| Objective To study the expression and localization of Caspase-3, Caspase-8, Caspase-9 in human amniochorionic membrane, then investigate the role of Caspase-3, Caspase-8, Caspase-9 in human premature rupture of fetal membranes.Methods Amniochorionic membranes were collected from the following groups of women: (1) women with spontaneous premature rupture of fetal membranes (PROM) (n=8), (2)women with normal labor in term after vaginal delivery (n=8) (comparative group), (3)women undergoing elective repeat cesarean section (C-section) before the onset of labor and who had no complications of pregnancy (n=8) (comparative group). Caspase-3, Caspase-8, Caspase-9 peptides were studied with use of S-P. The morphologic features of fatal membranes was studied with optical microscope, the ultrastructure of fetal membrane cell was studied with electron microscope.Results (1)The expression of Caspase-3 and Caspase-9 peptides expressed in three groups. (1)The localization Caspase-3 and Caspase-9: They mainly stained in the amniotic epithelial cells and chorionic cytotrophoblast cells, Minority stained in amnion mesenchymal cells and cells of the reticular of the matrix, and which all presenced in cell plasma. (2)The expression of Caspase-3 and Caspase-9 peptides: Caspase-3 peptides, PROM group was [(62.86±3.83) ×10~-2 ], vaginal delivery group was [(42.33 ±2.99)×10~-2], C-section group was [(20.97±2.94) ×10~-2], There were statistically significant changes among the three groups (P<0.05); Caspase-9 peptides, PROM group was [(52.20 ± 3.28) × 10~-2], vaginal delivery group was [(25.87 ± 5.52) × 10~-2], C-section group was [(17.65 ±1.78) ×10~-2], The expression of Caspase-3 and Caspase-9 peptides concentration were highest in PROM group, followed by normal labor groupand C-section group, There were statistically significant changes among the three groups (P<0.05). (2)Caspase-8 almost was negative in three groups.(3)In PROM, the morphologic features of fatal membranes changed significantly in optical microscope, There were large numbers of apoptosis cells In electron microscope.Conclusion Caspase-9 and Caspase-3 respective as start-up apoptosis gene and apoptosis effective gene expressed more in PROM than comparative group, There were large numbers of apoptosis cells In electron microscope too. Large numbers of human fetal membranes cells apoptosis, So we suggest that Caspase-3 and Caspase-9 may play a role in the pathogenesis of PROM.Objective To study the expression and localization of MMP-2, MMP-9 and their inhibitors TIMP-2, TIMP-1 mRNA and peptide in human amniochorionic membrane, then investigate the role of MMP-2, MMP-9,TIMP-2 and TIMP-1 in human premature rupture of fetal membranes.Methods Amniochorionic membranes were collected from the following groups of women: (Dwomen with spontaneous premature rupture of fetal membranes (PROM) (n=8), (2) women with normal labor in term after vaginal delivery (n=8) (comparative group), (3)women undergoing elective repeat cesarean section (C-section) before the onset of labor and who had no complications of pregnancy (n=8) (comparative group). Messenger ribonucleic acid (mRNA) expression for MMP-2, MMP-9, TIMP-2 and TIMP-1 were studied with use of reverse transcriptase-polymerase chain reaction (RT-PCR). MMP-2, MMP-9, TIMP-2 and TIMP-1 peptides were studied with use of S-P.Results (l)The expression of MMP-2 mRNA and peptide: MMP-2 mRNA, PROM group was [(84.92+3.68) X10"2], vaginal delivery group was [(32.65+2.34) X10"2], C-section group was [(30.65+2.77) X 10"2]; MMP-2 peptide: PROM group was [(73.36+ 2.62) X10"2], vaginal delivery group was [(54.71 +4.16) X 10"2], C- section group was [(53.90+3.03)X10"2]. The expression of MMP-2 mRNA and peptide in PROM was higher than other groups. There were statistically significant changes (P<0.05). The expression of MMP-2 mRNA and peptide in vaginal delivery group and C- section group had no statistically significant change (P>0.05). (2)MMP-9 mRNA and peptide: MMP-9 mRNA , PROM group was [(2.60+0.37)X 10"2], vaginal delivery group was [(0.76+0.05) X 10"2]; MMP-9 mRNA was absent in C-section. MMP-9 peptide: PROM group was [(67.59+ 1.98)X 10"2], vaginal delivery group was [(53.07+1.53)X10"2], C-section group was [(43.19 ± 2.74) X 10'2]. The expression of MMP-9 mRNA and peptides concentration were highest in PROM group, followed by normal labor group and C-section group, there were statistically significant changes among the three groups CPO.05). (3)TIMP-2 mRNA and peptide: TIMP-2 mRNA, PROM group was [(42.01 ± 12.17)X 10"2], vaginal delivery group was [(73.01 ± 14.82) X 10"2], C- section group was [(88.47+6.51) XIO"2]; TIMP-2 peptide: PROM group was [(45.13+2.82) X 10"2], vaginal delivery group was [(60.80+4.35) X 10"2], C- section group was [(81.87 +4.26) X 10'2]. The expression of TIMP-2 mRNA and peptides concentration were highest in fetal membranes from C-section, followed by normal labor and PROM, There were statistically significant changes among the three groups (PO.05). (4)TIMP-1 mRNA and peptide: TIMP-1 mRNA, PROM group was [(44.15+2.00) XI0'2], vaginal delivery group was [(43.12+1.55) X10"2], C- section group was [(42.73 + 1.06) X 10~2]; TIMP-1 peptide: PROM group was [(56.99+1.71) X 10"2], vaginal delivery group was [(56.87+1.29) X10"2], C- section group was [(55.59+1.65) X10'2]. The expression of TIMP-1 mRNA and peptides in three groups has no statistically significant change CP>0.05). (5)The presence of MMP-2, MMP-9, TIMP-2 and TIMP-1: They expressed both in the amniotic epithelial cells and chorionic cytotrophoblast cells, and all presenced in cell plasma. TIMP-2 and TIMP-1 expressed in cells of the reticular of the matrix also, whereas the amnion mesenchymal cells produced only MMP-2 and very little MMP-9.Conclusion In PROM, MMP-2 and MMP-9 mRNA and peptide expressed increased. MMP-2 and MMP-9 were Activated from different way, which can increase they decomposing ECM. The imbalance of MMP-2/ TIMP-2 and MMP-9/ TIMP-1 may increase the role. The both reasons may cause weakening of the membranous elasticity and tenacity, then leading to rupture.
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