The aim of this work was to study the expression of Toll-like receptors (TLRs) genes in the hypothalamic structures of the brain, after the application of acute stressful effects. A hypothesis has been put forward about Toll-like receptors as a key link in the mechanisms of the implementation of a stress reaction, including at the level of the central nervous system (CNS). The important role of TLR in the pathogenesis of stress-mediated diseases of the central nervous system is assumed. The expression of TLR2, TLR3, and TLR4 genes in rat hypothalamus was studied for after 3 hours after the application of acute emotional-physical stress. A reliable increase in the level of gene expression of all three receptors at the mRNA level in stressed animals was established compared with the control. The obtained experimental data indicate the activation of the system of Toll-like receptors at the level of the central nervous system under stress. The activation of several receptors of the TLR family with different specificities in the absence of an increase in microbial load, including at the level of pathogen-associated molecular factors (PAMP), may also indicate the likely significant role of endogenous TLR ligands in the described processes.