96 Inflammatory stress and deposition markers in the serum and on adipose tissue in normal and hypertensive pregnancy

Abstract

Introduction Not only in hypertensive pregnancy but also in normal course, homeostatic inflammation on adipocyte and adipose tissue has been confirmed by basic pilot study. We previously showed inflammatory and anti-inflammatory genetic action of the adipose tissue under the new culture method in preeclamptic serum compared with normal pregnancy (Naruse K, et al. Mediators Inflamm, 2015). Objectives In this study, we measured Interleukins, proteases and cell death markers on the inflammatory pathway, in the culture supernatants of visceral adipose tissue with normal and preeclamptic serum. Additionally, other markers for lipid oxidative stress and adipose storage were measured in the serum from normal pregnancy and preeclampsia. Methods All samples were taken under informed consent of the patients. Subcutaneous fat tissue from repeated cesarean section and omentum (visceral fat) from probe laparotomy for ovarian cancer were dissected, vessels were removed and captured in Peptide Hydrogel (BD Biosciences, Inc.). After 24 h starvation, tissues were cultured additional 24 h with medium containing 10% of maternal serum from severe preeclampsia (n = 5) and gestational age-matched healthy pregnant maternal serum (n = 5). Ratio of M30 (Apoptosis marker)/M65 (all cell death marker including necrosis) protein were measured with ELISA after correction with original serum concentrations of the molecules. Concentration of LDH (cytotoxicity marker) was measured with specific assay. Interleukin (IL)-6 and Matrix Metalloproteinase (MMP)-2 were measured with SearchLightR Multiplex Immunoassay. On the serum samples taken from 1st, 2nd and 3rd trimester of normal pregnancy and at onset of severe preeclampsia (n = 10 each), Malondialdehyde (MDA: a lipid marker for oxidative stress) were measured with Thiobarbituric Acid Reactive Substances (TBARS) Assay and cell death-inducing DFFA-like effector c (CIDEC/FSP27: lipid storage and adipocyte apoptosis marker) were measured with ELISA. Results In adipose tissue culture, M30/M65 ratio showed trend to increase in preeclampsia than normal pregnancy (p = 0.053). However, LDH was significantly increased in normal pregnancy than preeclampsia (p Conclusions In the adipose tissue, increase of apoptosis was shown after addition of preeclamptic serum. On the other hand, total tissue injury seemed to be stronger in normal pregnancy than preeclampsia, suggesting that adipose tissue act as not only an inflammatory focus but as a systemic buffer for acute inflammation in severe preeclampsia. Inflammatory stress on adipose tissue in normal pregnant subjects and preeclampsia were shown firstly by MDA concentrations. Taken together, adipose tissue is proposed as a key organ of variated pathophysiology of preeclampsia.