Abstract <p>The aim of the work was to study changes in the content ofceramide and key enzymes involved in its biosynthesis, as well asTNFα content and TNFR1 expression levels, in the rat brain underconditions of acute cerebral ischemia (ACI) and hypoxic preconditioning(HP); additionally, we aimed to assess the relationship betweenthese factors and animal survival rates and neurological deficit.The experiments were carried out on 37 male outbred white rats weighing180–230 g. ACI was induced by irreversible ligation of the leftcommon carotid artery combined with reversible ligation of the rightcommon carotid artery. The animals were divided into three groups:group 1—sham-operated rats (control); group 2—rats exposed to ACI;and group 3—rats exposed to ACI and HP. On day 3 of observation,neurological deficit was assessed using the Garcia scale; changesin the levels of TNFα, TNFR1 expression, ceramide, serine palmitoyltransferase,ceramide synthase, and acid and neutral sphingomyelinases were evaluatedby immunofluorescence microscopy. ACI led to increased ceramideproduction and elevated levels of all the ceramide-synthesizingenzymes in the brain, being also accompanied by higher levels ofTNFα and upregulated TNFR1 expression. HP abated these effects,preventing the rise in ceramide, sphingomyelinase, and TNFα levels,as well as in TNFR1 expression, thereby reducing the severity of neurologicaldeficits and improving animal survival rates compared to ACI. Thus,HP demonstrated its efficacy as a method to reduce neurologicalimpairments and increase survival in experimental animals, as wellas to partly prevent changes in pro-inflammatory factors and ceramidelevels due to suppression of key enzymes involved in its synthesis.</p>