Eotaxin is a chemokine, which is a chemoattractant mainly to eosinophils, as well as basophils and Th2 lymphocytes. According to studies, overexpression of eotaxin is found in endothelial and smooth muscle cells of blood vessels in the area of atherosclerotic plaque. In clinical medicine, cardio-ankle vascular index (CAVI) is widely used as an indicator of arteriosclerosis and a predictor of cardiovascular events. Few studies have shown the relationship of eotaxin with coronary atherosclerosis; in other studies, the relationship of eotaxin with atherosclerosis, myocardial infarction and pulse wave velocity was not revealed. The aim of the present study was to assess blood level of eotaxin and cardio-ankle vascular index and their association with major cardiovascular risk factors in patients with high and very high cardiovascular risk. We examined 65 patients with high and very high cardiovascular risk, due to documented coronary artery disease, type 2 diabetes mellitus, or combination of cardiovascular risk factors and who were undergoing generally accepted cardioactive, hypoglycemic therapy and lipid-lowering therapy. All patients were examined for the elastic properties of the vascular wall by volumetric sphygmography with assessment of CAVI. In the blood, the concentrations of eotaxin, high-sensitivity C-reactive protein, glycosylated hemoglobin and lipid spectrum indicators were determined. All examined were divided into two groups: with a normal value of CAVI (less than 8) and elevated. Patients with elevated CAVI had higher concentrations of eotaxin (p = 0.013), total cholesterol (p = 0.009), low-density lipoprotein cholesterol (p = 0.016), were older (p < 0.0001) and less likely to take statins (p = 0.002). In all those examined, correlations were found between serum eotaxin concentration and CAVI (rs = 0.34; p = 0.005), as well as age (rs = 0.32; p = 0.006). The age of the patients correlated with CAVI (rs = 0.35; p = 0.007). Thus, in our study, we for the first time showed the relationship between higher concentrations of eotaxin and an increased cardio-ankle vascular index in patients with high and very high cardiovascular risk. Cardio-ankle vascular index was associated with age, lipid metabolism and lipid-lowering therapy. The obtained results allow us to consider eotaxin as a factor associated with atherogenesis and arterial stiffness.