Functional proteomics of neurokinin B in the placenta indicates a novel role in regulating cytotrophoblast antioxidant defences

Abstract

Neurokinin B (NKB) has recently been demonstrated to be secreted from the placenta in abnormally high amounts in preeclampsia (PE) and to cause hypertension in rats, suggesting it may be a mediator of some pathophysiological features of PE. It is also known that NKB receptors exist in the placenta. To determine the effect of high levels of NKB on the placenta, we have performed proteomics on five separate preparations of cultured purified human term cytotrophoblast cells. The results showed a statistically significant decrease in 20 proteins, of which five were unknown proteins. Proteins important in antioxidant defenses that decreased were thioredoxin, cyclophilin A, cytokeratin 1, and peroxiredoxin 5. Two proteins that inhibit intravascular anticoagulation, cytokeratin 1 and annexin 11 were also decreased. Pathways involving pro-inflammatory cytokine activation of NF-kappa B are opposed by Raf kinase inhibitor protein, which was also decreased. Cofilin 1, a protein involved in defense against bacteria, was also decreased. Among other proteins that were suppressed by NKB were proteasome proteins, desmoplakin, and calgizzarin. Western blots confirmed the decrease in cytokeratin 1 and cyclophilin A protein after NKB exposure. In PE, there is reduced antioxidant activity and increased intravascular coagulation. The findings that high levels of NKB, similar to those observed in PE, can impair these two classes of activity support the hypothesis that high NKB levels may contribute to the pathogenesis of PE.

Overexpressed Cyclophilin A in Cancer Cells Renders Resistance to Hypoxia- and Cisplatin-Induced Cell Death

“…3C and D). Accordingly, evidence that CypA functions as an antioxidant has been recently accumulated (16,(30)(31)(32). Our previous study also showed that overexpressed CypA removed ROS generated by cyclosporin A (17).…”

Proteome Differences in the First- and Third-Trimester Human Placentas

“…9 Up to now, only a few proteomic studies have been performed on the placental tissue or cell lines and the relationship between the placental proteome changes and pregnancy-related disorders, such as pregnancy loss and preeclampsia. 8,[10][11][12][13][14][15][16][17] Considering the scarcity of the published studies on the proteome changes during fetus development, the present study aimed to compare the total placental proteome differences between the normal first-and third-trimester human placentas. In order to fulfill this goal, placental proteins from normal first-and third-trimester human placentas were extracted and subjected to 2D-PAGE.…”

Growth hormone (GH) action in the developing lung: Changes in lung proteins after adenoviral GH overexpression

“…Lung proteins were separated by twodimensional gel electrophoresis, as detailed by Sawicki et al (2003). Protein (200 g) was applied to 11 cm IPG (linear pH gradient range 3-10) strips (six samples on six separate strips for each treatment group; Bio-Rad), with rehydration, for 16 -18 hr at 20°C.…”