Fang Zheng, Mao Jiaqin, Chen Shuqiang, Dong Jie, Wang Xiaohong
Reproductive Medicine Center, Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi'an 710038, China.
Reproductive Medicine Center, Department of Obstetrics and Gynecology, Tangdu Hospital, Air Force Medical University, Xi'an 710038, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2022 Jun;38(6):535-541.
Objective To identify the effect of HLA-G-containing exosomes on the secretory function of growth-promoting factors osteoglycin (OGN) and pleiotrophin (PTN) by decidual NK (dNK) cells. Methods dNK cells were co-cultured with HLA-G-containing exosomes from the villi of patients undergoing unexplained recurrent pregnancy loss (uRPL) and normal induced abortion, respectively. Sequentially, OGN and PTN of the dNK cells were determined using real time quantitative PCR and western blotting. Exosomes overexpressing HLA-G (HLA-G-EXO) were obtained by transfecting the villous trophoblast cell line HTR-8/Svneo with lentivirus LV-HLA-G. dNK cells were further co-cultured with HLA-G-EXO for detecting the expression of OGN and PTN, the culture supernatant of which was used to treat HTR-8/Svneo cells, and the proliferation of HTR-8/Svneo cells was detected by the CCK-8 assay. Results Exosomes derived from villi of patients receiving normal induced abortion significantly enhanced the expression of OGN and PTN in dNK cells compared with those from patients of the uRPL group. Besides, HLA-G-EXO markedly enhanced the expression of OGN and PTN in dNK cells. The culture supernatant of HLA-G-EXO treated dNK cells could promote the proliferation of HTR-8/Svneo cells. Conclusion Villi-derived HLA-G containing exosomes may enhance the secretion of growth-promoting factors in dNK cells.
目的 确定含HLA-G的外泌体对蜕膜自然杀伤(dNK)细胞分泌促生长因子骨形成蛋白(OGN)和多效生长因子(PTN)功能的影响。方法 将dNK细胞分别与不明原因复发性流产(uRPL)患者绒毛及正常人工流产患者绒毛中含HLA-G的外泌体共培养。随后,采用实时定量聚合酶链反应和蛋白质免疫印迹法检测dNK细胞中OGN和PTN的表达。通过慢病毒LV-HLA-G转染绒毛滋养层细胞系HTR-8/Svneo获得过表达HLA-G的外泌体(HLA-G-EXO)。将dNK细胞与HLA-G-EXO进一步共培养以检测OGN和PTN的表达,其培养上清液用于处理HTR-8/Svneo细胞,采用细胞计数试剂盒-8法检测HTR-8/Svneo细胞的增殖。结果 与uRPL组患者相比,正常人工流产患者绒毛来源的外泌体显著增强了dNK细胞中OGN和PTN的表达。此外,HLA-G-EXO显著增强了dNK细胞中OGN和PTN的表达。经HLA-G-EXO处理的dNK细胞培养上清液可促进HTR-8/Svneo细胞的增殖。结论 绒毛来源的含HLA-G的外泌体可能增强dNK细胞中促生长因子的分泌。