[Study on the oxidative stress and inflammation in trophoblast cells stimulated by different chain length fatty
Xiao-le Sun1, Zi Yang, Xiao-Ye Wang
1Department of Obstetrics and Gynecology, Peking University Third Hospital, Beijing 100191, China.
Objective:
To investigate the oxidative stress and inflammation in trophoblast cells stimulated by different chain length fatty acids.
Methods:
Serum-free trophoblast cells cultured in vitro were divided into five groups, which were incubated with DMEM medium without free fatty acid (F-FFA), short chain fatty acids (SC-FFA), medium chain fatty acids (MC-FFA), long chain fatty acids (LC-FFA), very long chain fatty acids (VLC-FFA). Then cells in each group were stimulated by DMEM medium, reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor (apocynin) and p38 mitogen-activated protein kinases (p38MAPK) inhibitor (SB203580) and were subdivided as each FFA plus-DMEM group, plus-NADPH-I and plus-p38MAPK-I groups. Expressions of mRNA and protein of p38MAPK and cyclooxygenase 2 (COX-2) in trophoblast cells were detected by real-time PCR and western blot.
Results:
(1) The mRNA expression of p38MAPK in LC-FFA + DMEM, VLC-FFA + DMEM, LC-FFA + NADPH-I, LC-FFA + p38MAPK-I, VLC-FFA + NADPH-I, VLC-FFA + p38MAPK-I group were 4.56 ± 0.28, 22.65 ± 2.40, 0.87 ± 0.06, 1.02 ± 0.15, 19.87 ± 1.93, 10.22 ± 0.75 separately, and the protein expressions were 0.79 ± 0.02, 0.93 ± 0.10, 0.43 ± 0.06, 0.44 ± 0.19, 0.79 ± 0.10, 0.81 ± 0.14. Compared with other groups, the mRNA and protein expressions of p38MAPK in LC-FFA + DMEM, VLC-FFA + DMEM group were increased (P < 0.05). Compared with LC-FFA + DMEM group, mRNA and protein expressions of p38MAPK in LC-FFA + NADPH-I and LC-FFA + p38MAPK-I group were significantly decreased (P < 0.05). Compared with VLC-FFA + DMEM group, mRNA and protein expressions of p38MAPK had no difference in VLC-FFA + NADPH-I group (P > 0.05), mRNA expression of p38MAPK in VLC-FFA + p38MAPK-I group was significantly decreased (P < 0.05), but there was no difference in protein expression (P > 0.05). (2) The mRNA expression of COX-2 in LC-FFA + DMEM, VLC-FFA + DMEM, LC-FFA + NADPH-I, LC-FFA + p38MAPK-I, VLC-FFA + NADPH-I, VLC-FFA + p38MAPK-I group were 3.97 ± 0.03, 39.08 ± 0.63, 0.99 ± 0.13, 0.98 ± 0.18, 20.93 ± 3.70, 13.46 ± 2.31 separately, and the protein expressions were 1.32 ± 0.20, 1.33 ± 0.25, 0.59 ± 0.13, 0.58 ± 0.30, 0.88 ± 0.18, 0.91 ± 0.24. Compared with other groups, mRNA and protein expressions of COX-2 in LC-FFA + DMEM and VLC-FFA + DMEM group were significantly increased (P < 0.05). Compared with LC-FFA + DMEM group, mRNA and protein expressions of COX-2 in LC-FFA + NADPH-I and LC-FFA + p38MAPK-I group were decreased (P < 0.05). Compared with VLC-FFA + DMEM group, mRNA and protein expressions of COX-2 in VLC-FFA + NADPH-I and VLC-FFA + p38MAPK-I group were all decreased (P < 0.05). (3) The correlation analysis showed that there were significantly positive correlations between the mRNA and protein expressions of p38MAPK and COX-2 in LC-FFA group (P < 0.05). There were significantly positive correlations in protein expression (P < 0.05), but no correlation in the mRNA expression between p38MAPK and COX-2 in the F-FFA, SC-FFA, MC-FFA, VLC-FFA groups (P > 0.05).
Conclusions:
The oxidative stress and inflammation may exist in trophoblast cells which were stimulated by LC-FFA and VLC-FFA. p38MAPK signal transduction pathway may contributed in this process.
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