Cytokines appeared to contribute to the development of pathologic condition and eNOS gene polymorphism may affect cytokine production. The aim of this study was to evaluate cytokines pattern in preeclampsia and whether there is any relationship between gene and cytokines production and cytokine with disease severity. This cross-sectional study included 100 women with preeclampsia and 100 healthy pregnant women. Their blood samples were analyzed for nitric oxide (NO), inflammatory cytokines, and eNOS gene polymorphism. Decreased NO and increased cytokine (tumor necrosis factor-α, interleukin-2, and interferon-γ) levels were found in preeclampsia (p < 0.001). Significant differences were found in genotype/allele distribution between the two groups. A significant negative correlation was observed between NO and cytokine levels (tumor necrosis factor-α and interferon-γ) in the preeclamptic group (p = 0.001). We conclude that preeclampsia is associated with decreased levels of NO and increased levels of circulating inflammatory cytokines due to single nucleotide polymorphisms, pointing toward the role of endothelial and inflammatory components.