| Objectives To investigate the expression of miRNA-155(miR-155)and Angiotensin II receptor 1(AGTR1)in placenta of patients with preeclampsia and make sure the correlation of them.Methods 1 A total of 65 cases of pregnant women who underwent cesarean section in Tangshan Maternal and Child Health Hospital and Tangshan Workers’ Hospital from November 2019 to July 2020 were collected,including 29 cases in normal pregnancy group,15 cases in preeclampsia group and 21 cases in severe preeclampsia group.Realtime quantitative fluorescence PCR(RT-PCR)was used to detect the expression of miR-155 in the placental tissues of each group.Western Blotting and Immunohistochemistry were used to detect the expression of AGTR1 protein in the placental tissues of each group.2 miR-155 was transfected into JEG-3 cells which were cultured by ourselves for overexpression.The cells were divided into blank group,Lip group,miR-NC group and miR-155 mimics group.The transfected efficiency was detected by Fluorescence labeling and RT-PCR.After confirmed the successful construction of the cell model,RT-PCR and Western Blotting were used to detect the expression of AGTR1 m RNA and AGTR1 protein in each group and compared their differences.MTT and Transwell tests were used to detect the proliferated and invased ability of JEG-3 cells which had transfected with miR-155.3 The SPSS 22.0 software was used for statistical analysis of the experimental data and the measurement data was expressed as(x ± s).One-way ANOVA was used for the comparison of data between multiple samples,LSD and SNK tests were used for the comparison of data between two samples,Rank and inspection were used for processing count data and Pearson analysis was used for correlated analysis(α=0.05).P<0.05 means that the difference is statistical significant.Results 1 In the normal group,preeclampsia group and severe preeclampsia group,the expressed amount of miR-155 were(1.181±0.116),(1.573±0.072),(1.981±0.096)respectively,they presented an increased trend and the difference between them was statistical significant(F=384.234,P<0.001).The expressed amount of AGTR1 protein in the three group were(0.845±0.071),(0.579±0.033),(0.303±0.042)respectively,they presented a decreased trend and the difference between them was statistical significant(F=584.627,P<0.001).2 FAM-has-miR-155 was transfected into JEG-3 cells for overexpressed.The fluorescence of FAM in the cells were observed under fluorescence microscope which showed the transfected efficiency was about 80% and these cells could be used for subsequent experiments.In the blank group,Lip group,miR-NC group and miR-155 mimics group,the expressed amount of miR-155 were(0.927±0.129),(1.241±0.090),(1.617±0.095),(4.251±0.432)respectively,the amount in miR-155 mimics group was increased obviously higher than other three groups,the difference between them was statistical significant(F=125.712,P<0.001),if blank group,Lip group and miR-NC group were compared with each other,the difference between them were no statistical significant(P=0.140,0.069,0.086,P>0.05).In these four groups,the expressed amount of AGTR1 m RNA were(0.988±0.064),(0.995±0.041),(1.032±0.085),(0.435±0.119)respectively,the amount in miR-155 mimics group was decreased obviously lower than other three groups,the difference between them was statistical significant(F=35.919,P<0.001),if blank group,Lip group and miR-NC group were compared with each other,the difference between them were no statistical significant(P=0.923,0.536,0.599,P>0.05).The expressed amount of AGTR1 protein in the four groups were(1.549±0.031),(1.563±0.017),(1.549±0.029),(0.786±0.018)respectively,the amount in miR-155 mimics group was decreased obviously lower than other three groups,the difference between them was statistical significant(F=732.330,P<0.001),if blank group,Lip group and miR-NC group were compared with each other,the difference between them were no statistical significant(P=0.487,0.972,0.508,P>0.05).3 MTT and Transwell tests showed that the proliferated and invased ability of JEG-3 cells which were transfected by miR-155 were significantly inhibited in the miR-155 mimics group that compared with other three groups(P<0.05),if blank group,Lip group and miR-NC group were compared with each other,the difference between them were no statistical significant(P>0.05).Conclusions 1 In the placental tissues,the expression of AGTR1 could be regulated by miR-155,they are correlated negatively and both of them play an important role in the progression of preeclampsia.2 In the placental tissues of patients with preeclampsia,the expression of miR-155 is increasing and AGTR1 is decreasing,the change of both of them are further obvious with the aggravated condition of preeclampsia.3 In the transfected JEG-3 cells,the overexpressed miR-155 could inhibit the expression of AGTR1 m RNA and AGTR1 protein,in the mean time,it could inhibit the ability of proliferation and invasion.4 miR-155 and AGTR1 could act as the influence factors for intervention and diagnosis of preeclampsia.Figure 18;Table 16;Reference 104... |
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