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Objective To investigate the correlation of the expressions of advanced glycation end products(AGE) and the receptor for advanced glycation end products(RAGE) in serum and placenta with the pathogenesis of preeclampsia. Methods From December 2013 to June 2014, 32 women with severe preeclampsia who received cesarean section in the Affiliated Hospital of Qingdao University were recruited in the study, defined as the severe preeclampsia group. 30 healthy pregnant women who received cesarean section in the same hospital were recruited as the control group. ELISA was used to measure the maternal serum AGE, soluble receptor for advanced glycation end products (sRAGE) and tumor necrosis factor-α(TNF-α) in these women. Furthermore, ELISA was also used to measure AGE and TNF-α in the placenta. The localizations of AGE and RAGE protein in placentas were detected by immunohistochemical SP method. RAGE and TNF-α mRNA expression in placentas were measured by real-time quantitative PCR. AGE, RAGE and TNF-αprotein expression in placentas were measured by western blot, respectively. Results (1) The serum levels of AGE,sRAGE and TNF-αin the severe preeclampsia group were (538 ± 75),(367 ± 86) and (322 ± 40) ng/L,respectively. They were significantly higher than those in the control group[(454 ± 50), (286 ± 35) and (270 ± 35) ng/L, respectively](P0.05). (2) In the severe preeclampsia group, the levels of AGE and TNF-αin placentas were (500 ± 82) and (334 ± 57) ng/L, which were higher than those in the control group [(431 ± 74) and (263 ± 46) ng/L, respectively](P<0.05). The levels of AGE showed positive correlation with the levels of TNF-ɑ(r=0.406,P<0.05). (3)AGE and RAGE protein mainly located in the syncytiotrophoblasts, macrophages and vascular endothelial cells in the placentas of the two groups. AGE expressed mainly in the cytoplasm, and RAGE expressed in the cytoplasm and cell membranes.(4)RAGE and TNF-αmRNA expression in the placentas of the severe preeclampsia group were 12.6 ± 4.6 and 10.4 ± 2.4, which were significantly higher than those in the control group (0.9 ± 0.4 and 3.5 ± 0.9,P<0.01). (5) The expressions of AGE、RAGE and TNF-αprotein in placentas of the severe preeclampsia group were 0.68 ± 0.06, 0.82 ± 0.08 and 0.76 ± 0.08. All were significantly higher than those of the control group (0.46 ± 0.05,0.42 ± 0.09 and 0.52 ± 0.07;P<0.01). Conclusions The levels of AGE and RAGE in serum and placentas elevated in the severe preeclampsia group, and the expression of TNF-αalso elevated. These indicated that AGE and RAGE might be involved in the systemic inflammatory response and local inflammatory response in placentas, and then caused the preeclampsia.
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Objective To research the correlation of the expressions of lipocalin-2 (LCN-2) and its receptor (NGALR) in serum and placenta with preeclampsia.Methods From Dec.2010 to Apr.2011,64 women with preeclampsia who delivered in Affiliated Hospital of Qingdao University Medical College were recruited in the study,including 26 women with moderate preeclampsia ( MPE group) and 38 women with severe preeclampsia (SPE group).Twenty-five healthy pregnant women were taken as control group.LCN-2 and NGALR mRNA and protein expression in placenta were measured by reverse transcription-PCR ( RTPCR) and western blot,respectively.Results ( 1 ) The serum levels of LCN-2 in MPE group and SPE group [ (58 ±20),(90 ± 18) μg/L] were significantly higher than that in control group [ ( 19 ±6) μg/L,P＜0.01] ; the serum LCN-2 level in SPE group was significantly higher than that in MPE group (P ＜0.01).(2) LCN-2 mRNA expression in placenta in MPE group and SPE group (0.55 ±0.14,0.61 ±0.14) were both significantly higher than that in control group (0.28 ±0.16,P ＜0.01 ) ; LCN-2 protein expression in placenta of MPE group and SPE group ( 2.2 ± 0.4,2.4 ± 0.5 ) were also significantly higher than that in control group (1.4 ±0.4,P ＜0,01 ),no significant difference was found between MPE group and SPE group ( P ＞ 0.05 ),(3) No significant difference was found in the expressions of NGALR mRNA in placenta among MPE group,SPE group and control group (0.46 ±0.1l,0.46 ±0.14,0.45 ±0.15,P ＞0.05 ).(4) NGALR protein expressions in MPE group,SPE group and control group were 2.7 ±0.8,3.0 ±0.9,and 2.7 ± 0.9,and there were no significant difference among these three groups ( P ＞ 0.05 ).(5) In preeclampsia,serum LCN-2 level significant associated with 24 hours total urinary protein and uric acid ( r =0.565,0.476,P＜0.01).LCN-2 serum level were not associated with systolic pressure and diastolic pressure (P ＞ 0.05 ) ; there were no association with the expressions LCN-2 mRNA aud protein in placenta ( P ＞ 0.05).Conclusions Serum LCN-2 level is closely related to the progress of preeclampsia.Increasing expression of LCN-2 in placenta may be a compensatory response to preeclampsia.
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Objective To investigate the variance levels of plasma soluble leukocyte differentiation antigens CD40 (sCD40) and soluble CD40 ligand (sCD40L) in preeclamptic patients with renal damage and its relationship. Methods A total of 63 pregnant women attended the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College between August 2008 and June 2010. In the present study included 28 pregnant women with mild preeclampsia and 35 patients with severe preeclampsia. Thirty matched normotensive pregnant women were enrolled in the study as the control group. Expression of sCD40 and sCD40L were determined by ELISA. At the same time, the blood routine, C reaction protein ( CRP),urine routine, 24 hours urine protein excretion, and serum uric acid (UA), creatinine (Cr), blood urea nitrogen (BUN) were measured. The correlation analysis was performed between the sCD40/sCD40L and the blood biochemical indexes in 3 groups. Results ( 1 ) The median levels of CRP in severe preeclampsia (10. 8 mg/L)and mild preeclampsia group(7. I mg/L)are significantly higher than that of control group (3. 3 mg/L,P ＜ 0. 05 ); The level of CRP in severe preeclampsia group was also higher than that of mild preeclampsia group ( P ＜ 0. 05 ). The median gestational age at delivery in severe preeclampsia ( 32. 5 weeks)was significantly less than that of mild preeclampsia group ( 37. 2 weeks) and normal group ( 38. 6 weeks,P ＜ 0. 05). However no significant differences were observed between mild preeclampsia group and normal group ( P ＞0. 05 ). The platelet count in severe preeclampsia ( 132 × 109/L) was significantly less than those of mild preeclampsia group (212 × 109/L) and normal group ( 216 × 109/L, P ＜ 0. 01 ), but no significant differences were observed in blood platelet amount between mild preeclampsia group and normal group ( P ＞0. 05 ). There was no significant difference in hemoglobin level and white blood cell in three groups ( P ＞0. 05). (2) The sCD40 plasma concentration in severe, mild preeclampsia and normal group was 133.6,126. 5 and 90. 7 ng/L, respectively. The sCD40 L plasma concentrations were 12. 5, 10. 4 and 4. 4 ng/L respectively in the 3 groups. 24 hours urinary protein quantitative was 4. 5 g/d,0. 8 g/d and 0 in the 3 groups respectively. And the UA level was 486 μ mol/L,289 μmol/L and 162 μmol/L. In the above three groups,the monitoring indicators were significantly higher in women with severe preeclampsia group compared with mild preeclampsia and control groups (P ＜ 0. 01 ), and there were also higher in mild preeclampsia group than that in control groups ( P ＜ 0. 01 ). The level of plasma Cr ( 89 μmol/L) and BUN ( 5. 32 mmol/L) in severe preeclampsia group were higher than those of mild preeclampsia group (66 μmol/L and 4. 49mmol/L) and control group ( 57 μmol/L and 3.32 mmol/L, P ＜ 0. 05 ). There was no significant difference between mild preeclampsia group and normal group (P ＞ 0. 05 ). (3) The correlation analysis indicated that the level of sCD40 has a positive correlation with 24 hours urinary protein quantitative( r = 0. 434, P ＜ 0. 05 ),also significant positive correlation( r =0. 536,0. 528 ,P ＜ 0. 01 ) between the level of sCD40 and UA or CRP in women with preeclampsia. There was no significant correlation between the level of sCD40 and systolic blood pressure, diastolic blood pressure, delivery gestational age, Cr, BUN, and platelet count(r =0. 135,0. 183, -0. 133,0. 190,0. 167, -0. 221 ,all P ＞0. 05 ). There were positive correlation between the level of sCD40L and 24 hours urine protein excretion, either UA or CRP( r =0. 591,0. 445,0. 539 ,all P ＜0. 01 ). No significant correlation was found between sCD40 L and systolic blood pressure, diastolic blood pressure,delivery gestational age, Cr, BUN, and platelet count( r =0. 178,0. 212, -0. 292,0. 144,0. 135, -0. 273,all P ＞0. 05). There was significant positive correlation between plasma sCD40 and sCD40L ( r =0. 707 ,P ＜0. 01 ). There was no relationship between the level of sCD40, sCD40L and the blood biochemical indexes in normotensive pregnant women ( P ＞ 0. 05 ). Conclusions The plasma concentrations of sCD40 and sCD40 L are significantly higher in pregnant women with preeclampsia compared with the control, which may be involved in the development of preeclampsia and contribute to the kidney damage. The variance levels of sCD40 and sCD40L may be also related to the severity of preeclampsia.
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Objective To investigate the correlation of the expressions of angiopoietin-2 (Ang-2) and angiopoietin-2 receptor(Tie-2)in serum and placenta with preeclampsia. Methods From May 2009 to April 2010, 62 women with preeclampsia who delivered in Affiliated Hospital of Qingdao University Medical College were recruited in the study, including 30 women with moderate preeclampsia (MPE group) and 32 women with severe preeclampsia (SPE group). Another 30 healthy pregnant women were taken as control group. ELISA was used to measure the serum Ang-2 in these women. Semiquantitative reverse transcription (RT)-PCR was used to investigate the expressions of Ang-2 mRNA and Tie-2 mRNA in placenta. Western blot was used to determine the expression of Ang-2 protein in placenta. Results (1) The serum concentrations of Ang-2 in MPE group and SPE group were (5.4 ± 1.8) μg/L and (5. 1 ± 1.7) μg/L,respectively. Both were significantly lower than that in control group (16. 2 ± 4. 5) μg/L (P<0. 01).There was no significant difference between MPE group and SPE group (P > 0. 05). (2) The expressions of Ang-2 mRNA in placenta of MPE group (2. 1 ± 0. 7) and SPE group (2. 0 ± 0. 6) were both significantly lower than that of control group (5.8 ± 0. 8; P<0. 01). But there was no significant difference in Ang-2 mRNA expression between MPE group and SPE group (P>0. 05). (3) No significant difference was found in the expressions of Tie-2 mRNA in placenta among MPE group (1. 33 ±0. 04), SPE group (1.35 ±0. 05) and control group (1.34 ± 0. 04; P > 0. 05). (4) The expressions of Ang-2 protein in placenta of MPE group (2.0 ± 0. 8) and SPE group (2. 0 ± 0. 8) were both significantly lower than that of control group (5.7 ±0. 9; P <0. 0l), while no significant difference was found between MPE group and SPE group (P >0. 05) . (5) In MPE group and SPE group, the serum concentrations of Ang-2 were positively correlated with the levels of Ang-2 mRNA and Ang-2 protein in placenta(r =0. 651, 0. 627; P <0. 01). Conclusions Decreased expressions of Ang-2 mRNA and Ang-2 protein in placenta reduced serum concentration of Ang-2. Low expression of Ang-2 may be involved in the pathophysiological process of preeclampsia by affecting the formation of placenta in early pregnancy.
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Objective To investigate the expression of macrophage migration inhibitory factor (MIF) and CD_(74), the receptor of MIF, in preeclamptic placenta and its correlation with the pathogenesis of preeclampsia.Methods From March 2008 to November 2008,69 preeclamptic women who delivered in the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College,were recruited,including 33 women with mild preeclampsia (MPE group) and 36 women with severe preeclampsia (SPE group).Another 43 healthy pregnant women were taken as control group.Immunoturbidimetry was applied to measure the concentrations of C-reactive protein (CRP) in maternal blood.The expressions of MIF and CD_(74) in placenta were tested with immunohistochemistry and the expressions of MIF mRNA and CD_(74) mRNA were detected by semiquantitative RT-PCR.The relationship between maternal blood level of CRP and MIF mRNA and CD_(74) mRNA in placenta was analyzed in the MPE and SPE group.Results (1) MIF and CD_(74) were expressed in the placenta of all pregnant women in the 3 groups, as shown in brown-yellow color, and significantly higher expression was found in the MPE and SPE group.(2) The expression of MIF mRNA and CD_(74) mRNA in the MPE group (0.70±0.13 and 0.96±0.16), SPE group (0.88 ± 0.12 and 1.08 ± 0.15) were significantly higher than in the control group (0.67 ± 0.11 and 0.83 ± 0.14) (P < 0.01), and statistical significance was also found between the MPE and SPE group (P <0.01).(3)The maternal blood concentrations of CRP in the MPE and SPE group were significantly higher than in the control group [(15.3±7.0) mg/L and (21.6±9.1)mg/L vs (4.8 ± 1.8) mg/L, P <0.01] , and significant difference was also found between the MPE and SPE group (P <0.01).(4) In the two preeclamptic groups, the blood concentrations of CRP were positively correlated with the expression of both MIF mRNA(r =0.67 ,P <0.01)and CD_(74) mRNA(r =0.83 ,P <0.01) in placenta.Positive correlation was also found between the levels of MIF mRNA and CD_(74) mRNA in placenta (r =0.93 ,P < 0.01).Conclusions Overexpression of MIF and CD_(74) in the placenta may up-regulate the CRP level in maternal blood, resulting in systemic inflammatory reaction and vascular endothelium damage which may be involved in the pathogenesis of preeclampsia.
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To study the level of macrophage migration inhibitory factor (MIF) in serum and the expression of MIF mRNA in abdominal subcutaneous adipose tissue,and to investigate its impact on insulin resistance and islet β-cell dysfunction in gestational diabetes mellitus (GDM).120 pregnancy women from the Affiliated Hospital of Qingdao University Medical College and Taian Central Hospital were enrolled,including 60 GDM women and 60 women with normal glucose tolerance (NGT).The serum MIF in GDM group was higher than that of NGT group [(3.58±1.02 vs 1.23±0.62) ng/ml,P<0.01].Multiple stepwise regression analysis showed that body mass index was an independent affective factor of the serum levels of MIF (r2 =0.516).The serum levels of MIF and the expressions of MIF mRNA in abdominal subcutaneous adipose tissue were significantly higher in GDM group than NGT group.MIF may contribute to insulin resistance and β-cell dysfunction in GDM.Body mass index seems to be an independent factor in affecting the serum levels of MIF.
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Objective To investigate the role of oxidized low-density lipoprotein (oxLDL) and lectin-like oxidized low-density lipoprotein receptor-1 (LOX-I) in pre-ecalmpsia. Methods From June 2007 to January 2008,73 women with pre-eelampsia who delivered in the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College,were recruited, including 35 women with mild pre-eclampsia (MPE group) and 38 women with severe pre-eclampsia(SPE group). And 45 healthy pregnant women were taken as control group. Enzyme-linked immunosorbent assay (ELISA) was used to measure the plasma concentrations of oxLDL in these women. Semiquantitative RT-PCR and Western blot were used to investigate the expression of LOX-1 mRNA and protein in placenta. The expression of caspase-3 mRNA in placenta was determined using Semiquantitative RT-PCR. Results (1) The plasma concentrations of oxLDL in MPE group ( 0.42 ± 0.11 ) mg/L, SPE group ( 0.68 ± 0.12 ) mg/L, were significantly higher compared with control group (0.35 ± 0.14 )mg/L( P < 0.01 ) and the concentrations of oxLDL in MPE group were significantly higher compared with SPE group (P<0.01 ). (2) The expression of LOX-1 mRNA in placenta of MPE group(0.70 ±0.10) and SPE group(0.84 ±0.08) were significantly higher than that in control group(0.58 ± 0.11 ) ( P<0.01 ) and the expression of LOX-1 mRNA in MPE group was significantly higher than that of SPE group (P<0.01 ). (3) The expression of LOX-1 protein in placenta of MPE group (0.79±0.15 )and SPE group(0.90±0.12) were significantly higher compared with control group(0.68 ±0.11)( P<0.01 ), while the expression of LOX-1 protein of MPE group was significantly higher compared with SPE group (P <0.01 ). (4) The expression of caspase-3 mRNA in placenta of MPE group(3.82± 0.18) and SPE group(5.39±0.14) were significantly higher than that in control group(2.19±0.20) (P <0.01 ), and the expression of caspase-3 mRNA in MPE group was significantly higher than that of SPE group (P<0.01 ). (5) In pre-eclampsia groups, the levels of LOX-1 mRNA in placenta were positively correlated with the plasma concentrations of ox LDL ( r=0.93, P<0.05 ), and caspase-3 mRNA expression were positively correlated with the expression of LOX-1 mRNA in placenta( r=0.84, P<0.05). Conclusion Increased levels of oxLDL in plasma may induce excess expression of LOX-1 in placenta, which may be involved in the pathophysiological processes of pre-eclampsia.
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Objective To investigate the expression of retinol binding protein 4(RBP-4) in maternal scrota and subcutaneous adipose tissue and its relationship with insulin resistance(IR)in gestational diabetes mellitus(GDM).Methods From May 2008 to April 2009,62 pregnant women who underwent elective cesarean section in the Department of Obstetrics.Affiliated Hospital of Qingdao University Medical College,were recruited,including 32 with GDM(GDM group)and 30 with normal glucose tolerance test(control group).Enzyme-linked immunosorbent assay(ELISA)was used to determine the serum concentrations of RBP-4 and radio immunoassay to measure the serum levels of fasting insulin (FINS).Fasting plasma glucose (FPG) was tested by glucose oxidase,and the Home model insulin resistance index(HOMA-IR)Was caleulated.Reverse transcription PCR(RT-PCR)and Western blot were applied to investigate the expression of RBP-4 mRNA and protein in subcutaneous adipose tissue.The correlations between the expression of RBP4 mRNA and protein in subcutaneous adipose tissue and the serum RBP-4 concentrations and HOMA-IR were analyzed. Results (1) The serum concentrations of RBP-4、FINS、FPG、HOMA-IR in GDM group were significantly higher compared with the control group [(27. 0 ±1.2) mg/L vs. (19.4±1.8)mg/L, (12.1±1.4)mU/L vs. (8.3±0.8)mU/L, (5.3±0.9)mmoL/L vs.(4. 1±0. 6) mmol/L, 2. 5 ± 0. 2 vs. 1.5 ± 0. 1, P < 0. 05, respectively]. ( 2 ) The expression of RBP-4 mRNA and protein in subcutaneous adipose tissue in the GDM group were significantly higher than that of the control group (0. 76 ± 0. 12 vs. 0. 53 ± 0. 06, 0. 74± 0. 09 vs 0. 54 ± 0. 06, P < 0. 05). (3) In the GDM group, the expression of both RBP-4 mRNA and protein in the subcutaneous adipose tissue were positively correlated with HOMA-IR(r=0. 575 and 0. 851, P < 0. 05). The serum concentration of RBP-4 were also positively correlated with HOMA-IR (r = 0. 635, P < 0. 05 ). No correlations was found between the expressions of RBP-4 mRNA and protein in subcutaneous adipose tissue with the serume RBP-4 concentrations. Conclusion High expression of RBP-4 mRNA in subcutaneous adipose tissue and the elevation of serum RBP-4 levels in GDM women may contribute to IR.
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Objective To investigate the expression and significance of Rho-associated protein kinaseⅡ(Rock Ⅱ)in preeclamptic placenta and umbilical artery.Methods Semiquanfitative RT-PCR and Western blot were used to investigate the expression of RockⅡmRNA and RockⅡprotein in placenta and umbilical artery from 35 women with moderate preeclampsia(MPE group)、38 women with severepreeclampsia(SPE group)and 45 normal third trimester pregnant women(control group),the S/D value of umbilical artery was examined by ultrasound.Results (1)The expression of Rock Ⅱ mRNA of Dlacenta in MPE group(0.82±0.14)and SPE group(0.93±0.13)were signifieantly higher than that in control group (0.70 ±0.12,P＜0.01).The expression of Rock Ⅱ protein of placenta in MPE group(0.79±0.15)and SPE group(0.92±0.12)were significantly higher compared with control group(0.68±0.11,P<0.01).The expression of Rock Ⅱ mRNA and protein of placenta in SPE group were higher compared with MPE group(P＜0.01).(2)The expression of Rock Ⅱ mRNA of umbilical artery in MPE group(0.69±0.13)and SPE group(0.55±0.12)were significantly lower than that in control gmup(0.76±0.10,P＜0.01).The expression of RockⅡ protein of umbilical artery in MPE group(0.68±0.10)and SPE group(0.51±0.12)were lower compared with control group(0.75±0.13,P＜0.01).The expression of RockⅡ mRNA and protein of umbilical artery in SPE group were significantly lower compared with MPE group(P＜0.01).(3)There were no correlations between the expression of RockⅡ mRNA and protein in placenta and umbilical artery and the S/D value and birth weight(P＞0.05).Conclusion The upregulated expression of Hock Ⅱ in placentas and downregulated expression in umbilical artery may be a compensation in preeclampsia.
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Objective To investigate the expression and significance of serum soluble fms-like tyrosine kinase 1 ( sFlt-1 ) in preeclampsia placenta. Methods Immunohistochemistry and semi-quantitative RT-PCR were used to investigate the expression of sFlt-1 and sFlt-1 mRNA in placenta of 30 women with preeclampsia (including mild and moderate preeclampsia 11 cases, severe preeclampsia 19 cases) and 45 normal pregnant women (including first trimester pregnancy 18 cases, 2nd trimester pregnancy 12 cases, and 3rd trimester pregnancy 15 cases). The levels of vascular endothelial growth factor(VEGF) and sFlt-1 in serum were detected by enzyme linked immunosorbent assay. Results (1) The expression of sFlt-1 mRNA was significantly higher in preeclampsia group (0. 90?0. 11) compared with the third trimester group (0. 80?0. 06; P
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Objective To study the blood pressure changing patterns in normal pregnancy and pregnancy induced hypertension(PIH). Methods 24 hour ambulatory blood pressure monitoring (ABPM) were performed in 38 cases of normal pregnancy and 36 cases of PIH during 18～20,28～30 and 38～40 gestational week. Results The 24 hour mean blood pressure were increased with pregnancy advanced. In normal pregnancy, a typical circadian rhythm of blood pressure pattern was shown that it increases during the day and decreased during the night with the disparity of (16.3? 4.1)%, (14.1?3.2)% and (15.7?2.0)% in three different gestational week respectively. In moderate and severe PIH, the 24 hour mean blood pressure,blood pressure load and the frequence of abnormal rhythm were significantly higher than those in mild PIH and normal pregnancy, while the disparity between day and night blood pressure was lower. Conclusion The blood pressure patterns show a typical circadian rhythm in most normal pregnancy. The more severe the PIH is, the higher rate of abnormal blood pressure rhythm occured.