ABSTRACT
Objective To investigate the expression of transforming growth factor-β1 (TGF-β1),endoglin (Eng) and their mRNA in placenta and superficial myometrium of patients with gestational hypertension or preeclampsia and to explore the role of Eng and TGF-β1 in the pathogenesis of gestational hypertension or preeclampsia.Methods One hundred and ten pregnant women were selected in the Second Affiliated Hospital of Tianjin Medical University from April 2009 to April 2010 who underwent cesarean sections and were divided into four groups:gestational hypertension group (n=30),mild preeclampsia group (n=30),severe preeclampsia group (n=30) and control group (normal pregnant women without labor and perinatal complications,n=20).The tissues of placenta and superficial myometrium were collected during cesarean section.Protein levels of Eng and TGF-β1 were detected by Western Blot.Real-time fluorescence reverse transcription polymerase chain reaction was used to detect the Eng and TGF-β1 mRNA expression.One-way ANOVA was used to compare among groups,Student-Newman-Keuls test was used to compare the differences between groups; relation between groups was analyzed by Pearson relation and linear regression.Results In control group,gestational hypertension group,mild and severe preeclampsia group,the Eng mRNA level was 1.00,1.27±0.58,1.54±0.41 and 1.83±0.35,and the TGF-β1 mRNA level was 1.00,1.64 ± 0.33,1.92± 0.38 and 2.23 ± 0.53 in placenta respectively; those figures changed to 1.00,1.32±0.46,1.59±0.37 and 1.93±0.52,and 1.00,1.71 ± 0.45,1.91 ± 0.51 and 2.37 ± 0.46 in superficial myometrium respectively.The Eng and TGF-β1 mRNA levels of control group were lower than those of the other three groups (P＜0.05).The higher the mRNA level,the more severe the disease (P＜ 0.05).In control group,gestational hypertension group,mild and severe preeclampsia group,the Eng protein expression was 0.11±0.07,0.15± 0.05,0.18 ± 0.06 and 0.43 ± 0.04,and the TGF-β1 protein expression was 0.11 ±0.02,0.26 ± 0.05,0.27± 0.03 and 0.88 ± 0.09 in placenta respectively; those figures changed to 0.14±0.06,0.16±0.04,0.20±0.08 and 0.46±0.05,and 0.15±0.03,0.29±0.06,0.31±0.04 and 0.91 ±0.08 in superficial myometrium respectively.The Eng and TGF-β1 protein levels of control group were lower than those of the other three groups (P＜0.05).The higher the protein level,the more severe the disease (P＜0.05).In the gestational hypertension group,mild and severe preeclampsia group,there were positive correlations between Eng and TGF-β1 protein levels in placenta (r=0.57,0.61 and 0.60 respectively,P＜0.05) and superficial myometrium (r=0.59,0.62 and 0.61 respectively,P ＜ 0.05).Conclusions Eng and TGF-β1 might play a role in pathogenesis of gestational hypertension and preeclampsia.
ABSTRACT
Objective To investigate the effects of soluble endoglin(sEng)on nitric oxide (NO)production and endothelial nitric oxide synthase(eNOS)phosphorylation in cultured human umbilical vein endothelial cells.Methods Human umbilical vein endothelial cells within 3 passages seeded in culture plates of 96 wells,were stimulated by total culture medium(control group)or sEng (1,10 and 100 μg/L)respectively.Cells and medium were collected after cells were cultured for 6,12 and 24 hours respectively.The concentration of the metabolites of NO in each group was measured by nitrate reductase method.The expression of eNOS and eNOS-Ser(p)1177 were detected by Western blot.The expression of eNOS mRNA in each group was detected by real-time fluorescence reverse transcription-polymerase chain reaction.Analysis of variance,LSD method and pearson correlation were used to compare the difference between groups.Results(1)The concentration of the metabolites of NO in 1,10 and 100μg/L sEng groups was(59.25±1.63),(41.08±2.71)and (30.38±1.63)μmol/L respectively after cultured for 6 hours;(54.98±3.34),(35.00±8.60)and (19.82±3.75)μmol/L for 12 hours; and(46.14±4.93),(30.24±2.08)and(12.78±5.01)μmol/L for 24 hours.There was no significant changes in control group with time going by(F=2.30,P=0.14).The concentration of the metabolites of NO was significantly lower in sEng group,and which had negative correlation with culture time(r=-0.98,P＜0.05)and dose(r=-0.88,P＜0.05).(2)The expression of eNOS in 1,10,100 μg/L sEng groups was 0.71 ± 0.00,0.47 ± 0.00 and 0.32±0.00 after cultured for 6 hours; 0.58±0.00,0.42±0.00 and 0.25±0.00 for 12 hours; and 0.49±0.00,0.33±0.00 and 0.18±0.00 for 24 hours.While the expression of eNOS and eNOS-Ser (p)1177/eNOS had no significant changes in control group with time going by(F=3.59 and 0.37,P=0.09 and 0.80).The expression of eNOS protein and eNOS-Ser(p)1177 decreased significantly in sEng groups,which had negative correlation with culture time(r=0.98 and-0.96,P＜0.05)and dose(r=-0.76 and-0.79,P＜0.05).(3)The expression of eNOS mRNA decreased significantly in sEng groups.Which also had negative correlation with culture time(r=-0.51,P＜0.05)and dose(r=-0.82,P＜0.05).Conclusions sEng might inhibit eNOS activity by blocking 1177 Ser phosphorylation to decrease NO production.