Evaluation of Embryonal Protective Effect of Melatonin against Apoptotic Effect of Methadone in Embryo Tissues of Mice

Introduction: Methadone is a substance that is widely used in the substitution treatment of opiate addiction. This study aimed to evaluate the protective effects of melatonin on apoptosis induced by transfer of transplacental methadone in mice. Methods: This study was an experimental study aimed to assay apoptotic effect of placenta transferred methadone on brain, liver and kidney tissues and to study protective effect of melatonin in an embryo model in mice.  Bulb-C mice weighing 25-30 g were kept in four shelves (3 females and one male) and checked daily for the sterile cycle. For mice with vaginal plugs, the first day of pregnancy was considered. After confirmation of pregnancy, female mice were divided into 5 groups of 6, which included the control group, methadone (10 mg / kg) and melatonin groups in three doses (2, 4, 6 mg / kg / day) by IP injection for half an hour before prescribing methadone. This prescription was given for 10 days from the beginning of pregnancy. After the infants were born, their liver, brain, and kidneys were removed and they underwent immunohistochemical tests for apoptotic expression of Bcl2, BAX, and Caspase3 proteins. The results were analyzed by SPSS version 22 statistical software and Chi-square (Chi 2) and Fisher tests (P <0.01). Results: This study showed that melatonin at dose 4 and 6mg/kg has decreased apoptotic protein BAX and Caspase9 and increased anti-apoptotic protein, Bcl2 expression approximately, but these results were not significant statistically (P>0.05). In addition, for dose of 0.2 mg/kg of melatonin, there was not any apoptotic effect. Conclusion: Our findings showed that melatonin has almost a protective effect against apoptotic effect induced by placental transfer of methadone especially via its increasing effects on Bcl2, but this anti-apoptotic effect was not significant and needed for more precise studies.