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Several reports demonstrated that mice exposed to LPS at middle gestational stage caused neural tube defects(NTDs).The present study investigated the effects of vitamin D3 supplementation on LPS-induced NTDs.Pregnant mice except controls were intraperitoneally(i.p.)injected with LPS(20 μg/kg)daily from gestational day(GD)8 to GD12.In the LPS+VitD3 group,pregnant mice were orally administered with vitamin D3(25 μg/kg)before LPS injection.As expected,a five-day LPS injection resulted in 62.5%(10/16)of litters and 27.5%of fetuses with NTDs.Additional experiment showed that a five-day LPS injection down-regulated placental proton-coupled folate transporter(pcft)and reduced folate carrier(rfc),two major folate transporters in placentas.Consistent with down-regulation of placental folate transporters,folate transport from maternal circulation into embryos was disturbed in the LPS group.Interestingly,vitamin D3 supplementation not only inhibited LPS-evoked inflammation but also inflammation-associated down-regulation of placental folate transporters.Correspondingly,vitamin D3 counteracted inflammation-associated perturbation of folate transport from maternal circulation into embryos.Importantly,vitamin D3 protected against LPS-induced NTDs.Taken together,these results suggest that vitamin D3 supplementation prevents LPS-induced NTDs through improving placental folate transportation.Thus,maternal vitamin D3 supplementation during pregnancy may have potential utilities for preventing inflammation-associated NTDs.