同型半胱氨酸干扰滋养细胞线粒体并影响细胞融合功能OA

Objective:To investigate the effect of homocysteine(Hcy)on the fusion process of trophoblast cells and its poten-tial mechanism.Methods:Bewo cells were cultured in vitro to mimic in vivo trophoblasts,successfully constructing a cell fusion model with forskolin(FSK),and treating cells with 50 µmol/L Hcy for 48 hours.A confocal microscopy was used to observe the fusion status and calculate the fusion index,and the human chorionic gonadotropin(hCG)levels were detected by ELISA.The mRNA expression levels of fusion genes ERVW-1,ERVFRD-1,GCM1,and CGB,as well as the mRNA expression of genes relate to mitochondrial function and oxidative stress were detected by qPCR.The protein expression levels of ERVW-1,ERV-FRD-1,and GCM1 were measured by Western blot.Intracellular reactive oxygen species(ROS)levels and membrane potential were detected by immunofluorescence.ATP levels were measured using chemiluminescence method.The activity of superoxide dismutase(SOD)was determining by WST method.Results:Trophoblast cells were successfully fused under FSK induction,and the cells showed significant changes upon the addition of Hcy;the cell fusion index was significantly reduced(P<0.001);the hCG secretion was greatly reduced(P<0.001),and both gene and protein expression levels relate to cell fusion were significantly down-regulated(P<0.05).Mitochondrial membrane potential declined(P<0.001),the ATP levels significantly decreased(P<0.01),and the expression of mitochondrial fusion-related genes substantially reduced(P<0.001),while the expression of mito-chondrial fission genes widely increased(P<0.05).The amount of intracellular ROS production was significantly increased(P<0.01).The activity of T-SOD was visibly declined(P<0.001),and the mRNA expression level of NRF2,a key gene of oxida-tive stress was significantly decreased(P<0.001).Conclusion:Hcy may have a negative impact on placental development and pregnancy health by interfering with the normal function of mitochondria,promoting the production of ROS,triggering oxidative stress,and hindering the normal fusion of trophoblasts and hCG secretion.