Effects of interfering FABP4 gene on lipid accumulation and migration of endometrial epithelial cells in sheep
The purpose of this study was to investigate the effect of FABP4 gene on lipid accumulation and migration of endometrial epithelial cells in sheep.Three FABP4 interference fragments were designed and ligated with pGBplv-U6-mcherry-puro to construct interference vectors.The empty plasmid was used as the control(NC group),and the three interference recombinant plasmids were FABP4-shRNAl,FABP4-shRNA2,and FABP4-shRNA3,respectively.The recombinant plasmids were identified by enzyme digestion.After plasmid transfection,RT-qPCR was used to identify the interference efficiency.Western Blot was used to further verify the interference effect of the vector.The vector was transfected into endometrial epithelial cells cultured in vitro.The accumulation of lipid droplets was detected by BODIPY method.The triglyceride content in the cells was detected by detection kit.RT-qPCR was used to detect genes related to lipid metabolism,proliferation and migration.Cell scratch test was used to detect cell migration.The results showed that all vectors can effectively interfere with the expression of the FABP4 gene,with the FABP4-shRNA2 vector showing the best interference effect.After interfering with the expression of the FABP4 gene in endometrial epithelial cells,compared with the NC group,the ability of cells to aggregate lipid droplets decreased,the content of triglycerides was reduced,the expression of mRNA,related to cellular lipid metabolism and proliferation and migration decreased,and the speed of scratch healing was reduced.After reducing the expression of the FABP4 gene in endometrial epithelial cells,the level of cellular lipid accumulation and the ability of cells to migrate decreased.The study indicates that the FABP4 gene may play an important role in the pregnancy of sheep and the normal development of embryos,and it is worth further in-depth study in breeding practice.
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