Preeclampsia (PE), new onset hypertension during pregnancy, affects 5‐7% of all pregnancies in the U.S. and is associated with reduced fetal weight, increased inflammation, vascular endothelial dysfunction (increased endothelin‐1 (ET‐1) and decreased nitric oxide (NO)) and hypertension. To date the best treatment remains early delivery of the feto‐placental unit. Activated lymphocytes during normal pregnancy (NP) express progesterone receptors, which stimulate a protein called Progesterone Induced Blocking Factor (PIBF) that is reduces during hypertensive pregnancy disorders. Therefore, this study was designed to test the hypothesis that progesterone, in the form of 17‐hydroxyprogesterone caproate (17‐OHPC), reduces inflammation, markers of endothelial dysfunction while reducing blood pressure and prolong time to delivery in PE women. In our ongoing clinical trial, PE participants received 17‐OHPC (250 mg, I.M.) with blood draws before and after injection. Placentas were collected at delivery. PIBF was 18.6 +/‐1.0 pg/mL in NT (n=4), 14.53 +/‐ 1.0 pg/mL in PE (n=10, p<0.05), and 15.78 +/‐0.85 in PE+17‐OHPC (n=6). Placental CD4+ T cells were 6.5+/‐ 2.7 in PE (n=3), 4.6+/‐ 2.0 in PE +17‐OHPC (n=4). Circulating CD4+ T cells were 18.23 +/‐4.8 in PE (n=4), 14.4+/‐1.2 % gate in PE+17‐OHPC (n=5). Placental TH2 cells were 81.7+/‐10.6% gate in PE and 84.7+/‐11.0 in PE+17‐OHPC. Placental and circulating NK cells were 20.84 +/‐ 6.8 % gate, 7.2 +/‐ 2.0 in PE which reduced to 4.7+/‐1.03, 5.3 +/‐ 1.5 % gate in PE+17‐OHPC. Circulating TNF‐alpha was 32.0 +/‐ 3.4 pg/mL in PE (n=8), which decreased to 21.1+/‐5.5 in PE+17OHPC (n=4). Circulating ET‐1 was 2.53+/‐ 0.4 pg/mL in healthy normal pregnant (NP, n=5), 6.7 +/‐ 1.4 in PE (n=18, p<0.05) and 4.9 +/‐ 1.3 in PE+17‐OHPC. Placenta preproendothelin ‐1 (PPET‐1) increased 1.5 fold change in PE (n=4) compared to NP, which was reduced to 0.96 in PE+17‐OHPC (n=4). Importantly, endotheling‐1 measured in HUVECS media treated with PE sera was 68 +/‐ 22 pg/mg of protein in PE which reduced to 57 +/‐19 in PE+17OHPC (n=6). Moreover, circulation nitrate‐nitrite was 50 +/‐ 9 uM in PE and significantly increased to 94 +/‐14 in PE+17‐OHPC. 17‐OHPC prolonged time of delivery beyond 72h on average and average systolic blood pressure was 151 +/‐ 5 mmHg in PE (n=18) and 137+/‐4 in PE+17‐OHPC (n=13). Our results suggest that 17‐OHPC reduced inflammation, markers of endothelial dysfunction, lowered blood pressure and prolonged time do delivery in PE women.
Placenta ischemia, the initiating factor in preeclampsia (PE), is associated with intrauterine growth restriction (IUGR) and increased blood pressure (BP) in offspring. Yet, the only treatment for PE is delivery of the baby and placenta. The Reduced Uterine Perfusion Pressure (RUPP) rat model induced by placental ischemia at gestational day 14 (G14) mimics many facets of human PE including pregnancy-specific hypertension, an increase in the agonistic ANG II Type 1 receptor autoantibody (AT1-AA), IUGR and increased BP in the offspring. Inhibition of AT1-AA using an epitope-binding inhibitory peptide ('n7AAc') attenuates increased BP at gestational day 19 in the RUPP. Yet, whether use of ‘n7aac’ improves fetal growth and mitigates increased BP in the offspring is unknown. Thus, we tested the hypothesis that maternal administration of ‘n7aac’ improves fetal growth by attenuating reduced uterine blood flow and impaired placental remodeling. Sham or RUPP surgery was performed at G14 with administration of vehicle or ‘n7aac’ (144μg/day) via mini osmotic pump until gestational day 20 (G20). At G20 uterine artery resistance index was significantly elevated in vehicle RUPP (0.69±0.02 mm/s n=10) compared to vehicle Sham (0.48±0.02 mm/s n=8) (P<0.0001) and not increased in treated RUPP (0.49±0.02 mm/s n=10) or treated Sham (0.48±0.02 mm/s n=9). Fetal weight was significantly reduced in vehicle RUPP (3.24±0.2 g) compared to vehicle Sham (3.92±0.05 g) (P=0.013) and not decreased in treated RUPP (3.70±0.04 g) or Sham (3.98±0.10 g). Litter size of viable pups at G20 was only reduced in treated RUPP (5.3±1.4) compared to vehicle Sham (11.56±0.7) (P=0.003). Importantly, using in vivo imaging, little to no auto fluorescence of rhodamine-labeled peptide (480 μg/kg/day, n=4) was detectable in the pups at G20. Thus, our results demonstrate that maternal treatment with ‘n7aac’ in the RUPP rat model of PE improve UARI, which is associated with improved fetal weight at G20 in response to placental ischemia. Whether this benefit continues to birth and mitigates increased BP in IUGR offspring is unknown but is the focus of future studies. In conclusion, inhibition of the AT1AA during PE may not only provide benefit to the mother, but may also be associated with benefit in the offspring.
Preeclampsia (PE), new onset hypertension during pregnancy, is the leading cause of death and morbidity for the mother and low birth weight in offspring. PE women have activated B cells producing agonistic autoantibodies to the angiotensin II type I receptor (AT1-AA). We have shown Rituximab (R), used clinically for B cell depletion, lowers mean arterial pressure (MAP), B cells, and AT1-AA in the RUPP rat model of PE. Clinical studies show no untoward effects on offspring of pregnant women maintained on R for treating lymphoma, however, R is not used during PE therefore, effects of maternal B cell depletion on offspring survival and growth in response to placental ischemia is unknown. We hypothesize that R will deplete maternal B cells in RUPP rats without worsening the effect of placental ischemia on pup growth and survival. To test this hypothesis, R (250 mcg/kg) was given on gestation day (GD) 14 via mini-osmotic pump. On GD 19 B cells were measured by flow cytometry, and MAP and pup weights were recorded. A separate group of dams were allowed to deliver, pup weights were recorded within 12 hours and weekly until 16 weeks, and B cells were analyzed. A one-way ANOVA was used for statistical analysis. MAP increased in RUPP 123±2 (n=19, p<0.05) compared to NP controls 101±1 (n=18) and was 106±3 mmHg in RUPP+R (n=8, p<0.05). On GD19, maternal circulating B cells were 16±2 % (n=14) in RUPPs, 8±2 % in NP rats, (n=7, p<0.05), and 5.5±1% gate in RUPP+R (n=5, p<0.05). RUPP male and female offspring were smaller 5.11±0.23 g, 5.19±0.14 g (n=4, n=4) at birth than NP offspring 6.09±0.15 g, 5.87±0.12 g (n=6, p<0.05; n=6, p<0.05) or RUPP+R offspring 5.75±0.24 g, 5.36±0.28 g (n=6, p=0.11; n=6, p=0.67). At 12 weeks, male and female RUPP offspring had elevated circulating B cells (21±3, 20±1 % (n=6; n=9)) compared to NP (1±0.23, 1.6±0.06 % (n=4, p<0.05; n=3, p<0.04)) which was normalized in RUPP+R offspring (0.4±0.1, 0.3±0.03 % gate (n=3, p<0.05; n=8, p<0.05)). At 16 weeks, B cells were comparable in male offspring from NP (0.78±0.09 % (n=10)) and RUPP+R rats (0.80±0.04 % gate (n=3)). Our findings indicate that R lowers maternal circulating B cells and MAP in RUPP rats and improves fetal weight and circulating B cells, indicating that R does not worsen adverse fetal outcomes in response to placental ischemia.
Preeclampsia (PE), new onset hypertension during pregnancy, is the leading cause of death and morbidity world‐wide for the mother and fetus. Women with PE have elevated inflammatory cytokines, cytolytic natural killer (cNK) cells, and B cells producing agonistic antibodies to the angiotensin II type I receptor (AT1‐AA). The Reduced Uterine Perfusion Pressure Rat Model of PE (RUPP) exhibits many characteristics of PE. We have shown Rituximab, a chimeric monoclonal antibody used clinically for B cell depletion, lowers blood pressure, AT1‐AA, and cytokines in RUPP rats. However, we don't know the effect of maternal B cell depletion on placental cNK cells, or offspring survival and growth. Moreover, recent studies have touted that Rituximab doesn't work in rodents nor are B cells important for hypertension in RUPP rats. Therefore, we hypothesize that Rituximab will deplete circulating B cells and lower AT1‐AA stimulated cNK cells and blood pressure in response to placental ischemia in RUPP rats. Rituximab (250 mcg/kg) was given on gestation day (GD) 14 via mini‐osmotic pump. Blood and tissues were collected, blood pressure (MAP), pup weight, and NK cells were measured by flow cytometry in the blood and placenta on GD 19. A separate group of dams were allowed to deliver and birth weight of pups were recorded within 12 hours. Weights were measured weekly until 12 weeks. A one‐way ANOVA was used for statistical analysis. On GD19, circulating B cells were 15.67±2.28 % gate (n=14) in RUPP rats which was elevated compared to NP rats, 7.04±2.3 % gate (n=6, p<0.05), and was normalized with Rituximab (4.21±0.61 % gate (n=3, p<0.05)). MAP increased in RUPP 123±2 mmHg (n=19, p<0.05) compared to NP controls 101±1 mmHg (n=18) and was normalized with Rituximab 106±3 mmHg (n=8, p<0.05). Circulating and placental cNK cells were 0.22±0.14, 0.64±0.48 % gate in NP rats (n=9, n=6), 1.36±0.32, 1.83±0.49 % gate in RUPP rats (n=16 p<0.05, n=10), which was decreased to 0.01±0.01, 0.81±0.50 % gate with Rituximab (n=5, p<0.05, n=5). RUPP pup weight, 1.78±0.06 g (n=19), was reduced compared to NP pup weight, 2.29±0.14 g (n=18, p<0.05) and was unchanged by Rituximab, 1.82±0.16 g (n=7). RUPP and Rituximab male and female offspring were smaller at birth than NP offspring. Although there no differences among the female offspring, RUPP and Rituximab male offspring remained smaller at 12 weeks of age compared to NP offspring. Our findings indicate that Rituximab indeed lowers maternal circulating B cells, cNK cells and blood pressure in response to placental ischemia in pregnant dams and may not exacerbate adverse fetal outcomes in response to placental ischemia.
Preeclampsia (PE), new onset hypertension during pregnancy, is the leading cause of death and morbidity for the mother and low birth weight in offspring. Low birth weight has been shown to cause offspring to be at high risk for cardiovascular and metabolic disorders later in life. PE has been associated with chronic immune activation including T helper cells and natural killer cells. PE women also have activated B cells producing agonistic autoantibodies to the angiotensin II type I receptor (AT1‐AA). AT1‐AA has been implicated in numerous pathways in the pathophysiology of PE. We have shown Rituximab (R), used clinically for B cell depletion, lowers mean arterial pressure (MAP), B cells, and AT1‐AA in the reduced uterine perfusion pressure (RUPP) rat model of PE. Clinical studies show no untoward effects on offspring of pregnant women maintained on R for treating lymphoma. R is not used during PE, therefore, effects of maternal B cell depletion on offspring survival and growth in response to placental ischemia is unknown. We hypothesize that R will deplete maternal B cells in RUPP rats without worsening the effect of placental ischemia on pup growth and survival. To test this hypothesis, the RUPP procedure was performed and R (250 mcg/kg) was given on gestation day (GD) 14 via mini‐osmotic pump. On GD 19, B cells were measured by flow cytometry, and MAP and pup weights were recorded. A separate group of dams were allowed to deliver, pup weights were recorded within 12 hours of birth and weekly until 16 weeks, and B cells were analyzed. A one‐way or two‐way ANOVA was used for statistical analysis. MAP increased in RUPP 123±2 (n=19, p<0.05) compared to NP controls 101±1 (n=18) and was 106±3 mmHg in RUPP+R (n=8, p<0.05). On GD19, maternal circulating B cells were 16±2 % (n=14) in RUPPs, 8±2 % in NP rats, (n=7, p<0.05), and 5.5±1% gate in RUPP+R (n=5, p<0.05). RUPP male and female offspring tended to be smaller (5.11±0.23 g, 5.19±0.14 g; n=4, n=4) at birth than NP offspring (6.09±0.15 g, 5.87±0.12 g; n=6, p<0.064; n=6, p<0.948) or RUPP+R offspring (5.75±0.24 g, 5.36±0.28 g; n=6, p=0.291; n=6, p>0.999). At 12 weeks, male and female RUPP offspring had elevated circulating B cells (21.24±2.92, 20.04±0.72 %; n=6; n=9) compared to NP (1.38±0.23, 1.63±0.06 %; n=4, p<0.05; n=3, p<0.04) which was normalized in RUPP+R offspring (0.40±0.10, 0.27±0.03 % gate; n=3, p<0.05; n=8, p<0.05). At 16 weeks, B cells were comparable in male and female offspring from NP (0.78±0.09 %, n=10; 1.06±0.21 % gate, n=6) and RUPP+R rats (1.21±0.34% gate, n=5; 1.62±0.34% gate, n=5), but were elevated in RUPP male and female offspring (2.45±0.27% gate, n=9; 1.68±0.45% gate, n=13) compared to NP (p<0.05; p>0.99) and RUPP+R (p<0.05; p>0.99). Our findings indicate that R lowers maternal circulating B cells and MAP in RUPP rats and improves fetal weight and circulating B cells, indicating that R does not worsen adverse fetal outcomes in response to placental ischemia.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.