Amitriptyline is one of the first reference tricyclic antidepressants (TCAs) with sedative and analgesic properties. This study aimed to investigate the testicular indices that influenced by amitriptyline treatment and examined it by biochemical, histological and Immunohistochemical methods. A total of 20 male albino rats were equally divided into two groups: the first was control and the second was amitriptyline intoxicated group. The results of this study showed that amitriptyline consumption significant decrease hormones parameters, increase sperms abnormalities, decrease sperm numbers, induced testicular tissue damage and P53 alterations.