Chase R Cawyer1, Darijana Horvat1, Dean Leonard2, Steven R Allen1, Richard O Jones1, David C Zawieja3, Thomas J Kuehl1, Mohammad N Uddin4. 1. Department of Obstetrics and Gynecology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX. 2. Prehealth Studies, Baylor University, Waco, TX. 3. Department of Medical Physiology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX. 4. Department of Obstetrics and Gynecology, Scott and White Healthcare/Texas A&M Health Science Center College of Medicine, Temple, TX. Electronic address: mnuddin@sw.org.
Abstract
OBJECTIVE: Diabetes mellitus is a risk factor for preeclampsia. Cytotrophoblast (CTB) invasion is facilitated from the conversion of plasminogen to plasmin by urokinase plasminogen activator (uPA), regulated by plasminogen activator inhibitor 1 (PAI-1), and may be inhibited in preeclampsia. This study assessed signaling mechanisms of hyperglycemia-induced CTB dysfunction. STUDY DESIGN: Human CTBs were treated with 45, 135, 225, 495, or 945 mg/dL glucose for 48 hours. Some cells were pretreated with a p38 inhibitor (SB203580) or a peroxisome proliferator-activated receptor-gamma (PPAR-γ) ligand (rosiglitazone). Expression of uPA, PAI-1, and PPAR-γ levels and p38 mitogen-activated protein kinase phosphorylation were measured by Western blot in cell lysates. Messenger ribonucleic acid of uPA and PAI-1 was measured by quantitative polymerase chain reaction. Levels of interleukin-6, angiogenic (vascular endothelial growth factor [VEGF], placenta growth factor [PlGF]) and antiangiogenic factors (soluble fms-like tyrosine kinase-1 [sFlt-1], soluble endoglin [sEng]) were measured in the media by enzyme-linked immunosorbent assay kits. Statistical comparisons were performed using analysis of variance with a Duncan's post-hoc test. RESULTS: Both uPA and PAI-1 protein and messenger ribonucleic acid were down-regulated (P < .05) in CTBs treated with 135 mg/dL glucose or greater compared with basal (45 mg/dL). The sEng, sFlt-1, and interleukin-6 were up-regulated, whereas the VEGF and PlGF were down-regulated by 135 mg/dL glucose or greater. p38 phosphorylation and PPAR-γ were up-regulated (P < .05) in hyperglycemia-treated CTBs. The SB203580 or rosiglitazone pretreatment showed an attenuation of glucose-induced down-regulation of uPA and PAI-1. CONCLUSION: Hyperglycemia disrupts the invasive profile of CTB by decreasing uPA and PAI-1 expression; down-regulating VEGF and PlGF; and up-regulating sEng, sFlt-1, and interleukin-6. Attenuation of CTB dysfunction by SB203580 or rosiglitazone pretreatment suggests the involvement of stress signaling.
OBJECTIVE:Diabetes mellitus is a risk factor for preeclampsia. Cytotrophoblast (CTB) invasion is facilitated from the conversion of plasminogen to plasmin by urokinase plasminogen activator (uPA), regulated by plasminogen activator inhibitor 1 (PAI-1), and may be inhibited in preeclampsia. This study assessed signaling mechanisms of hyperglycemia-induced CTB dysfunction. STUDY DESIGN:HumanCTBs were treated with 45, 135, 225, 495, or 945 mg/dL glucose for 48 hours. Some cells were pretreated with a p38 inhibitor (SB203580) or a peroxisome proliferator-activated receptor-gamma (PPAR-γ) ligand (rosiglitazone). Expression of uPA, PAI-1, and PPAR-γ levels and p38 mitogen-activated protein kinase phosphorylation were measured by Western blot in cell lysates. Messenger ribonucleic acid of uPA and PAI-1 was measured by quantitative polymerase chain reaction. Levels of interleukin-6, angiogenic (vascular endothelial growth factor [VEGF], placenta growth factor [PlGF]) and antiangiogenic factors (soluble fms-like tyrosine kinase-1 [sFlt-1], soluble endoglin [sEng]) were measured in the media by enzyme-linked immunosorbent assay kits. Statistical comparisons were performed using analysis of variance with a Duncan's post-hoc test. RESULTS: Both uPA and PAI-1 protein and messenger ribonucleic acid were down-regulated (P < .05) in CTBs treated with 135 mg/dL glucose or greater compared with basal (45 mg/dL). The sEng, sFlt-1, and interleukin-6 were up-regulated, whereas the VEGF and PlGF were down-regulated by 135 mg/dL glucose or greater. p38 phosphorylation and PPAR-γ were up-regulated (P < .05) in hyperglycemia-treated CTBs. The SB203580 or rosiglitazone pretreatment showed an attenuation of glucose-induced down-regulation of uPA and PAI-1. CONCLUSION:Hyperglycemia disrupts the invasive profile of CTB by decreasing uPA and PAI-1 expression; down-regulating VEGF and PlGF; and up-regulating sEng, sFlt-1, and interleukin-6. Attenuation of CTB dysfunction by SB203580 or rosiglitazone pretreatment suggests the involvement of stress signaling.
Authors: Christina S Han; Melissa A Herrin; Mary C Pitruzzello; Melissa J Mulla; Erika F Werner; Christian M Pettker; Clare A Flannery; Vikki M Abrahams Journal: Am J Reprod Immunol Date: 2014-11-14 Impact factor: 3.886
Authors: Tammy Nguyen; Saunders Lin; Ahmed F Pantho; Belinda M Kohl-Thomas; Madhava R Beeram; David C Zawieja; Thomas J Kuehl; M Nasir Uddin Journal: Mol Cell Biochem Date: 2015-04-12 Impact factor: 3.396
Authors: Yao Ye; Aurelia Vattai; Xi Zhang; Junyan Zhu; Christian J Thaler; Sven Mahner; Udo Jeschke; Viktoria von Schönfeldt Journal: Int J Mol Sci Date: 2017-07-29 Impact factor: 5.923
Authors: Jackson Nteeba; Kaela M Varberg; Regan L Scott; Mikaela E Simon; Khursheed Iqbal; Michael J Soares Journal: BMJ Open Diabetes Res Care Date: 2020-06
Authors: Juan F Mejia; Kelsey M Hirschi; Kary Y F Tsai; Matthew G Long; Benton C Tullis; Eliza E K Bitter; Benjamin T Bikman; Paul R Reynolds; Juan A Arroyo Journal: Reprod Biol Endocrinol Date: 2019-10-23 Impact factor: 5.211