Alterations In Preeclampsia Research Articles

Preeclampsia-associated reduction of cathepsin D activity in the umbilical cord

Background Preeclampsia is accompanied by an increase of collagen contents in the umbilical cord (UC) arteries and in Wharton's jelly. Cathepsin D is one of the enzymes which participates in collagen degradation and activates precursor forms of collagenolytic metalloproteinases. It was decided to evaluate the activity of cathepsin D within umbilical cord arteries, veins and Wharton's jelly and its alterations in preeclampsia. Materials and methods Umbilical cord components were separated and submitted to homogenisation/extraction with 0.05 M Tris–HCl+0.2% Triton X-100, pH 7.5. Proteolytic activities of the extracts were studied with a use of cathepsin D-specific substrate. Western immunoblot technique was employed to detect this enzyme. Results It was found that human umbilical cord tissues contain both active and inactive forms of cathepsin D. Preeclampsia is associated with a distinct increase in the amount of this enzyme in the umbilical cord, whereas its activity deeply decreased. Activation with trypsin augments cathepsin D activity in preeclamptic umbilical cord to the values observed in control arteries or even exceeds the control values (veins, Wharton's jelly). Conclusions Preeclampsia is associated with a reduction in the activity of cathepsin D in human umbilical cord. The low activity of cathepsin D may reduce collagen degradation and enhance its accumulation in the umbilical cord, especially in the arteries. Similar changes in other foetal blood vessels may result in an increase of vascular resistance and hypertension, which may persist after birth.

Collagen of umbilical cord vein and its alterations in pre-eclampsia.

The state of the vascular system of the mother and of placenta is known to exert a great influence on intrauterinal development of the fetus. Pre-eclampsia is the most common pathological syndrome connected with pregnancy. Since collagen is one of the main constituents of the vessel wall a comparison was made with collagen content and its molecular polymorphism in umbilical cord veins of newborns from healthy and pre-eclamptic mothers. It was found that umbilical cord veins of newborns from mothers with pre-eclampsia contained 18% less collagen than those of the newborns from normal pregnancies. This decrease was accompanied by a slight decrease of collagen solubility, but all its types (I, II, IV, V and VI) were present. However, the umbilical vein wall of newborns from mothers with pre-eclampsia contained relatively less of type I and more of type III collagen than the normal umbilical cord. These differences may be connected with a disturbance of blood flow in fetus of a woman with pre-eclampsia.

TGF-β3 Expression During Umbilical Cord Development and its Alteration in Pre-eclampsia

Members of the TGF-β family have been shown to play an important role in numerous tissues during development. In the present study we have investigated the spatial and temporal expression of TGF-β3, in human umbilical cord development. Total TGF-β3 protein content, assessed by immunoblotting, increased with advancing gestation as did immunostaining and mRNA in Wharton’s jelly fibroblasts. Immunohistochemical analysis revealed that TGF-β3 was present in all cell types. Temporal changes in TGF-β3 expression were observed in the vascular smooth muscle cells, such that with advancing gestation TGF-β3 protein expression and became mostly restricted to the extracellular compartment of the vascular media. This was associated with a decrease in TGF-β3 mRNA expression in umbilical vascular smooth muscle cells. Of clinical significance, umbilical cords from pregnancies complicated by pre-eclampsia, showed a significant reduction in total TGF-β3 protein expression when compared to those of age-matched patients. Both TGF-β3 mRNA and protein expression were downregulated in the endothelium and smooth muscle layers of the umbilical arteries, as well as in the Wharton jelly fibroblasts. Our data demonstrate that during umbilical cord development TGF-β3 expression is spatially and temporally regulated and that TGF-β3 expression is altered in umbilical cords of pregnancies complicated by pre-eclampsia. We speculate that the downregulation of TGF-β3 expression found in pre-eclamptic umbilical cord may contribute to the abnormal structure and mechanical properties seen in these pathological umbilical cords.

The use of platelet density and volume measurements to estimate the severity of pre-eclampsia.

This study evaluated whether it is possible to estimate the severity of pre-eclampsia through in vitro measurements of platelet and granulocyte parameters. Eighteen pre-eclamptic women in the third-trimester of pregnancy and 11 women in the third-trimester of normal pregnancies were included in the study. Three to 12 months after delivery, 15 patients with pre-eclampsia and all the subjects with normal pregnancies were re-examined. Before delivery, peak platelet density was determined using a specially designed apparatus. Before and 3-12 months after delivery the following were measured: platelet counts, mean platelet volume and neutrophil and monocyte counts. Furthermore, circulating P-selectin, interleukin-6 and myeloperoxidase were determined to estimate platelet, monocyte and granulocyte activities, respectively. Compared to their results after delivery, pre-eclamptic females demonstrated lower platelet counts (P < 0.001) and raised mean platelet volumes (P < 0.01). Both pre-eclamptic women (P < 0.01) and normal pregnancies (P < 0.05) demonstrated elevated soluble P-selectin at pregnancy. Then pre-eclamptic women had advanced neutrophil counts (P < 0.01) but normal pregnancies showed a similar phenomenon (P < 0.001). Interleukin-6 remained normal during pregnancy. Plasma myeloperoxidase levels were lower both in pre-eclampsia (P < 0.05) and in normal pregnancies (P < 0.001). In pre-eclampsia elevated blood pressure was related to higher mean platelet volumes (P < 0.05). Furthermore, a group of pre-eclamptic females whose platelets had disturbed density distribution displayed elevated mean platelet volumes (P < 0.01). The present work demonstrates considerable platelet alterations in pre-eclampsia. We failed to show granulocyte involvement in the pathogenesis of the disease. Severe pre-eclampsia is related to elevated mean platelet volumes. The latter parameter is associated with disturbed density distribution. It appears possible to estimate disease severity from measurements of platelet density and volume.

Extracellular matrix components of the wall of umbilical cord vein and their alterations in pre-eclampsia

Pre-eclampsia--edema, proteinuria, hypertension (EPH-gestosis) is one of the more common complications observed during pregnancy. The umbilical cord vein walls were taken from newborns delivered by healthy mothers (control material) and by mothers with polysymptomatic pre-eclampsia (investigated material). Normal saphenous vein walls were collected from adult subjects undergoing varicose vein surgery. The collagen content was measured by the assay of hydroxyproline. Elastin was determined according to Fastin Elastin Assay and gravimetrically. Glycosaminoglycans content was determined by uronic acids assay. The collagen content decreased in the pre-eclampsia material. The amount of soluble elastin increased in the investigated material. The insoluble elastin content decreased in the umbilical cord veins of newborns delivered by mothers with pre-eclampsia. Reconstructing the umbilical cord vein wall may disturb fetal blood flow and affect the vascular system in adulthood.

Preeclampsia: The Endothelium, Circulating Factor(s) and Vascular Endothelial Growth Factor

It has been proposed that endothelial cell activation is the primary event in the multisystem disorder of preeclampsia. Evidence for endothelial involvement in this condition abounds. The best-characterized morphologic abnormality of this syndrome, glomerular endotheliosis, involves endothelial cells. Also associated with preeclampsia is a loss of endothelial cell integrity, with the consequent increase in vascular permeability, and an increase in the circulating levels of the endothelial cell markers, fibronectin, von Willebrand factor, tissue plasminogen activator, and plasminogen activator inhibitor-1. It is now well documented that endothelial activation contributes to the coagulation abnormalities observed in this disease. There is much evidence that the endothelial alterations in preeclampsia result from one or more circulating factors. The incubation of cultured endothelial cells with serum or plasma samples, taken from normal pregnant women and women with preeclampsia, results in marked alterations in cell behavior and metabolic processes. More recently, experiments employing myographic techniques have demonstrated convincingly the effects of a circulating factor(s) on the function of endothelial cells of resistance arteries. Vascular endothelial growth factor (VEGF) possesses many of the characteristics required of a candidate circulating factor. It contains a hydrophobic secretory signal sequence, exerts in vitro effects specific to vascular endothelial cell, and promotes endothelial expression of procoagulant activity. Circulating VEGF concentrations are elevated in women with preeclampsia, and VEGF increases microvascular endothelial cell prostacyclin production in a dose-dependent manner, analogous to the acute effects of plasma from patients with preeclampsia. Similarly, in myographic studies, when myometrial resistance arteries are incubated with VEGF, there are dose-dependent alterations in endothelium-dependent behavior, mirroring those found after incubation with plasma from patients with preeclampsia.

Mechanisms of Endothelium-Dependent Relaxation in Myometrial Resistance Vessels and Their Alteration in Preeclampsia

Objective: To investigate the importance of prostacyclin and nitric oxide synthesis in endothelium-dependent relaxation in myometrial resistance vessels, and to test the hypothesis that a deficiency in nitric oxide synthesis contributes to the known alterations in endothelial function in preeclampsia.Methods: Thirty-six women with normal pregnancies and 14 with preeclampsia had the myometrium biopsied at cesarean section. Resistance arteries were dissected and mounted on a wire myograph. After preconstriction with vasopressin, vessels were treated cumulatively with bradykinin. The process was repeated in the presence of indomethacin and then indomethacin and NG-monomethyl-L-arginine (L-NMMA).Results: The vessels from women with normal pregnancies showed endothelium-dependent relaxation to bradykinin which was not significantly altered by the presence of indomethacin. The addition of L-NMMA significantly, but only partially, reduced the relaxation to bradykinin in the presence of indomethacin (p= 0.03).The vessels from women with preeclampsia showed markedly reduced relaxation to bradykinin (p < 0.0001), as compared to vessels from normal pregnant women. Relaxation of vessels from women with preeclampsia was increased by the addition of indomethacin (p = 0.03) but was virtually eradicated by the presence of L-NMMA.Conclusions: Eicosanoid synthesis plays little part in the relaxation of normal pregnant myometrial vessels to bradykinin. Nitric oxide synthesis mediates part but not all of the endothelium-dependent relaxation. In preeclampsia, relaxation to bradykinin is reduced; inhibition of eicosanoid synthesis allows increased relaxation, and nitric oxide synthesis appears to mediate a greater proportion of the relaxation than in normal pregnant women.

Plasma renin activity, angiotensin II, prostacyclin and thromboxane A2 concentrations in 139 preeclamptic patients.

Plasma renin activity, plasma concentrations of angiotensin II (AngII), stable metabolites (6-keto-prostaglandin F1 alpha: 6-keto-PGF1 alpha) of prostacyclin (PGI2) and a metabolite (thromboxane B2: TXB2) of thromboxane A2 (TXA2) were measured with radioimmunoassay(RIA) in 107 normal pregnancy (control) and 139 preeclamptic patients in 28-41 gestational weeks. PRA and 6-keto-PGF1 alpha were significantly higher and AngII was slightly higher in preeclampsia than in control, and TXB2 was significantly lower in preeclampsia in control. The ratio of 6-keto-PGF1 alpha/TXB2 was significantly lower in preeclampsia than in control. These data suggest that the changes in the renin-angiotensin system may not be primary alterations in preeclampsia. It can be speculated that in preeclampsia the changes in absolute concentrations of 6-keto-PGF1 alpha and TXB2 are less important than the decrease in the ratio of the 6-keto-PGF1 alpha/TXB2.

Immunofluorescent studies of the endometrial arteries in the first trimester of pregnancy.

Endometrial spiral arteries from curetted endometrium of 110 first-trimester pregnancies were studied by immunofluorescent (IF) technics using antibodies against human G, M, and A immunoglobulins, C3, C4, and fibrinogen. Heavy deposition of C3 in the arterial walls was found in 16 (14.6%) cases. Immunoglobulins, C4, and fibrinogen were found in only a few cases, and their staining was weak and not considered in this study. There was also a statistically significant (P less than 0.01) higher deposition of C3 in arterial walls of primipara (14 of 52), as compared to multipara (2 of 58). The possible mechanisms of C3 deposition and the importance of the higher incidence of this deposition in primipara are discussed in relation to suggested immunologic pathogenetic alterations in preeclampsia.