1 The Relationship Between Expression Of Aquarorin-8 And Aquarorin-9 In Human Fetal Membrane And Placenta Of Patients With Gestational Diabetes Mellitus And The Changes Of Amniotic Fluid Volume 2 Effects Of Indomethacin On Aquaporin8 Expression In The Hum

ⅠTHE RELATIONSHIP BETWEEN EXPRESSION OF AQUARORIN-8 AND AQUARORIN-9 IN HUMAN FETAL MEMBRANE AND PLACENTA OF PATIENTS WITH GESTATIONAL DIABETES MELLITUS AND THE CHANGES OF AMNIOTIC FLUID VOLUMEObjective: To explore the relationship between the expression of aquarorin-8 and aquarorin-9 in human fetal membrane and placenta of patients with gestational diabetes mellitus and the changes of amniotic fluid volume, and to investigate the function of aquaporin-8 and aquaporin-9 in regulating amniotic fluid volume of patients diagnosed as gestational diabetes mellitus. So as to provide some new clues for the treatment of GDM polyhydramnios.Methods: Twenty cases of GDM complicated with polyhydramnios, twenty cases of GDM with normal volume of amniotic fluid and twenty cases of normal term pregnancy were recruited and divided into three groups. Fetal membrane (amnion and chorion) and placenta samples from the patients mentioned above were taken after they delivered with cesarean sections. The expression of quarorin-8 and aquarorin-9 mRNA and protein in fetal membrane(amnion and chorion) and placenta tissues were dectected with semi-quantitive RT-PCR and Western blotting.Results: The expression of AQP8 mRNA in amnion, chorion and placenta of GDM polyhydramnios group was (0.72±0.085), (0.62±0.131), and (1.18±0.090) respectively, and that of control group was (0.47±0.041), (0.56±0.085), and (0.82±0.096) respectively. The expression of AQP8 mRNA in amnion, chorion and placenta of GDM with normal volume of amniotic fluid group was (0.50±0.068), (0.61±0.120), and (0.84±0.074) respectively. The expression of AQP8 protein in amnion, chorion and placenta of GDM polyhydramnios group was (0.54±0.044), (0.47±0.112), and (0.90±0.098) respectively, and that of control group was (0.32±0.052), (0.43±0.058), and (0.49±0.039) respectively. The expression of AQP8 protein in amnion, chorion and placenta of GDM with normal volume of amniotic fluid group was (0.34±0.031), (0.46±0.075), and (0.51±0.058) respectively. In amnion and placenta, the expression of AQP8 mRNA and protein of GDM polyhydramnios group was increased compared to that of control group and GDM with normal volume group (P<0.05), but in chorion the difference in AQP8 mRNA and protein expression between the two of the three groups was not significant (P>0.05). In amnion and placenta samples, the difference in AQP8 mRNA and protein levels between the GDM with normal volume of amniotic fluid group and the control group was not significant (P>0.05).The expression of AQP9 mRNA in amnion, chorion and placenta of GDM polyhydramnios group was (0.79±0.106), (0.60±0.132), and (0.87±0.087) respectively, and that of control group was(0.49±0.094), ( 0.55±0.096),and (0.61±0.044) respectively. The expression of AQP9 mRNA in amnion, chorion and placenta of GDM with normal volume of amniotic fluid group was (0.51±0.072), (0.58±0.058), and (0.63±0.062) respectively. The expression of AQP9 protein in amnion, chorion and placenta of GDM polyhydramnios group was (0.59±0.042), (0.48±0.121), and (0.79±0.051) respectively, and that of control group was (0.28±0.031), (0.42±0.102), and (0.47±0.047) respectively. The expression of AQP9 protein in amnion, chorion and placenta of GDM with normal volume of amniotic fluid group was (0.30±0.061), (0.44±0.096), and (0.48±0.039) respectively. In amnion and placenta, the expression of AQP9 mRNA and protein of GDM polyhydramnios group was increased compared to that of control group and GDM with normal volume group (P<0.05), but in chorion the difference in AQP9 mRNA and protein expression between the two of the three groups was not significant (P>0.05). In amnion and placenta, the difference in AQP9 mRNA and protein expression between the GDM with normal volume of amniotic fluid group and the control group was not significant (P>0.05).Conclusion: In both amnion and placenta tissues, expression level of quarorin-8 and aquarorin-9 mRNA and protein of GDM complicated with polyhydramnios group were markedly up-regulation than GDM with normal volume of amniotic fluid group and normal control group. The aquarorin-8 and aquarorin-9 in both amnion and placenta tissues may play key roles in regulating amniotic fluid volume of GDM. ⅡEFFECTS OF INDOMETHACIN ON AQUAPORIN8 EXPRESSION IN THE HUMAN AMNIOTIC EPITHELIAL CELLSObjective: Indomethacin is the medicine for primary polyhydramnios, and the study discuss the effects of indomethacin on aquaporin8 expression in the human amniotic epithelial cells, so as to provide some evidences to the treatment of polyhydramnios, and provide a new target to the drug treatment of polyhydramnios.Methods: Amniotic epithelial cells were primary culture through isolated from human amniotic membrane.Cell types are characterized with immuocytochemistry. Amniotic epithelial cells were divided into experimental groups and normal control groups. The experimental groups were fed with indomethacin of different concentration(10,20,50,100, 200μM/L)for 24 hours,and in other groups the amniotic membrane cells were fed with indomethacin 100μM/L for 6h,12h,24h,36h and 48h respectively. The changes of AQP8 mRNA and protein levels in amniotic membrane cells treated by indomethacin were assayed by RT-PCR and Western blotting.Results: Different concentrations of indomethacin decreased the expression of AQP8 mRNA and protein in experimental groups. The levels of AQP8 mRNA and protein reached the peak and significantly decreased compared with the control groups with 200μM/L indomethacin (P<0.01).The expression of AQP8 mRNA and protein in experimental groups decreased obviously when the amniotic membrane cells were fed with 100μM/L indomethacin (P<0.01). And with the longer treated time, the expression of AQP8 mRNA and protein decreased much.Conclusion: Indomethacin could down-regulation the expression of AQP8 mRNA and protein in the human amniotic epithelial cells.