Study on the oxidative stress and inflammation in trophoblast cells stimulated by different chain length fatty acids / 中华妇产科杂志

ABSTRACT

Objective To investigate the oxidative stress and inflammation in trophoblast cells stimulated by different chain length fatty acids.MethodsSerum-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 conrelation in the mRNA expression between p38MAPK and COX-2 in the F-FFA,SC-FFA,MC-FFA,VLC-FFA groups (P ＞ 0.05).ConclusionsThe 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.