{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T04:37:14Z","timestamp":1787027834234,"version":"build-2736575974"},"reference-count":51,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2024,10,30]],"date-time":"2024-10-30T00:00:00Z","timestamp":1730246400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Scott, Sherwood, and Brindley Foundation, the Department of Obstetrics and Gynecology (MNU)"},{"name":"Noble Centennial Endowment for Research in Obstetrics and Gynecology at Baylor Scott &amp; White Healthcare"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Preeclampsia (preE) is a hypertensive disorder in pregnancies. It is the third leading cause of mortality among pregnant women and fetuses worldwide, and there is much we have yet to learn about its pathophysiology. One complication includes cerebral edema, which causes a breach of the blood\u2013brain barrier (BBB). Urinary marinobufagenin (MBG) is elevated in a preE rat model prior to developing hypertension and proteinuria. We investigated what effect MBG has on the endothelial cell permeability of the BBB. Human brain microvascular endothelial cells (HBMECs) were utilized to examine the permeability caused by MBG. The phosphorylation of ERK1\/2, Jnk, p38, and Src was evaluated after the treatment with MBG. Apoptosis was evaluated by examining caspase 3\/7. MBG \u2265 1 nM inhibited the proliferation of HBMECs by 46\u201350%. MBG induced monolayer permeability, causing a decrease in the phosphorylation of ERK1\/2 and the activated phosphorylation of Jnk, p38, and Src. MBG increased the caspase 3\/7 expression, indicating the activation of apoptosis. Apoptotic signaling or the disruption of endothelia tight junction proteins was not observed when using the p38 inhibitor as a pretreatment in MBG-treated cells. The MBG-induced enhancement of the HBMEC monolayer permeability occurs by the downregulation of ERK1\/2, the activation of Jnk, p38, Src, and apoptosis, resulting in the cleavage of tight junction proteins, and are attenuated by p38 inhibition.<\/jats:p>","DOI":"10.3390\/cells13211800","type":"journal-article","created":{"date-parts":[[2024,10,31]],"date-time":"2024-10-31T06:55:37Z","timestamp":1730357737000},"page":"1800","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Mechanism of Marinobufagenin-Induced Hyperpermeability of Human Brain Microvascular Endothelial Cell Monolayer: A Potential Pathogenesis of Seizure in Preeclampsia"],"prefix":"10.3390","volume":"13","author":[{"given":"Ahmed F.","family":"Pantho","sequence":"first","affiliation":[{"name":"Artemis Biotechnologies LLC, Temple, TX 76502, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manisha","family":"Singh","sequence":"additional","affiliation":[{"name":"Neonatal and Perinatal Medicine, Baylor Scott & White Hospital, Temple, TX 75182, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Syeda H.","family":"Afroze","sequence":"additional","affiliation":[{"name":"Artemis Biotechnologies LLC, Temple, TX 76502, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelsey R.","family":"Kelso","sequence":"additional","affiliation":[{"name":"Obstetrics & Gynecology, Baylor Scott & White Hospital, Temple, TX 75182, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica C.","family":"Ehrig","sequence":"additional","affiliation":[{"name":"Obstetrics & Gynecology, Baylor Scott & White Hospital, Temple, TX 75182, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7077-9895","authenticated-orcid":false,"given":"Niraj","family":"Vora","sequence":"additional","affiliation":[{"name":"Neonatal and Perinatal Medicine, Baylor Scott & White Hospital, Temple, TX 75182, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas J.","family":"Kuehl","sequence":"additional","affiliation":[{"name":"Artemis Biotechnologies LLC, Temple, TX 76502, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven R.","family":"Lindheim","sequence":"additional","affiliation":[{"name":"Neonatal and Perinatal Medicine, Baylor Scott & White Hospital, Temple, TX 75182, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1170-7805","authenticated-orcid":false,"given":"Mohammad N.","family":"Uddin","sequence":"additional","affiliation":[{"name":"Artemis Biotechnologies LLC, Temple, TX 76502, USA"},{"name":"Neonatal and Perinatal Medicine, Baylor Scott & White Hospital, Temple, TX 75182, USA"},{"name":"Obstetrics & Gynecology, Baylor Scott & White Hospital, Temple, TX 75182, USA"},{"name":"Texas A&M University College of Medicine, College Station, TX 77807, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"S1237","DOI":"10.1016\/j.ajog.2020.09.037","article-title":"Eclampsia in the 21st century","volume":"226","author":"Sibai","year":"2022","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"674","DOI":"10.1056\/NEJM199609123351102","article-title":"Pregnancy and the risk of stroke","volume":"335","author":"Kittner","year":"1996","journal-title":"N. Engl. J. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"930","DOI":"10.1161\/01.STR.26.6.930","article-title":"For the Stroke in Pregnancy Study Group, Incidence and causes of strokes associated with pregnancy and puerperium","volume":"26","author":"Sharshar","year":"1995","journal-title":"Stroke"},{"key":"ref_4","first-page":"1344","article-title":"MR Angiography of cerebral vasospasm in preeclampsia","volume":"16","author":"Ito","year":"1995","journal-title":"Am. J. Neuroradiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1016\/0022-1759(83)90303-4","article-title":"Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays","volume":"65","author":"Mosmann","year":"1983","journal-title":"J. Immunol. Methods"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1200","DOI":"10.1016\/0002-9378(89)90665-0","article-title":"Preeclampsia: An endothelial cell disorder","volume":"161","author":"Roberts","year":"1989","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"440","DOI":"10.1016\/0002-9378(72)90493-0","article-title":"Plasma and red cell volumes during pregnancy","volume":"112","author":"Chesley","year":"1972","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_8","first-page":"593","article-title":"Plasma volume contraction: A significant factor in both pregnancy-associated hypertension (pre-eclampsia) and chronic hypertension in pregnancy","volume":"48","author":"Gallery","year":"1979","journal-title":"Q. J. Med."},{"key":"ref_9","first-page":"958","article-title":"Plasma volume determination in normal and preeclamptic pregnancies","volume":"151","author":"Hays","year":"1985","journal-title":"Obstet. Gynecol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"C775","DOI":"10.1152\/ajpcell.00084.2009","article-title":"Inhibition of basal p38 or JNK activity enhances epithelial barrier function through differential modulation of claudin expression","volume":"297","author":"Carrozzino","year":"2009","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1064","DOI":"10.1096\/fj.01-0895com","article-title":"Critical involvement of p38 MAP kinase in pertussis toxin-induced cytoskeletal reorganization and lung permeability","volume":"16","author":"Garcia","year":"2002","journal-title":"FASEB J."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1159\/000193220","article-title":"Vascular leak in a rat model of preeclampsia","volume":"30","author":"Uddin","year":"2009","journal-title":"Am. J. Nephrol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"R1726","DOI":"10.1152\/ajpregu.90963.2008","article-title":"Marinobufagenin causes endothelial cell monolayer hyperpermeability by altering apoptotic signaling","volume":"296","author":"Uddin","year":"2009","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1447","DOI":"10.1016\/0140-6736(93)90889-O","article-title":"Pre-eclampsia: More than pregnancy-induced hypertension","volume":"341","author":"Roberts","year":"1993","journal-title":"Lancet"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"H3179","DOI":"10.1152\/ajpheart.01337.2006","article-title":"Apoptotic signaling induces hyperpermeability following hemorrhagic shock","volume":"292","author":"Childs","year":"2007","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1829","DOI":"10.1084\/jem.191.11.1829","article-title":"Signaling via beta (2) integrins triggers neutrophil-dependent alteration in endothelial barrier function","volume":"191","author":"Gautam","year":"2000","journal-title":"J. Exp. Med."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1161\/01.ATV.18.2.227","article-title":"Shear stress activates p60src-Ras-MAPK signaling pathways in vascular endothelial cells","volume":"18","author":"Jalali","year":"1998","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1067\/mob.2002.119638","article-title":"Endothelial junctional protein redistribution and increased monolayer permeability in human umbilical vein endothelial cells isolated during preeclampsia","volume":"186","author":"Wang","year":"2002","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"207","DOI":"10.3727\/095535491820873191","article-title":"Use of an aqueous soluble tetrazolium\/formazen assay for cell growth assays in culture","volume":"3","author":"Cory","year":"1991","journal-title":"Cancer Commun."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"984","DOI":"10.1016\/j.placenta.2005.12.004","article-title":"Marinobufagenin impairs first trimester cytotrophoblast differentiation","volume":"27","author":"Lamarca","year":"2006","journal-title":"Placenta"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1016\/j.placenta.2007.12.009","article-title":"Marinobufagenin inhibits proliferation and migration of cytotrophoblast and CHO cells","volume":"29","author":"Uddin","year":"2008","journal-title":"Placenta"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1042\/bj3500717","article-title":"A new phosphospecific cell-based ELISA for p42\/p44 mitogen-activated protein kinase (MAPK), p38 MAPK, protein kinase B and cAMP-response-element-binding protein","volume":"350","author":"Versteeg","year":"2000","journal-title":"Biochem. J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1177\/108705710200700306","article-title":"Homogeneous time-resolved fluorescence quenching assay (LANCE) for caspase-3","volume":"7","author":"Kavinen","year":"2002","journal-title":"J. Biomol. Screen."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1152\/ajpcell.00098.2007","article-title":"Endogenous and exogenous cardiac glycosides: Their roles in hypertension, salt metabolism, and cell growth","volume":"293","author":"Schoner","year":"2007","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1159\/000088461","article-title":"Involvement of marinobufagenin in a rat model of human preeclampsia","volume":"25","author":"Vu","year":"2005","journal-title":"Am. J. Nephrol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/j.semnephrol.2005.03.010","article-title":"Endogenous cardiac glycosides: Hormones using the sodium pump as signal transducer","volume":"25","author":"Schoner","year":"2005","journal-title":"Semin. Nephrol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"17946","DOI":"10.1074\/jbc.M800958200","article-title":"Examination of the cellular mechanisms by which marinobufagenin inhibits cytotrophoblast function","volume":"283","author":"Uddin","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1067\/mob.2002.125241","article-title":"Cerebral perfusion pressure, and not cerebral blood flow, may be the critical determinant of intracranial injury in preeclampsia: A new hypothesis","volume":"187","author":"Belfort","year":"2002","journal-title":"Am. J. Obstet. Gynecol."},{"key":"ref_29","first-page":"827","article-title":"Cerebrovascular reactivity in normal pregnancy and preeclampsia","volume":"98","author":"Belfort","year":"2001","journal-title":"Obstet. Gynecol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1016\/j.bbrc.2007.11.167","article-title":"Regulation of the paracellular Na+ and Cl- conductances by the NaCl-generated osmotic gradient in a manner dependent on the direction of osmotic gradients","volume":"366","author":"Tokuda","year":"2008","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1111\/j.1447-0756.2007.00547.x","article-title":"Posterior leukoencephalopathy syndrome as a cause of reversible blindness during pregnancy","volume":"33","author":"Onderoglu","year":"2007","journal-title":"J. Obstet. Gynaecol. Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1002\/jnr.20521","article-title":"Hyperosmotic mannitol induces Src kinase- dependent phosphorylation of \u00df-catenin in cerebral endothelial cells","volume":"80","author":"Farkas","year":"2005","journal-title":"J. Neurosci. Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1099","DOI":"10.1038\/jcbfm.2009.30","article-title":"Inhibition of Src activity decreases tyrosine phosphorylation of occludin in brain capillaries and attenuates increase in permeability of the blood-brain barrier after transient focal cerebral ischemia","volume":"29","author":"Takenaga","year":"2009","journal-title":"J. Cereb. Blood Flow. Metab."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1515\/BC.2005.082","article-title":"hDLG\/SAP97, a member of the MAGUK protein family, is a novel caspase target during cell-cell detachment in apoptosis","volume":"386","author":"Gregorc","year":"2005","journal-title":"Biol. Chem."},{"key":"ref_35","first-page":"973","article-title":"Beta-catenin cleavage in non-apoptotic cells with reduced cell adhesion activity","volume":"15","author":"Nakamoto","year":"2005","journal-title":"Int. J. Mol. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1126\/science.281.5381.1312","article-title":"Caspases: Enemies within","volume":"281","author":"Thornberry","year":"1998","journal-title":"Science"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Cohen, T.S., Gray Lawrence, G., Khasgiwala, A., and Margulies, S.S. (2010). MAPK activation modulates permeability of isolated rat alveolar epithelial cell monolayers following cyclic stretch. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0010385"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.exer.2011.09.009","article-title":"Cell signaling in regulation of the barrier integrity of the corneal endothelium","volume":"95","author":"Srinivas","year":"2011","journal-title":"Exp. Eye Res."},{"key":"ref_39","first-page":"1101","article-title":"Amyloid-beta-induced occludin down-regulation and increased permeability in human brain endothelial cells is mediated by MAPK activation","volume":"14","author":"Tai","year":"2010","journal-title":"J. Cell Mol. Med."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"6877","DOI":"10.4049\/jimmunol.166.11.6877","article-title":"The p38 mitogen-activated protein kinase mediates cytoskeletal remodeling in pulmonary microvascular endothelial cells upon intracellular adhesion molecule-1 ligation","volume":"166","author":"Wang","year":"2001","journal-title":"J. Immunol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"2097","DOI":"10.1242\/jcs.01071","article-title":"The specific fates of tight junction proteins in apoptotic epithelial cells","volume":"117","author":"Bojarski","year":"2004","journal-title":"J. Cell Sci."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1093\/ajh\/hpaa142","article-title":"Preeclampsia and Increased Permeability Over the Blood\u2013Brain Barrier: A Role of Vascular Endothelial Growth Receptor 2","volume":"34","author":"Bergman","year":"2021","journal-title":"Am. J. Hypertens."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2412","DOI":"10.2741\/2243","article-title":"Endothelial barrier function in preeclampsia","volume":"12","author":"Wang","year":"2007","journal-title":"Front Biosci."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"R918","DOI":"10.1152\/ajpregu.00499.2013","article-title":"Marinobufagenin regulates permeability and gene expression of brain endothelial cells","volume":"306","author":"Ing","year":"2014","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1055\/s-0031-1272965","article-title":"Marinobufagenin Levels in Preeclamptic Patients: A Preliminary Report","volume":"28","author":"Agunanne","year":"2011","journal-title":"Am. J. Perinatol."},{"key":"ref_46","first-page":"19","article-title":"Endogenous Na\/K-ATPase inhibitors in patients with preeclampsia","volume":"52","author":"Averina","year":"2006","journal-title":"Cell. Mol. Biol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1070","DOI":"10.1210\/jcem-59-6-1070","article-title":"An endogenous ouabain-like factor associated with hypertensive pregnant women","volume":"59","author":"Graves","year":"1984","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1056\/NEJM198802253180815","article-title":"Antidigoxin antibodies in eclampsia","volume":"318","author":"Goodlin","year":"1988","journal-title":"N. Engl. J. Med."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1159\/000169054","article-title":"Elevated endoxin-like factor complicating a multifetal second trimester pregnancy: Treatment with digoxin-binding immunoglobulin","volume":"16","author":"Adair","year":"1996","journal-title":"Am. J. Nephrol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1179","DOI":"10.1097\/00004872-199917080-00018","article-title":"Circulating bufadienolide and cardenolide sodium pump inhibitors in preeclampsia","volume":"17","author":"Lopatin","year":"1999","journal-title":"J. Hypertens."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1080\/15321819.2010.538107","article-title":"A chemifluorescent immunoassay for the determination of marinobufagenin in body fluids","volume":"32","author":"Lai","year":"2011","journal-title":"J. Immunoass. Immunochem."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/13\/21\/1800\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:24:37Z","timestamp":1760113477000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/13\/21\/1800"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,30]]},"references-count":51,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2024,11]]}},"alternative-id":["cells13211800"],"URL":"https:\/\/doi.org\/10.3390\/cells13211800","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,30]]}}}