The role of clomipramine in potentiating the teratogenic effects of caffeine in pregnant rats: a histopathological study

Abstract

Since little is known about the teratogenic effects of clomipramine used concurrently with caffeine during the organogenesis period, the aim of this study was to test the teratogenic effects of a coadministration of caffeine and clomipramine on rat fetuses. We divided 42 pregnant rats into seven groups, randomly. The first group (control) received 0.5 mL of normal saline. Clomipramine was injected at 40 mg/kg and 80 mg/kg to the second and third groups, respectively. The fourth and fifth groups received caffeine in doses of 60 mg/kg and 120 mg/kg, respectively. The sixth group received a combination of 40 mg/kg clomipramine and 60 mg/kg caffeine, and the seventh group was given clomipramine and caffeine at 80 mg/kg and 120 mg/kg, respectively. The fetuses were removed on the 17th day of pregnancy and studied in terms of microscopic and macroscopic morphological features. Fetuses of rats receiving high doses of caffeine or combinations of caffeine and clomipramine showed a significant rate of cleft palate development, open eyelids, mortality, torsion anomalies, shrinkage of skin, and subcutaneous haemorrhage (P ≤ 0.001). This study concludes that caffeine in high doses or the simultaneous administration of caffeine and clomipramine leads to teratogenicity.

Figure 1

Macroscopic view of 17-day-fetuses from control group (a) and the group that received a combination of clomipramine and caffeine in high doses (b). In the control group fetus, the muzzle is formed normally. The body is C-shaped and eyes, ears, and upper and lower extremities are in their normal locations. The skin is taut (a). In the fetus from the group treated with clomipramine and caffeine in high doses, the body is not fully C-shaped, the forelimbs are bent, and corrugated skin and areas of subcutaneous hemorrhage are obvious (b).

Figure 2

Histopathological slides of the frontal section of the head, 17-day-fetuses in the control group (a) and the group that received a combination of clomipramine and caffeine in high doses (b) (H&E staining, 4x). In the control group (a), the nasal septum is attached to the roof of the mouth and nostrils (A) are completely separated from the oral cavity. Tongue (B) is shown to be perfectly in the mouth and located in its normal place. In the fetus from the group treated with clomipramine and caffeine in high doses (b), there is bilateral clefting of the palate (C) and tongue inside the mouth is flat (D).

Figure 3

Histopathological slides of the eyes from 17 days fetuses in control group (a) and the group received a combination of clomipramine and caffeine in high doses (b) (H&E staining, 10x). In control group fetus (a), the upper (A) and lower (B) eyelids are joined completely. In the fetus from the group treated with clomipramine and caffeine in high doses (b), the upper (C) and lower (D) eyelids were separated completely and the cornea (E) is exposed.