| Research backgroundThe placenta is an important organ for exchanging substances between the fetus and the mother.The fetus develops in the uterus by relying on the placenta to obtain nutrition from the mother,and the placenta also synthesizes various hormones,enzymes and cytokines to maintain normal pregnancy.The formation of the placenta is the basis for the normal development of the embryo.During functional placenta development,migration and invasion of trophoblast cells play an important role.The process is strictly regulated by the body.Studies have shown that trophoblast cell migration and invasion play an important role in the development of gestational diseases such as early abortion,hypertensive disorder complicating pregnancy(HDCP)and fetal developmental delay.However,the regulation mechanism of trophoblastic migration and invasion is still unclear.Therefore' studying its molecular mechanism can provide new ideas for the diagnosis and treatment of gestational diseases.Exosomes(EXOs)are microscopic vesicles that mediate the transmission of material and information between cells through various pathways,secrete from the cell to the extracellular environment through exocytosis,and are stably present in the extracellular environment.There are great differences in the components of exosomes from different cell sources,so it is considered as an important local microenvironmental control medium and participates in various physiological processes of pregnancy during pregnancy.At present,it has been found that placenta-derived exosomes exist in the maternal circulation and regulate the functional changes of the maternal systemic organs during pregnancy to meet the needs of pregnancy.If abnormalities occur in this regulation process,pregnancy disorders may occur.Mesenchymal stem cells(MSCs)are adult stem cells derived from the early stage of mesoderm development with strong self-repairing and multi-differentiation potential.They were first discovered in the bone marrow,and subsequently in the fat,umbilical cord,muscle,liver,etc.A variety of tissues and organs are also found in the mesenchyme.Studies have shown that there are a large number of mesenchymal stem cells in the uterus.Mesenchymal stem cells in the uterus can secrete a large number of cytokines,play an important role in endometrial angiogenesis,and dysfunction of uterine mesenchymal stem cells and the occurrence of preeclampsia.closely related.Mesenchymal stem cells can generate paracrine secreted biologically active substances--exosomes,and participate in a variety of pathophysiological processes including tissue damage repair,angiogenesis,and promotion of tumor microenvironment construction.Recent studies have found that tumor-derived exosomes can release a series of cytokines,growth factors,adhesion molecules and extracellular matrix proteins,mediating the tumor microenvironment and tumor cell-cell communication.Myelin-derived mesenchymal stem cell-derived exosomes can promote the migration and invasion of tumor cells and enhance tumor angiogenesis.For example,hepatoma cells,colon cancer cells,and exosome produced by melanoma cells can promote the invasion of corresponding tumors.In the process of placenta formation,embryonic trophoblast cells invade into the mother's aponeurosis and its spiral arteries,remolding the uterine spiral arteries in place of vascular endothelial cells to establish an interactive cycle between the mother and the fetus to provide nutrition for embryonic development.Its cell migration and invasion are similar to those of tumor invasion.At present,the correlation and regulation mechanism of mesenchymal stem cell-derived exosomes in trophoblastic migration have not been reported yet.Therefore,further exploration of the relevance of mesenchymal stem cell-derived exosomes and trophoblastic migration is necessary to reveal its relevance.The potential role in the diagnosis and treatment of gestational diseases has important clinical significance.Research purposesThis study will initially investigate the correlation between mesenchymal stem cell-derived exosomes and trophoblastic migration,thereby further revealing its pathogenesis in pregnancy-related diseases.Research methods1.Exosomes were extracted from mesenchymal stem cells C3H10T1/2 by ultracentrifugation and purified again by centrifugation.2.The size and morphology of exosomes were analyzed by TEM and Nanosight.3.The expression level of exosome surface-ALIX+TSG101 was detected and identified by SDS-PAGE and Western blot.4.Using PKH26 experiments confirmed the fusion of exosomes with trophoblast HTR8/Svneo.5.The effects of exosomes derived from mesenchymal stem cells at different concentrations(25ug/ml,50ug/ml)on migration of HTR8/Svneo cells were investigated by cell migration assay.6.The expression of MMP-2 and cell migration-related gene MMP-2 in trophoblast HTR8/Svneo fused with exosomes was detected by RT-PCR.Result1.The sediment of mesenchymal stem cell supernatant obtained by ultra high speed centrifugation using transmission electron microscopy and Nanosight was observed to be round,with a size of 40-100 nm;Western blot further confirmed its expression of exosome-specific markers.-ALIX,TSG101.2.The trophoblast cell HTR8/Svneo could uptake mesenchymal stem cell-derived exosomes.Compared with 6h and 12h,the uptake was higher at 24h.3.Transwell experiments showed that exosomes derived from mesenchymal stem cells can significantly promote the migration of trophoblasts,and the higher the concentration,the more significant the migration.4.After uptake of mesenchymal stem cells-derived exosomes,the expression of trophoblast HTR8/Svneo MMP2 gene was significantly up-regulated and correlated with its concentration.ConclusionsMesenchymal stem cell-derived exosomes can significantly promote the migration of trophoblast HTR8/Svneo,and its mechanism may be caused by up-regulation of HTR8/Svneo MMP2 gene expression. |
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