The Effects Of UDCA And SAMe On The Expression Of OATP1B3in Human Trophpblast Cells

Background:Intrahepatic cholestasis of pregnancy (ICP), a specific complication in the second half of human pregnancy, have no effective protective and therapeutic measures been discovered so far. The significant characteristic of ICP is the rise of bile acid level in maternal blood、cord blood、amniotic fluid and meconium. It is strongly suggested that placenta may take part in bile acids transport between pregnancy woman and fetus. It has revealed that Organic anion transport polypeptide1B3(OATP1B3) expressed in placenta and can efficiently transport bile acids. Our study shows that OATP1B3was down-regulated in placenta of ICP contrast to normal group and it plays an important role in transport of GCA and GCDCA in human placenta.Many researchs revealed that OATP1B3take great part in bile acids transport. We investigated the influence of UDCA and SAMe on the expression of OATP1B3.Our results may contribute to the characteristics of bile acids transport in human trophoblast cells and fetal risk prevention.Objectives:To investigate weather UDCA and SAMe effects on the expression of organic anion transporting polypeptides1B3(OATP1B3). To provide a potential strategy for reducing the fetal risk and a novel therapeutic target for ICP treatment. Methods:Use MTT method to detect the cytotoxicity of two treatment drug of ingtrahepatic cholestasis of pregnancy (ICP). The human trophoplast cell line were treated with UDCA and SAMe for different concentration and different time. The expression of mRNA and protein were analyzed by realtime PCR and Western blotting.Results:(1) MTT method:UDCA in the concentration of25μM or50μM had no cytotoxicity to SWAN cell within72h (P>0.05); SAMe in the concentration of5μM、10μM、15μM、20μM、35μM had no cytotoxicity to SWAN cell within72h(P>0.05);(2) UDCA and SAMe effects on the expression of OATP1B3mRNA by Real-time PCR method:a.After treated SWAN cell with25μM、50μM UDCA for24h: OATP1B3mRNA expression increased14%、20%respectively(P>0.05); The mRNA of OATP1B3expression increased43%after treated SWAN cell with25μM UDCA for48h (P>0.05),50μM UDCA treated group increased66%(P<0.05); b. After treated with SWAN cell15μM,35μM SAMe for24h:OATP1B3mRNA expression increased83%、320%(P<0.05)respectively(P<0.05);48h later, mRNA of OATP1B3expression increased48%、191%respectively (P<0.05); Results Significant difference between15μM SAMe treated group and35μM SAMe treated group (P<0.05).(3) UDCA and SAMe effects on the expression of OATP1B3protein level by Western blot method:a. After treated with SWAN cell20μM、50μM UDCA for48h:OATP1B3protein expression increased1%,146%respectively, Results Significant difference between the normal groups and50μM UDCA treated group (P<0.05), with statistical significance;b. After treated with SWAN cell15μM、35μM SAMe for48h:OATP1B3protein expression increased31%、28%respectively (P<0.05), these two groups had a marked difference (P<0.05)Conclusions:(1) UDCA with the concentration no more than50μM have no cytotoxicity within72h; SAMe with the concentration no more than35μM have no cytotoxicity within72h;(2) UDCA and SAMe can increase mRNA expression of OATP1B3,both trugs have a concentration and time effect.(3) UDCA and SAMe can increase the protein expression of OATP1B3in human trophoblast cells.