Despite a general relation between fat intake and cardiovascular risk factors, the association is often not observed in studies conducted in heterogenic populations, as for population groups, it may differ. The aim of the study was to analyze the associations between dietary fat intake and lipoprotein cholesterol fractions, as well as atherogenic blood properties, in young and middle-aged overweight Caucasian women. In a group of 138 overweight women, the three-day dietary records were assessed, under-reporters were excluded, and lipoprotein cholesterol fractions were analyzed. For the included 24 young (aged 20–40) and 42 middle-age women (aged 40–60), the intakes of fat, saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), polyunsaturated fatty acids (PUFA), and cholesterol, as well as the PUFA/SFA ratio, were assessed. Afterwards, the analysis of associations with blood levels of total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), and triglyceride, as well as the TC/HDL ratio, HDL/LDL, ratio and Atherogenic Index of Plasma (AIP), were conducted. It was stated that the influence of the dietary fat level on lipoprotein cholesterol fractions as well as atherogenic blood properties in overweight Caucasian women is age dependent. For young, overweight, Caucasian women, the influence of the dietary fat level on the lipoprotein cholesterol fractions was not observed; however, SFA intake influenced atherogenic blood properties. For middle-aged, overweight, Caucasian women, the PUFA intake had an especially important influence in increasing the HDL cholesterol level. For overweight Caucasian women, not only should lipoprotein cholesterol fractions be controlled, but also the AIP calculated—especially for younger women.