Diazepam-based drugs are widely used today in human treatment. Diazepam may be a primary drug aimed at treating neurological diseases or an associated drug in the treatment of other diseases in the purpose of symptomatic therapy. The sedative effect of diazepam characterizes it as a drug that people usually use on their own and without a doctor’s supervision. Directly, but also through influencing the nervous system, diazepam disrupts proper functioning of all body organs. The purpose of this paper was to examine the effects of diazepam on blood and cytohistological parameters of rats in an in vivo experiment. Mallory-Azan and immunochistochemical staining methods BLX-CX and Survivin tissues of liver, kidney and spleen of rats were used to achieve the set goal. Cytometric analysis of rats detected cells in apoptosis and measurements of stereological parameters were made using a system according to Cavalier’s principle. Results of analysis of hematological and histological parameters indicate a detrimental effect of diazepam on blood parameters, as well as on structure and functioning of the liver, kidneys and spleen of rats. This paper is a foundation for further detailed scientific research with the aim of elucidating all harmful effects that diazepam has on all organs in the body of rats. This data could serve as a starting point for future studies in clinical pharmacology on therapeutic protocols for usage of diazepam-based sedatives.