Aim <p>The purpose of this study was to discuss effects of exosome miRNA-369-3p on the biological behavior of trophoblast cells in gestational diabetes.</p> Materials and methods <p>Measuring miRNA-369-3p mRNA expression in Normal and GDM pregnant women by RT-qPCR assay; using high glucose to make GDM cell model; using CCK-8 and flow cytometry to evaluate cell proliferation andapoptosis; measuring cell invasion and migration by transwell and wound healing assay; IL-10 mRNA and protein expression by RT-qPCR and WB assay; correlation between miRNA-369-3p and IL-10 by Dual luciferase assay.</p> Results <p>Compared with normal pregnant women, miRNA-369-3p mRNA level significantly up-regulation in GDM pregnant women (<i>P</i>&lt;0.001); with high glucose stimulating, compared with Normal group, the cell proliferation rate (78.30 ± 4.27%) significantly inhibited with cell apoptosis significantly increasing (24.55 ± 2.01%) (<i>P</i>&lt;0.001), cell invasion (65.33 ± 3.65%) and migration (48.31 ± 4.18) activities significantly inhibited with IL-10 significantly down-regulation (<i>P</i>&lt;0.001, respectively). With miRNA-369-3p inhibitor supplement, the cell activities were significantly improved with IL-10 increasing.</p> Conclusion <p>Exosome miRNA-369-3pknockdown had effects to improved high glucose induced HTR-8/Svneo cell biological activities by stimulating IL-10.</p>