| Objective:The main purpose of this study is to detect the expression level of autophagy-related molecular markers in placental tissues of pregnant women with the decreased amniotic fluid volume and normal amniotic fluid,and to compare the differences between them,in order to explore the relationship between autophagy of placental trophoblasts and the decreased amniotic fluid volume in advanced stage pregnant,and to provide new ideas and directions for further revealing the pathogenesis of the decreased amniotic fluid volume in advanced stage pregnant.Methods:Thirty full-term parturients who were delivered by cesarean section in the first affiliated Hospital of Jinan University from January 2019 to December 2019 were selected as the study object,including 20 cases of amniotic fluid reduction as the experimental group and 20 cases of normal amniotic fluid as the control group.Transmission electron microscope was used to observe the occurrence of trophoblast autophagy in placental tissues of the two groups,immunohistochemistry was used to detect the expression of Beclin1,LC3 and p62 in placental tissues of the two groups,and Western blotting(Western Blot)and real-time fluorescence quantitative polymerase chain reaction(q RT-PCR)were used to detect the expression of Beclin1,LC3 and P62 in placental tissues of the two groups.Results:1.The results of transmission electron microscope showed that the content of autophagosomes in placental trophoblasts in the decreased amniotic fluid group was higher than that in normal amniotic fluid group.2.The results of immunohistochemical analysis showed that Beclin-1,LC3 and p62 were positive in the placental trophoblasts of the two groups,and the expression sites were mainly distributed in the cytoplasm of trophoblasts.The mean optical densities of Beclin-1,LC3 and p62 of placental trophoblasts in the decreased amniotic fluid group were 0.031±0.017,0.033 ± 0.016 and 0.008 ± 0.004 respectively,while those in normal amniotic fluid group were 0.019±0.009,0.021±0.014 and 0.013±0.009.The expression of Beclin-1 and LC3 in placental trophoblast in the decreased amniotic fluid group was significantly higher than that in normal amniotic fluid group,while the expression of p62 in oligohydramniosgroup was lower than that in normal amniotic fluid group.3.The results of q RT-PCR showed that the relative expressions of Beclin-1 and LC3 m RNA in placental tissue of the decreased amniotic fluid group were 4.133±1.577 and4.457±2.263 respectively,while those of normal amniotic fluid group were 2.458±1.686 and 2.531 ± 1.132.The expression levels of Beclin-1 and LC3 m RNA in the decreased amniotic fluid group were significantly higher than those in normal amniotic fluid group.4.The results of Western Blot showed that the relative expressions of Beclin-1 and p62 in placenta of the decreased amniotic fluid volume group were 2.962±0.565 and 0.803±0.436 respectively,while those in normal amniotic fluid group were 1.863±0.672 and 1.461± 0.245.The expression of Beclin-1 in the decreased amniotic fluid volume group was significantly higher than that in normal amniotic fluid group,while the expression of p62 in the decreased amniotic fluid volume group was lower than that in normal amniotic fluid group.Conclusion:The autophagy activity in the placenta of the decreased amniotic fluid volume in advanced stage pregnant is enhanced,suggesting that the increase of autophagy level is closely related to the occurrence and development of the decreased amniotic fluid volume in advanced stage pregnant.The excessive autophagy of placental trophoblasts may be one of the reasons for the decreased amniotic fluid volume in advanced stage pregnant. |
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