Oxidative stress-induced S100B protein from placenta and amnion affects soluble Endoglin release from endothelial cells.
Oxidative stress with elevated intracellular Ca(2+) concentration as well as endothelial dysfunction is a component of pre-eclampsia. Our aim was to investigate the oxidative stress-dependent expression of Endoglin and Ca(2+)-binding S100B protein from villous and amniotic tissue cultures, and to assess sEng expression from S100B protein-stimulated endothelial cells. We initially examined Endoglin and Hydroxy-nonenal-(HNE)-modified proteins in the placentas and amnion obtained from women with pre-eclampsia (n = 8), and healthy controls (n = 8) by immunohistochemistry. To examine oxidative stress and the S100B protein effect on sEng expression from endothelial cells, normal villous and amniotic tissue cultures were stimulated by 4-HNE, sodium fluoride and xanthine/xanthine oxidase, whereas human umbilical vein endothelial cell cultures were treated with S100B protein in a dose- and time-dependent manner at 37 degrees C in an environment of 95% air and 5% of CO(2). Culture supernatants were assessed using ELISA. Cell viability was determined using MTS assay. The concentrations of sEng and S100B protein were significantly increased in the villous and amniotic tissue culture supernatants under oxidative stress. S100B protein-stimulated endothelial cells released sEng into conditioned media with a significantly higher expression levels at a concentration of 200 pM-20 nM S100B by 2 h, whereas treated with 200 nM of S100B endothelial cells significantly expressed sEng by 12 h and stimulated the cell proliferation by the same period of time. Our findings show that oxidative stress affects sEng and S100B protein expression from villous and amniotic tissues, and picomolar and low nanomolar concentrations of S100B protein significantly up-regulate sEng release from endothelial cells leading to endothelial dysfunction.
Tskitishvili
E
E
Department of Obstetrics and Gynecology, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita City, Osaka 565-0871, Japan. ekaterinet@hotmail.com
Sharentuya
N
N
Temma-Asano
K
K
Mimura
K
K
Kinugasa-Taniguchi
Y
Y
Kanagawa
T
T
Fukuda
H
H
Kimura
T
T
Tomimatsu
T
T
Shimoya
K
K
D000328:Adult; D000447:Aldehydes / Q000494:pharmacology; D000650:Amnion / Q000378:metabolism*; D015703:Antigens, CD / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015237:Dinoprost / Q000031:analogs & derivatives / Q000378:metabolism; D000071063:Endoglin; D042783:Endothelial Cells / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D009414:Nerve Growth Factors / Q000378:metabolism / Q000494:pharmacology* / Q000502:physiology*; D018384:Oxidative Stress / Q000235:genetics / Q000502:physiology*; D010920:Placenta / Q000378:metabolism*; D011225:Pre-Eclampsia / Q000378:metabolism; D011247:Pregnancy; D011956:Receptors, Cell Surface / Q000378:metabolism*; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins / Q000378:metabolism / Q000494:pharmacology* / Q000502:physiology*; D046509:Tissue Culture Techniques; D019820:Xanthine / Q000494:pharmacology; D014969:Xanthine Oxidase / Q000494:pharmacology; D055815:Young Adult
D000447:Aldehydes; D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D009414:Nerve Growth Factors; D011956:Receptors, Cell Surface; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; C579160:S100B protein, human; C462359:tert-4-hydroxy-2-nonenal; D019820:Xanthine; C075750:8-epi-prostaglandin F2alpha; D015237:Dinoprost; D014969:Xanthine Oxidase