Abstract: 【Abstract】  Objective：To preliminarily screened the protein molecules and biological processes that may be involved in PM2.5 increases the rate of adverse pregnancy by affecting the placental trophoblast cells using proteomic technology. Methods：HTR-8/SVneo cells were exposed to 0, 30, 60, 120 and 200 μg/mL PM2.5 for 24 h and 48 h, respectively, to observe the cell proliferation; finally, we selected 120 μg/mL PM2.5 treated with HTR-8/SVneo cells for 24 h and 48 h for follow-up experiments. After extraction of intracellular protein and enzyme digestion, peptides and differentially expressed proteins were analyzed and identified by isobaric tags for relative and absolute quantitation (iTRAQ) combined with two-dimensional liquid chromatography tandem mass spectrometry (2D LC-MS/MS) technology. Finally, bioinformatics analysis of the data was carried out. Results：The survival rates of cells treated with 120 μg/mL PM2.5 for 24 h and 48 h were 86.09% and 52.36%, respectively, which were also remarkably decreased compared with that of control cells (P＜0.05). There were 182 differentially expressed proteins in the cells exposure to 120 μg/mL PM2.5 for 24 h, involved in 22 cell biological processes (BP); meanwhile, cells exposed for 48 h had 486 differential proteins, involving 217 BP. Conclusions：PM2.5 can affect a large number of BP in HTR-8/SVneo cells. iTRAQ with LC-MS/MS 2D technology is an effective way for high sensitivity and throughput screening of differentially expressed proteins in cells after exposure to PM2.5, which provides a basis for the future research on the mechanism of PM2.5 increased the risk of adverse pregnancy outcomes.

Key words: 【Keywords】 , Proteome, Air pollutants, Tandem mass spectrometry, Trophoblast cells, Isobaric tags for relative and absolute quantitation.