16(S)-HETE

Abstract

Many researchers suggest an increased risk of atherosclerosis in women with polycystic ovary syndrome. In the available literature, there are no studies on the mediators of inflammation in women with PCOS, especially after dietary intervention. Eicosanoids (HETE and HODE) were compared between the biochemical phenotypes of women with PCOS (normal and high androgens) and after the 3-month reduction diet. Eicosanoid profiles (9(S)-HODE, 13(S)-HODE, 5(S)-HETE, 12(S)-HETE, 15(S)-HETE, 5(S)-oxoETE, 16(R)-HETE, 16(S)-HETE and 5(S), 6(R)-lipoxin A4, 5(S), 6(R), 15(R)-lipoxin A4) were extracted from 0.5 ml of plasma using solid-phase extraction RP-18 SPE columns. The HPLC separations were performed on a 1260 liquid chromatograph. No significant differences were found in the concentration of analysed eicosanoids in phenotypes of women with PCOS. These women, however, have significantly lower concentration of inflammatory mediators than potentially healthy women from the control group. Dietary intervention leads to a significant (p < 0.01) increase in the synthesis of proinflammatory mediators, reaching similar levels as in the control group. The development of inflammatory reaction in both phenotypes of women with PCOS is similar. The pathways for synthesis of proinflammatory mediators in women with PCOS are dormant, but can be stimulated through a reduction diet. Three-month period of lifestyle change may be too short to stimulate the pathways inhibiting inflammatory process.

2.5.2. Sample Preparation

Venous blood was collected from the patients after an overnight fasting. Whole blood was collected and placed in tubes containing ethylenediaminetetraacetic acid (EDTA) as anticoagulant. The blood was immediately placed on ice or in a refrigerator, and next the samples were centrifuged at 3500 rpm for 10 min at 4°C within max 2 h of collection. Plasma was stored at −80°C. Standard blood biochemical analyses were performed in the University Hospital Laboratory. 5(S), 6(R)-lipoxin A4, 5(S), 6(R), 15(R)-lipoxin A4, 5(S)-HETE, 5(S)-oxoETE, 12(S)-HETE, 15(S)-HETE, 16(R)/16(S)-HETE, 9(S)-HODE, and 13(S)-HODE were extracted from 0.5 ml of plasma using a solid-phase extraction RP-18 SPE columns (Agilent Technologies, Cheadle, UK) []. Recovery and sensitivity of method were described previously []. The total recovery for all sample extraction and processing steps (mean ± SD) was 46 ± 8%.

3. Results

When comparing the concentrations of analysed inflammatory state mediators between the group of women with PCOS (PCOS-I) and the control group (CG), it was observed that statistically significant differences were present in levels of almost all eicosanoids except lipoxins A4 (, ). Similar relation with respect to the control group (CG) was observed when the patients were divided into two phenotypes: with high (HT) and normal (NT) levels of androgens. When we compared two phenotypes of women with PCOS (HT and NT) to each other, we did not observe any statistically significant differences. However, an increasing tendency was noted in the group of women with normal level of androgens (NT) (). After the three-month reduction diet with lower glycemic index, a significant increase in the majority of inflammatory factors was observed, except for 6(R), 15(R)-lipoxin A4 (). Women with PCOS (PCOS-II) after being three months on the diet did not differ significantly from the control group (CG), however, the levels of 12(S)-HETE were much higher ().