Supplementing punicalagin reduces oxidative stress markers and restores angiogenic balance in a rat model of pregnancy-induced hypertension

Pre-eclampsia (PE) is a pregnancy disorder that is characterised by severe hypertension and increased risks of foetal and maternal mortality. The aetiology of PE not completely understood; however, maternal nutrition and oxidative stress play important roles in the development of hypertension. The treatment options for PE are currently limited to anti-hypertensive drugs. Punicalagin, a polyphenol present in pomegranate juice, has a range of bioactive properties. The effects of supplementation with punicalagin on angiogenesis and oxidative stress in pregnant rats with induced hypertension were investigated. The pregnant rats were randomly divided into five experimental groups (n=12 per group). Hypertension was induced using an oral dose of NG-nitro-L-arginine methyl ester (L-NAME, 50 mg/kg/day) on days 14–19 of pregnancy. Punicalagin (25, 50 or 100 mg/kg) was given orally on days 14–21 of pregnancy. Punicalagin treatment at the tested doses significantly reduced diastolic, systolic, and mean arterial blood pressure in L-NAME treated rats from day 14. Punicalagin also restored angiogenic balance by increasing the expression of vascular endothelial growth factor and downregulating vascular endothelial growth factor receptor-1/fms-like tyrosine kinase-1. Punicalagin, significantly increased the placental nitric oxide levels as compared to PE group. The increased levels of oxidative stress in rats with PE were markedly decreased by treatment with punicalagin. Punicalagin at the tested doses markedly (p < 0.05) enhanced the placental antioxidant capacity in L-NAME-treated rats. The raised catalase activity observed following L-NAME induction was significantly (p < 0.05) and restored to normal activity levels in punicalagin treatment. Further, 100 mg dose of punicalagin exhibited higher protective effects as compared to lower doses of 25 and 50 mg. This study shows that supplementation with punicalagin decreased blood pressure and oxidative stress and restored angiogenic balance in pregnant rats with induced PE.

Effects of histologic chorioamnionitis induced by lipopolysaccharide intra-amniotic injection on fetal lung maturation and development in preterm fetal rats / 中华围产医学杂志

Objective With various doses of lipopolysaccharide (LPS) intra-amniotic injection in pregnant rats to establish histologic chorioamnionitis (HCA) model,and to determine the effects of HCA on fetal lung maturation and development in preterm fetal rats.Methods Thirty pregnant Sprague-Dawley rats were randomly assigned to four groups.On 19 days of gestation,rats in three model groups (n=8) were given intra-amniotic injection of LPS 2,8,10 μg,respectively,and those in the only one control group (n=6)was given normal saline (NS) 40 μl.Fetal rats were taken out 48 hours after LPS injection.Placenta,fetal membrane,fetal lung and umbilical cord were collected for pathological examination.mRNAs of surfactant protein (SP)-A,B and C and keratinocyte growth factor (KGF) in lung were determined by quantitative real-time polymerase chain preaction.KGF protein expression in lung was measured by immunohistochemistry.Morphologic observation of lung was performed on postnatal day 3.R× C table Chi-square test,KruskalWallis test and analysis of variance were used as statistical methods.Results (1) Fetal rat mortality in control,LPS2,LPS8 and LPS10 group was 0.0％ (0/55),22.5％ (18/80),51.4％ (36/70) and 87.5％(35/40),respectively,which increased with increasing dose of LPS (x2=46.183,P＜0.005).(2) HCA,fetal lung inflammation and umbilical vasculitis were induced in LPS groups,and the severity was dosedependent (fetal lung inflammation:Hc=39.84,HCA:Hc=41.13,umbilical vasculitis:Hc=41.52,all P＜0.01).(3) The expression of SP-A,SP-B,SP-C and KGF mRNAs in the four groups (control,LPS2,8,10) had statistical differences (SP-A mRNA:0.99±0.28,2.48±0.34,1.09±0.31 and 0.09±0.09,F=78.051; SP-B mRNA:0.99±0.34,2.23±0.53,1.49±0.51 and 0.14±0.06,F=28.327; SP-C mRNA:1.20±0.39,2.00±0.20,1.04±0.37 and 0.12±0.16,F=39.546; KGF mRNA:0.97±0.19,2.18±0.61,0.93±0.09 and 0.21±0.11,F=37.544; all P＜0.01).All mRNA expressions in LPS2 group were higher than those in the control group and those in LPS10 group was lower (all P＜0.05).(4) The expression of KGF protein in LPS2 group was significantly higher than that in other groups (0.60±0.20 vs 0.28±0.12,0.37±0.22,0.24±0.12,F=17.280,all P＜0.01).(5) Alveolarization was significantly inhibited in LPS8 group on postnatal day 3,and less maturity of pulmonary tissue was observed.Conclusions Various doses of LPS intra-amniotic injection in rats could induce HCA,fetal lung inflammation and umbilical vasculitis with different degrees.Histological inflammation would be worse with increasing LPS dosage.Moderate inflammation could promote lung maturation without inducing bronchopulmonary dysplasia,and KGF may play a role in this process.

Expression of KLF-8 and MMP-9 in placentas and their relationship with the pathogenesis of preeclampsia / 中华妇产科杂志

Objective To evaluate the expression of Krüppel-like factor 8 (KLF-8) and matrix metalloproteinase 9 (MMP-9) in placentas and their relationship with the pathogenesis of preeclampsia (PE).Methods Twenty-two women with PE (mild PE:4 cases; severe PE:18 cases) who received cesarean sections in the First Affiliated Hospital of Chongqing Medical University from September 2011 to March 2012 were recruited as the PE group (n =22).And twenty women who received elective term cesarean section without perinatal complications were chosen as the control group (n =20).Placentas were collected and immunohistochemical SP method were employed to detect the localization of KLF-8 protein.KLF-8 mRNA level was determined by quantitative real-time PCR technique and western blot analysis was used to quantify KLF-8 and MMP-9 protein levels.Results (1) There was no difference of KLF-8 protein distribution in placentas of the PE group and the control group.It was mainly located in the nuclear and cytoplasm of syncytiotrophoblasts.KLF-8 immunostaining was apparently decreased in the placentas of preeclamptic women when compared with the control group.(2)The KLF-8 mRNA levels were significantly decreased in placentas of the PE group (0.69 ±0.08) compared to those of the control group (1.14 ±0.09,P ＜0.01).(3) KLF-8 and MMP-9 protein levels significantly decreased in the PE placentas (0.68 ±0.05 and 0.21 ± 0.03) when compared to the control group (0.94 ± 0.06 and 0.34 ± 0.03,respectively,P ＜ 0.01).(4) There was a positive correlation between the expression of KLF-8 and MMP-9 protein in the placentas from PE and normal pregnancies (r =0.64,P ＜ 0.01).Conclusions KLF-8 mRNA and protein levels were decreased in placentas of PE patients compared to those of normotensive women.KLF-8 protein was primarily located in the invasion-related trophoblast cells and its expression had a positive correlation with MMP-9 levels.KLF-8 might have an important role in the pathogenesis of PE by regulation of trophoblast invasion.