An increasing level of evidence supports the utility of pravastatin as prevention against preeclampsia (preE). We previously demonstrated a hyperglycemia induced cytotrophoblast (CTBs) dysfunction characteristic of a preE-like phenotype and sought to demonstrate the utility of pravastatin in rescuing CTBs from this hyperglycemia induced dysfunction.
Human CTBs were treated with 100, 150, 200, 300, or 400mg/dL glucose for 48 hrs. Cells were treated with pravastatin (1ug/mL) either alongside or 2 hrs prior to glucose exposure. Some cells were treated with D-Mannitol as a negative control for glucose exposure. Cell migration was performed by Matrigel migration assay kit according to manufacturer protocol. Cell lysates were utilized to evaluate the mRNA expression of urokinase plasminogen activator (uPA) and plasminogen activator inhibitor 1 (PAI-1), while also assessing proliferating cell nuclear antigen (PCNA) and p38 MAPK phosphorylation by western blot. Levels of vascular endothelial growth factor (VEGF), placental growth factor (PlGF), soluble fms-like tyrosine kinase-1 (sFlt-1), soluble endoglin (sEng) and interleukin 6 (IL-6) were measured in culture media using ELISA kits. Statistical comparisons were performed using analysis of variance with Duncan's post hoc test.
Hyperglycemia inhibited CTBs migration by down-regulating uPA, PAI-1, PCNA and up-regulating p38 phosphorylation in cells treated with >150mg/dL glucose compared to basal (100mg/dL) (*p<0.05 for each). Secretion of sFlt-1, sEng and IL-6 were increased while VEGF and PIGF were decreased in CTBs treated 150mg/dl of glucose (*p<0.05 for each). Both pravastatin pretreatment and co-treatment significantly rescued CTBs migration, up-regulating uPA, PAI-1, PCNA, down-regulating p38 phosphorylation, and correcting the angiogenic profile of CTBs (p<0.05 for each). D-Mannitol showed no osmotic effect on CTBs.
Pravastatin mitigates the hyperglycemia-induced dysfunction of CTBs by attenuating the glucose-induced anti-proliferative, anti-migratory, anti-invasive and anti-angiogenic phenotype similar to that seen in Preeclampsia. This study supports the potential for pravastatin use on CTBs development early in pregnancy and the importance of continuing research of pravastatin in preE prevention.