Caffeine and clomipramine are classified in pregnancy category C. The most critical time during pregnancy is the embryonic period [15]. Therefore, caution in prescription is necessary, particularly selleckchem Lapatinib at this time. There is evidence that coadministration of caffeine and clomipramine has a potentiating effect on caffeine toxicity [7]; so the aim of this study was to investigate the effects of simultaneous administration of different doses of these two drugs on fetal development in pregnant rats.2. Materials and Methods2.1. AnimalsAs the animal model, the Wistar-Albino strain of the laboratory rat inbred for 96 generations by brother sister litter-male mate mating was used. This strain was used for many years in our laboratory in Tehran University of Medical Science (2006-2007) and has never shown a prediction for spontaneous maternal defects.

Healthy adult female and male rats with an average age of approximately three month and weighing 300�C350 grams were randomly selected. They were kept in a controlled room (temperature, 20 to 25��C, humidity, 70% to 80%, exposed to 12h of daylight). The rats were fed with standard rat food and tap water until experimentation. Limitation of food and water was not applied to the animals that were put into their cages after the experiments. After mating, and ensuring successful conception, pregnant rats were divided into seven groups (n = 6). All experiments were conducted in Tehran University of Medical Sciences according to the recommendations of the ethics committee on animal’s experimentation of medical school.2.2.

DrugsIn all groups, predetermined doses of drugs were daily injected intraperitoneally between the eighth and fifteenth day of pregnancy. The first group was used as the control and received one mL of normal saline. In the second and third groups, clomipramine was injected in doses of 40mg/kg and 80mg/kg, respectively. Caffeine was injected at 60mg/kg and 120mg/kg to the fourth and fifth groups, respectively. Rats in group six received 40mg/kg clomipramine and 60mg/kg caffeine. Finally, the seventh group was administrated with clomipramine and caffeine at 80mg/kg and 120mg/kg, respectively. Drugs were purchased from Sigma-aldrich Company, USA. On the 17th day of pregnancy, the animals were anesthetized via inhalation of high concentrations of chloroform and the fetuses were removed by caesarean section.

2.3. Macroscopic and Microscopic StudiesThey were then examined for macroscopic abnormalities. Histopathological slides from fetuses were also prepared. After hematoxylin Drug_discovery and eosin staining, any microscopic changes in fetuses were noted using an optical microscope. Fetuses with abnormal body shape (non-C-shaped), subcutaneous hemorrhage, skin shrinkage, bent limbs, unilateral or bilateral cleft palates, and nonfused eyelids were considered abnormal [16].2.4. Statistical AnalysisData were analysed using statistical software SigmaPlot version 11.