Alaa Amash

Placental Secretion of Interleukin-1 and Interleukin-1 Receptor Antagonist in Preeclampsia: Effect of Magnesium Sulfate

Preeclampsia is a pregnancy-specific disorder characterized by hypertension and systemic endothelial dysfunction. Interleukin (IL)-1β is a possible mediator of maternal endothelial dysfunction in preeclampsia. Serum IL-1β as well as its natural inhibitor IL-1 receptor antagonist (IL-1Ra) were reported to be increased in women with preeclampsia. In the current study, we addressed the role of the placenta in controlling the circulatory levels of IL-1β and its natural inhibitor IL-1Ra in preeclampsia, and the possible effect of magnesium sulfate (MgSO(4)) on these levels. Using an ex vivo placental perfusion system, placentas from preeclamptic (n = 9) and normotensive (n = 6) pregnancies were perfused in presence or absence of MgSO(4). Perfusate samples were collected from the maternal and the fetal circulations of the perfusion system, and IL-1β and IL-1Ra were examined by enzyme-linked immunoassay (ELISA). Preeclamptic placentas secreted higher levels of IL-1β (P < 0.001), and a tendentious higher levels of IL-1Ra, mainly into the maternal circulation, as compared with normotensive placentas, although no differences in IL-1β:IL-1Ra ratio were detected. However, there was only tendentious increase in the secretion levels of IL-1β or IL-1Ra into the fetal circulation of preeclamptic placentas, when compared with normotensive placentas. Administration of MgSO(4) to preeclamptic placentas resulted in an attenuation of the increased secretion of IL-1β into the maternal circulation (P < 0.001), and in a tendentious reduction in IL-1Ra. However, IL-1β:IL-1Ra ratio in preeclamptic placentas was not affected by MgSO(4). Interestingly, exposure of normotensive placenta to MgSO(4) resulted only in increased levels of IL-1Ra in the maternal circulation, without affecting IL-1β levels or IL-1β:IL-1Ra ratio. These findings suggest that the placenta may contribute to the elevation in serum IL-1β and IL-1Ra in preeclampsia by increased secretion of these cytokines into the maternal circulation, and that MgSO(4) is able to attenuate this increased secretion of IL-1β, and possibly IL-1Ra, in preeclampsia.

Magnesium Sulfate Normalizes Placental Interleukin-6 Secretion in Preeclampsia

Interleukin-6 (IL-6) is one of the main proinflammatory mediators of hypertension and endothelial dysfunction in preeclampsia. In this study, we investigated the capacity of the preeclamptic placenta to secrete IL-6 and the effect of magnesium sulfate (MgSO(4)) on it. Placentas from normotensive (37-40 weeks) and preeclamptic (36-40 weeks) pregnancies were dually perfused for 6 h in the absence [normotensive (n = 3); preeclamptic (n = 4)] and presence [normotensive (n = 3); preeclamptic (n = 4)] of MgSO(4). Perfusate samples from the maternal and the fetal circulations were collected at each 30 min throughout the perfusion period and examined for IL-6 by enzyme-linked immunoassay. Statistical analysis was performed using the 2-way analysis of variance. In the absence of MgSO(4), IL-6 levels in the maternal and the fetal circulations of preeclamptic placentas (4.2 ± 1.3 and 0.9 ± 0.5 pg/mL/g cotyledon; respectively) were significantly higher, when compared with normotensive placentas (1.9 ± 0.5 and 0.2 ± 0.2 pg/mL/g cotyledon; respectively) (P < 0.05). Addition of MgSO(4) to the perfusate of normotensive placentas did not affect IL-6 secretion. However, exposure of preeclamptic placentas to MgSO(4) resulted in decreased IL-6 levels in the maternal circulations (1.7 ± 0.3 pg/mL/g cotyledon), when compared with the control group (P < 0.05). In the fetal circulation, the addition of MgSO(4) resulted only in a nonstatistical significant tendency toward decreased IL-6 levels, when compared with the control group. Our findings indicate that the perfused preeclamptic placenta secretes increased levels of IL-6 into the fetal and the maternal circulations and that MgSO(4) may normalize these increased secreted IL-6 levels.

Perfusion with lipopolysaccharide differently affects the secretion of interleukin-1 beta and interleukin-1 receptor antagonist by term and preterm human placentae

The aim of the present study was to examine the effect of lipopolysaccharide (LPS) on the secretion of the pro-inflammatory cytokine interleukin-1beta (IL-1beta) and of its natural inhibitor interleukin-1 receptor antagonist (IL-1Ra), by perfused human term and preterm placental tissue. Eight term and eight preterm placentae were collected immediately after delivery; four term and four preterm placentae were perfused with control medium (without LPS) and the other four term and four preterm placentae were perfused with medium containing LPS. The release of IL-1beta into the maternal compartment by term placenta was significantly higher than the release by preterm placenta (p<0.001). However, there were no significant differences between IL-1beta levels released into the fetal compartments of term and preterm placentae. No significant differences were observed in the release of IL-1Ra into the maternal and fetal compartments of term placenta, when compared to preterm placenta. Exposure to LPS significantly decreased the capacity of term placenta to release IL-1beta into the maternal compartment (p<0.001) and increased the capacity of term placenta to release IL-1Ra into the maternal and fetal compartments (p<0.001 and p=0.017, respectively). However, the capacity of preterm placentae to release IL-1beta and IL-Ra into the maternal and fetal compartments was not affected by LPS. IL-1beta was expressed by both term and preterm placentae before and after perfusion (+/- LPS), by epithelial cells of the amnion, chorion, by syncytiotrophoblast and stromal cells of villous tissue and by the decidua. IL-1Ra in term and preterm placentae was expressed before perfusion mainly in epithelial cells of the amnion. After perfusion of term placentae (+/- LPS), additional IL-1Ra expression was seen in epithelial cells of the amnion and in syncytiotrophoblast and stromal cells of villous tissue and by the decidua. However, perfusion of preterm placentae (+/- LPS) did not affect IL-1Ra expression. The localization of IL-1beta and IL-1Ra in both term and preterm human placental tissue suggests a their physiologic role. The data presented indicates that the IL-1 system in term and preterm placentae seems to be differently affected by LPS. Down-regulation in the release of the pro-inflammatory cytokine IL-1beta and the up-regulation of its antagonist (IL-1Ra) may be a part of the inflammatory response to infection in human term, but not preterm, placentae. The IL-1 system in term and preterm placentae seems to be differently affected by LPS.

Possible therapeutic effect of magnesium sulfate in pre-eclampsia by the down-regulation of placental tumor necrosis factor-alpha secretion

OBJECTIVE To examine the effect of magnesium sulfate (MgSO(4)) on tumor necrosis factor-alpha (TNF-alpha) secretion by preeclamptic placentas. STUDY DESIGN Cotyledons of six, term, normotensive and ten, pre-eclamptic placentas were dually perfused for six hours (6h), with MgSO(4) (6-7 mg %) in the maternal reservoir [normotensive (n = 3); pre-eclamptic (n = 5)], and with control medium (without MgSO(4)) [normotensive (n = 3); pre-eclamptic (n = 5)]. Perfusate samples from the maternal and the fetal circulations were collected every 30 min throughout the 6h of perfusion, and examined for TNF- alpha levels using ELISA. Statistical significance was determined using a 2-way analysis of variance. RESULTS Pre-eclamptic placentas perfused with control medium (without MgSO(4)) secreted higher levels of TNF-alpha into the fetal and the maternal circulations (1.60 +/- 0.59 pg/mL/g of cotyledon and 14.28 +/- 2.69 pg/mL/g of cotyledon, respectively), as compared to the fetal and maternal circulations of normotensive placentas (0.25 +/- 0.09 pg/mL/g of cotyledon and 6.73 +/- 1.11 pg/mL/g of cotyledon, respectively) (p < 0.01). Addition of MgSO(4) to normotensive placentas did not affect TNF-alpha levels in the fetal or maternal circulations. However, exposure of pre-eclamptic placentas to MgSO(4) significantly decreased TNF-alpha levels in both the fetal (0.89 +/- 0.09 pg/mL/g of cotyledon versus 1.6 +/- 0.59 pg/mL/g of cotyledon; p < 0.05) and the maternal circulations (4.74 +/- 2.78 pg/mL/g of cotyledon versus 14.28 +/- 2.69 pg/mL/g of cotyledon; p < 0.01). CONCLUSION Down-regulation of placental TNF-alpha secretion by MgSO(4) in pre-eclampsia might indicate a possible therapeutic effect for this agent in reducing maternal, endothelial dysfunction and in improving neonatal outcome in pre-eclampsia, by reducing TNF-alpha levels in maternal and fetal circulations.

Perfusion with magnesium sulfate increases sFlt-1 secretion only in the fetal side of placenta of women with preeclampsia

Objective: To examine the effect of magnesium sulfate (MgSO4) on sFlt (soluble fms-like tyrosine kinase)-1 in the fetal and maternal compartments of normotensive and preeclamptic placentas. Methods: Cotyledons of term normotensive and preeclamptic placentas were dually perfused for six hours, with control medium and MgSO4 (6–7 mg %) in the maternal reservoir. Perfusate sFlt-1 concentrations were measured. Results: Median sFlt-1 concentration was higher in the maternal than in the fetal side in both groups and perfusion media (p < 0.0001). When perfused with control medium, the maternal side median sFlt-1 concentration was higher in the preeclampsia than in the control group (p < 0.0001). After pefusion with MgSO4, the median maternal and fetal sides perfusate sFlt-1 concentration were higher in the preeclampsia than in the control group (p < 0.0001). In comparison to perfusion with control medium, the median sFlt-1 concentration of normal pregnant women decreased in the fetal and increased in the maternal side. In the preeclampsia group, only median fetal side sFlt-1 concentration increased. Conclusion: In contrast to normal pregnant women, perfusion with MgSO4 of preeclamptic placentas did not increase their sFlt-1 concentration. This may indicate that MgSO4 role may be limited to its anti-eclamptic and does not affect the anti-angiogenic state associated with preeclampsia.

The effects of magnesium sulfate on placental vascular endothelial growth factor expression in preeclampsia

Objective: To evaluate the effect of magnesium sulfate (MgSO4) on placental expression levels of vascular endothelial growth factor (VEGF). Materials and methods: Cotyledons of term normotensive and preeclamptic placentas were dually perfused for 6 h, with MgSO4 (6–7 mg%) in the maternal reservoir [normotensive (n = 3); preeclamptic (n = 4)] and with the control medium (without MgSO4) [normotensive (n = 3); preeclamptic (n = 6)]. After perfusion, placental tissue samples were collected from four different placental compartments (amnion, chorion, placental villous and decidua). The collected placental tissues were homogenized and examined for VEGF by ELISA. Statistical significance was determined using a two-way analysis of variance. Results: After perfusion with control medium, significantly lower levels of VEGF were detected in the chorion and placental villous compartments of preeclamptic placentas (70 ± 24 pg/g protein and 29 ± 11 pg/g protein; respectively), as compared with normotensive placentas (172 ± 80 pg/g protein and 51 ± 17 pg/g protein; respectively; p < 0.05). Exposure of preeclamptic placentas to MgSO4 resulted in decreased VEGF levels by the amnion (57 ± 26 pg/g protein), as compared with the control group (153 ± 62 pg/g protein) (p < 0.05). On the other hand, MgSO4 significantly increased VEGF levels by the placental villous and the decidua (58 ± 15 pg/g protein, 70 ± 29 pg/g protein; respectively), as compared with the control group (29 ± 11 pg/g protein, 33 ± 14 pg/g protein; respectively) (p < 0.01, p < 0.05; respectively). Exposure to MgSO4 did not affect VEGF levels in normotensive placentas. Conclusion: Reduced levels of VEGF are expressed by some placental compartments in preeclampsia compared with normotensive pregnancy. Perfusion with MgSO4 affects VEGF expression differently by preeclamptic and normotensive placentas. Increased production of placental VEGF in preeclampsia may play a role in the therapeutic action of MgSO4.

Lipopolysaccharide differently affects prostaglandin E2 levels in fetal and maternal compartments of perfused human term placenta

The aim of the present study was to examine the effect of lipopolysaccharide (LPS) on the levels of prostaglandin E(2) (PGE(2)) in the perfusates of the fetal and the maternal compartments of perfused human term placental tissue. Term placentas were perfused for 10h in the absence [control, (n=4)] and presence of LPS [LPS=1 microg/kg perfused placental tissue, (n=4)] in the maternal reservoir. Perfusate samples from the fetal and the maternal circulations were collected every 30 min and examined for PGE(2) levels by radio-immunoassay. PGE(2) levels in the fetal circulation were gradually increased reaching significant peak value of 479+/-159 pg/ml, as compared to PGE(2) levels in the maternal circulation (140+/-146 pg/ml) (p<0.05). After 10 hours of perfusion with control medium, PGE(2) levels in the maternal circulation (347+/-144 pg/ml) were significantly higher as compared to the fetal circulation (150+/-57 pg/ml) (p<0.05). In presence of LPS, PGE(2) levels in the fetal circulation increased reaching a peak value of 1028+/-663 pg/ml after 240 min of perfusion. The levels of PGE(2) in the control group after 240 min of perfusion were significantly lower (156+/-77 pg/ml) (p<0.05). No significant differences were detected in the levels of PGE(2) in the perfusate of the maternal compartment in presence of LPS, as compared to control. Our results suggest that the placenta may play an important role in maintaining high levels of PGE(2) in the fetal circulation and low PGE(2) levels in the maternal circulation during normal pregnancy. Moreover, placental PGE(2) release into the fetal and the maternal circulations may be differently affected in presence of intra-uterine infection/inflammation.

Magnesium Role in Cytokine Regulation of Hypoxic Placentas Related to Certain Placental Pathology

This chapter deals with the issue of magnesium and placenta. Our group studied the area of proinflammatory cytokines in phathological conditions during pregnancy and the influence of MgSO4.