cytochrome-c-t and Pre-Eclampsia

Pre-eclampsia (PE) can endanger the survival of the mother and fetus. Currently, the pathogenesis of PE is not completely understood and no fundamental therapeutics are available. The present study was performed to determine the function of miR-128a in HTR-8/SVneo trophoblast cells and to ascertain its underlying role in the pathogenesis of PE.. We investigated the function of miR-128a in HTR-8/SVneo cells by overexpressing. We analyzed the apoptosis of HTR-8/SVneo cells by performing apoptosis assays and measured the loss of mitochondrial membrane potential (Δym), the generation of reactive oxygen species (ROS) and caspase activity. In addition, miR-128a target genes were predicted.. Using computer-based programs, we identified Bax as a direct target of miR-128a. In the apoptosis assays of HTR-8/SVneo cells, miR-128a decreased the Δψm, depleted ATP levels and increased ROS generation, cytochrome c release as well as caspase activation. Further studies showed that miR-128a induced the apoptosis of HTR-8/SVneo cells by down-regulating Bax through the mitochondrial apoptosis pathway.. miR-128a is an up-regulated miRNA in patient with PE. Our study demonstrated that the miR-128a-induced apoptosis of HTR-8/SVneo cells may contribute to PE and miR-128a may be a novel potential therapeutic target for PE.

Topics: Adenosine Triphosphate; Adult; Apoptosis; Base Sequence; bcl-2-Associated X Protein; Caspases; Cytochromes c; DNA, Mitochondrial; Female; Gene Dosage; Gene Expression Regulation; Humans; Membrane Potential, Mitochondrial; MicroRNAs; Mitochondria; Pre-Eclampsia; Pregnancy; Reactive Oxygen Species; Real-Time Polymerase Chain Reaction; Trophoblasts

Antiphospholipid antibodies (aPL) are the strongest maternal risk factor for pre-eclampsia, a hypertensive disease of human pregnancy. Pre-eclampsia is triggered by a toxic factor released from the placenta that activates the maternal endothelium. Antiphospholipid antibodies cause the release of necrotic trophoblast debris from the placental syncytiotrophoblast and this debris can activate endothelial cells. In this study, we investigated how aPL affects syncytiotrophoblast death and production of necrotic trophoblast debris by examining the interaction between aPL and human first trimester placental explants. Human polyclonal and murine monoclonal aPL, but not control antibodies, were rapidly internalised by the syncytiotrophoblast. Inhibitors of endocytosis or the low-density lipoprotein receptor (LDLR) family, but not toll-like receptors, decreased the internalisation of aPL and prevented the release of necrotic trophoblast debris from the syncytiotrophoblast. Once internalised, aPL increased inner mitochondrial membrane leak and Cytochrome c release while depressing oxidative flux through Complex IV of the electron transport system in syncytiotrophoblast mitochondria. These data suggest that the human syncytiotrophoblast internalises aPL by antigen-dependent endocytosis involving LDLR family members. Once internalised by the syncytiotrophoblast, aPL affects the death-regulating mitochondria, causing extrusion of necrotic trophoblast debris which can activate maternal endothelial cells thereby contributing to the pathogenesis of pre-eclampsia.

Topics: Antibodies, Antiphospholipid; Antibodies, Monoclonal; Apoptosis; beta-Cyclodextrins; Cells, Cultured; Chloroquine; Cytochromes c; Electron Transport Complex IV; Endocytosis; Endothelial Cells; Female; Humans; Mitochondrial Membranes; Necrosis; Nitrobenzoates; Organ Culture Techniques; Placenta; Pre-Eclampsia; Pregnancy; Protein Transport; Receptors, LDL; Trophoblasts

Data on respiratory burst activity of granulocytes from healthy and preeclamptic pregnant women are contradictory. To further investigate a possible role of reactive oxygen species in the etiology of preeclampsia, the induced superoxide-anion generation by granulocytes from non-pregnant, healthy pregnant and preeclamptic pregnant women were measured. The reciprocal effects of heat-inactivated and non-inactivated plasma on superoxide production by neutrophils from non-pregnant, healthy pregnant and preeclamptic pregnant subjects were also examined. Superoxide generation was measured by ferricytochrome c reduction. Both phorbol-12.13-dibutirate- and N-formyl-methionyl-leucyl-phenylalanine-induced superoxide production was significantly decreased in normal pregnancy compared to results obtained in non-pregnant and preeclamptic pregnant women. Phorbol-12.13-dibutirate-induced superoxide generation by non-pregnant and preeclamptic neutrophils was significantly inhibited by heat-inactivated and non-inactivated healthy pregnant plasma. N-formyl-methionyl-leucyl-phenylalanine-stimulated superoxide production by non-pregnant and preeclamptic granulocytes was suppressed only by non-inactivated healthy pregnant plasma. Phorbol-12.13-dibutirate-induced superoxide generation of healthy pregnant neutrophils was significantly increased by inactivated and non-inactivated non-pregnant and preeclamptic plasma. N-formyl-methionyl-leucyl-phenylalanine-stimulated superoxide production by healthy pregnant granulocytes was significantly enhanced following treatment of the cells with non-inactivated non-pregnant and preeclamptic pregnant plasma. Deficient superoxide generation in normal pregnancy may be caused by maternal immunosuppressive factors. The failure of reduction in superoxide production in preeclampsia may be partly responsible for endothelial dysfunction. Apart from oxidative stress, a possible role of inefficient maternal immunosuppression should also be considered in the pathogenesis of preeclampsia.

Topics: Adult; Cytochromes c; Female; Gestational Age; Humans; N-Formylmethionine Leucyl-Phenylalanine; Neutrophils; Pre-Eclampsia; Pregnancy; Pregnancy Trimester, Third; Reference Values; Superoxides

Respiratory chain enzymes defect can be found in placentae of some patients with preeclampsia. The purpose of the present study was to evaluate the relationship between respiratory chain enzyme defect and preeclampsia.. Activities of respiratory chain enzymes were measured in 19 patients with preeclampsia (mild 11, and severe 8) and 9 healthy pregnant women as control.. The activities of cytochrome C oxidase (CCO) and adenosine triphosphatase (ATPase) in placentae of severe preeclampsia were significantly lower than those of healthy pregnants (P<0.05). Other enzymes' activities showed no difference among the groups.. Decreasing activities of some respiratory chain enzymes could be one of the pathogenesis of preeclampsia.

Topics: Adenosine Triphosphatases; Alkaline Phosphatase; Cytochromes c; Female; Humans; Placenta; Pre-Eclampsia; Pregnancy

Human placental syncytiotrophoblast is the main barrier for materno-fetal exchange. Analysis of transplacental transport involves the study of ion channels in both the maternal-facing microvillous membrane (MVM) and the fetal-facing basal membrane (BM). Difficulties in having access to intact placenta with conventional electrophysiological methods favour alternative methodologies, such as isolation and reconstitution of membranes in artificial lipid systems. Pre-eclampsia is a major health problem of human pregnancy. The search for altered physiological processes in pre-eclamptic placentae requires the investigation of events at both the microvillous and basal surfaces. The aim of this study was to obtain reliable syncytiotrophoblast plasma membranes from human normal (N) and pre-eclamptic (PE) pregnancies. We describe a protocol which allows for the simultaneous isolation of MVM and BM. The purity of the membranes isolated was evaluated using enzymatic assays, binding studies, Western blotting and immunohistochemistry. Enrichment of alkaline phosphatase activity for MVM was 17 to 21-fold, with 13-16 per cent protein recovery, for both N and PE. Enrichment of adenylate cyclase activity for BM was 9-fold for N, and enrichment of dihydroalprenolol binding to beta-adrenergic receptors was 12-fold for N and 6-fold for PE, with 14 per cent protein recovery for both N and PE. Cross contamination was low and mitochondrial membrane contamination was negligible. We conclude that MVM and BM isolated from placentae of pre-eclamptic women are similar in enrichment and purity to those of healthy women, thus allowing their use in comparative electrophysiological studies.

Topics: Actins; Adenylyl Cyclases; Alkaline Phosphatase; Blotting, Western; Cell Fractionation; Cell Membrane; Centrifugation; Centrifugation, Density Gradient; Cytochromes c; Dihydroalprenolol; Female; Freezing; Humans; Immunohistochemistry; Membrane Proteins; Microscopy, Fluorescence; Mitochondria; Placenta; Pre-Eclampsia; Pregnancy; Protein Binding; Receptors, Adrenergic, beta; Subcellular Fractions; Succinate Dehydrogenase; Trophoblasts

Topics: Cytochromes; Cytochromes a; Cytochromes c; Electron Transport Complex IV; Female; Humans; Oxidoreductases; Placenta; Pre-Eclampsia; Pregnancy