

The drug solution was prepared by dissolving each concentration in 5 ml of distilled water (stock solution). The doses rates were between o.13 - 0.15 ml depending on pregnant mice weight. Those doses had been chosen depending on the LD₅₀ of the drug, which is 100 mg/kg b.w. (15). The doses (drug solution) were prepared freshly every day during the experiments period.

Histopathological preparation

On the day 17^th of pregnancy, all pregnant mice were sacrificed and dissected, maternal lung and pancreas were dissected. Specimens were fixed in formalin 10% for 48 hours; later, they were washed with distilled water for two hours and processed with routine paraffin embedding technique (16). Sections were stained with hematoxylin and eosin (17).

Results

Histopathological observations of the lung of adult pregnant mice

Light microscope examination of the control lung section of the pregnant mice Mus musculus showed normal appearance of lung histology (Figure 1). Lung sections collected from pregnant mice performed 45 mg/kg b.w. of fluoxetine from the 7^th until the 17^th day of pregnancy showed satisfactory Histopathological lesions represented with serofibrinous edema in the air space of alveoli and infiltration of inflammatory cells as well as enlarged airspace and mild thickening of alveolar septa (Figure 2) as well as degeneration of the pulmonary lining epithelium cells and congestion (Figure 3). While the examination of the lung sections obtained from pregnant mice given 75 mg/kg b.w. of fluoxetine drug orally for the same period above showed that previous lesions were increased, especially the congestion of blood vessel (Figure 4) as well as hyperplasia of the lung epithelium tissue was observed (Figure 5).

Figure 1: A cross-section photomicrograph of the lung of the control pregnant mouse Mus musculus showing normal lung histology. (10x. H&E).

Figure 2: A cross-section photomicrograph of the lung of pregnant mouse Mus musculus treated with fluoxetine drug at the dose of 45 mg/kg of b.w. from the 7^th day until 17^th day of pregnancy showing serofibrinous edema (1), enlargement of pulmonary air space (black arrow), mild thickening of alveolar septa (2), infiltration of inflammatory cells (3). (400x. H&E).

Figure 3: A cross-section photomicrograph of the lung of the pregnant mouse Mus musculus treated with fluoxetine drug at the dose of 45 mg/kg of b.w. from the 7th day until the 17th day of pregnancy showing degeneration of lung epithelium cells (4) and congestion (5). (10x. H&E).

Figure 4: A cross-section photomicrograph of the lung of the pregnant mouse Mus musculus treated with fluoxetine drug at the dose of 75 mg/kg of b.w. from day 7 to day 17 during pregnancy shows an increase in the congestion of blood vessels (7). (10x. H&E).

Figure 5. A cross-section photomicrograph of the lung of the pregnant mouse Mus musculus treated with fluoxetine drug at the dose of 75 mg/kg of b.w from day 7 to day 17 during pregnancy. showing hyperplasia of tracheal epithelium cells (black arrows). (10x. H&E).

Histopathological observations of the pancreas of pregnant mice

Light microscope examination of the control pancreas of pregnant mice Mus musculus showed normal lobules and normal pancreatic cells (Figure 6). Pancreas sections were obtained from pregnant mice given 45mg/kg b.w. of fluoxetine drug orally from the 7^th until the 17^th day of pregnancy. The histological changes and the dose increase were increased, so sections of the pancreas obtained from the pregnant mice given 75mg/kg b.w. of fluoxetine drug orally for the same period above revealed vacuolation of the pancreatic cells (Figure 7). Hypertrophy of some of them, congestion of the blood vessel, and the increase of eosinophilia of some pancreatic cells cytoplasm (Figure 8).

Figure 6: Cross-section photomicrograph of the pregnant mouse Mus musculus's control pancreas shows normal pancreatic lobules and normal pancreatic cells. (10x. H&E).

Figure 7: Cross-section photomicrograph of the pancreas of pregnant mice Mus musculus treated with fluoxetine drug at the dose of 75 mg/kg of b.w. from day 7 to day 17 during pregnancy showing vacuolation of pancreatic cells (black arrows). (10x. H&E).

Figure 8: Cross-section photomicrograph of the pancreas of pregnant mouse Mus musculus that treated with fluoxetine drug at the dose of 75 mg/kg of b.w. from day 7 to day 17 during pregnancy showing hypertrophy of some pancreatic cells (thin black arrow), eosinophils of some pancreatic cells cytoplasm (black arrows heads), congestion of the blood vessel (thick black arrow). (400x. H&E).

Discussion

In our study, the Macroscopical examination of the lung of pregnant mice administrated orally with 45,75 mg/kg b.w. of fluoxetine showed serofibrinous edema in the alveolar air space, degeneration of the alveolar lining epithelium, infiltration of inflammatory cells, congestion, and hyperplasia. The lesions were increased as well, as the dose increased. Those lesions were similar to those reported by (9,12,18). This study's findings correspond with those of (19) as all of them confirmed that taking fluoxetine during pregnancy causes hyperplasia of alveolar septa. The findings of this study may be directly attributable to that inflammatory response caused by fluoxetine. This induction may be linked to fluoxetine impairing the phospholipids metabolism, which involves some pathological lesions such as inflammatory cells infiltration and degeneration (20). on the other hand, it may be due to the use of fluoxetine during pregnancy, which may alter some metabolic pathways and increase in the levels of oxidative stress which in turn cause formation of destructive free radical, those molecules disturb and destroy the functions and structure of the lung cells (21).

The findings from the current study had shown histopathological lesions in pregnant mice's pancreas that were treated with fluoxetine at 45, 75 mg/kg b.w. for the preceding period, so the most noticeable lesions at the 75 mg/kg dose were blood vessel congestion, vacuolation of pancreatic cytoplasm and hypertrophy of some pancreatic cells. Study findings did not correspond with the findings of (22). At the same time, the present study results were similar to the finding of (23). The results were somewhat similar to the finding of (24). The finding of the current study may be due to the, that using fluoxetine for depression treatment during pregnancy may involve inducing cytotoxic mechanism in the body cells, which in turn increase the production of free radical oxygen species that destroy cellular mitochondria, proteins, lipids, and nucleic acids (25).

Conclusion

In conclusion, antidepressants may stimulate oxidative injury throughout the body's internal organs, particularly if taken at high doses during pregnancy. Consequently, they should always be used caution to avoid adversely.

Acknowledgements

Authors would express the thanks to University of Mosul, to support current study.

Conflict of interest

No conflict of interest.