Lentivirus-driven Hematopoietic Prostaglandin D Synthase Overexpression Exerts a Negative Regulatory Effect on the Corpus Luteum in Sheep

The maintenance and regression of corpus luteum are regulated by prostaglandin F2α, but the role of prostaglandin D2 (PGD2) has not been thoroughly investigated.Its synthesis involves the key enzyme, hematopoietic prostaglandin D synthase (HPGDS) that mediates PGD2 production in different cell types.The aim of this study was to investigate the biological role of endogenous PGD2 on luteal cells in Hu sheep via HPGDS overexpression.For this purpose, luteal cells obtained from luteal tissue of 5 healthy adult Hu sheep at the mid-luteal phase immediately after slaughter were isolated via mechanical dissociation, enzymatic digestion, and Percoll density gradient centrifugation, cryopreserved in liquid nitrogen, and retrieved for subsequent experiments.Subsequently, quantitative real-time PCR (qRT-PCR) was used to confirm the efficiency of HPGDS overexpression.ELISA, qRT-PCR and Western blot (WB) were utilized to detect key indicators of luteal function and structural regression.Immunofluorescence was used to localize the PGD2 receptors.Analytical results of lentiviral construction showed that HPGDS lentiviral plasmids were constructed, packaged in 293T cells, with MOI of 160 determined as optimal for luteal cell infection.HPGDS overexpression reduced P4 concentration and StAR expression (P