title: Expression of HMGB1 in septic serum induces vascular endothelial hyperpermeability.
abstract: The aim of the present study was to investigate the effects of high mobility group protein B1 (HMGB1), which is expressed in the serum of patients with sepsis, on vascular endothelial permeability. Sera from patients with sepsis were used to treat endothelial cells (ECs), and the effect on endothelial permeability was evaluated using immunofluorescence. The morphologies of endothelial cytoskeletal actin and vascular endothelial (VE)‑cadherin were assessed using laser scanning confocal microscopy. The protein expression levels of HMGB1, B‑cell lymphoma 2 (BCL‑2) and BCL‑2‑associated X protein (BAX) were detected using western blotting. EC apoptosis was measured using flow cytometry. The results demonstrated that HMGB1 was significantly expressed in the serum 24 h following the onset of sepsis, and the expression levels peaked at 48 h, which were sustained until 96 h post‑onset. Compared with the control group, treatment of the ECs with 20% septic serum in vitro significantly increased endothelial monolayer permeability (P<0.01), markedly induced transcellular filamentous (F)‑actin rearrangement with stress fiber formation, and resulted in the localization of VE‑cadherin fragmentations at the cell borders with increased gaps between ECs. Furthermore, flow cytometry showed that the apoptotic rate of ECs was significantly increased following treatment with septic serum. In addition, the expression levels of BAX were significantly increased, whereas the expression levels of BCL‑2 were significantly decreased. Pretreatment with an HMGBI inhibitor (ethyl pyruvate; 5 µM) 24 h prior to treatment with the septic serum attenuated the effects of septic serum treatment. Together, these findings suggested that treatment of ECs with sera from patients with sepsis may induce the loss of vascular endothelial monolayer integrity, elicit the formation of endothelial F‑actin stress fibers and initiate VE‑cadherin redistribution, which may be attributed to high levels of HMGB1 in the serum. This mechanism also appears to involve changes in the activation of BAX and BCL‑2, resulting in EC apoptosis.
authors.affiliation: Emergency Department, Chongming Branch, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 202150, P.R. China.
authors.forename: Yun-Jiang
authors.initials: YJ
authors.lastname: Zheng
authors.affiliation: Administration Division, Chongming Branch, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 202150, P.R. China.
authors.forename: Wei-Ping
authors.initials: WP
authors.lastname: Xu
authors.affiliation: Department of Oncology, Chongming Branch, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 202150, P.R. China.
authors.forename: Gang
authors.initials: G
authors.lastname: Ding
authors.affiliation: Administration Division, Chongming Branch, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 202150, P.R. China.
authors.forename: Yu-Hua
authors.initials: YH
authors.lastname: Gao
authors.affiliation: Emergency Department, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, P.R. China.
authors.forename: Hai-Rong
authors.initials: HR
authors.lastname: Wang
authors.affiliation: Emergency Department, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, P.R. China.
authors.forename: Shu-Ming
authors.initials: SM
authors.lastname: Pan
mesh_terms: D000199:Actins / Q000378:metabolism; D015703:Antigens, CD / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects; D015820:Cadherins / Q000378:metabolism; D002199:Capillary Permeability / Q000187:drug effects; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology*; D005260:Female; D024243:HMGB1 Protein / Q000097:blood*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008297:Male; D008875:Middle Aged; D011773:Pyruvates / Q000494:pharmacology; D018805:Sepsis / Q000097:blood* / Q000503:physiopathology*; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D000199:Actins; D015703:Antigens, CD; D015820:Cadherins; D024243:HMGB1 Protein; D011773:Pyruvates; D051028:bcl-2-Associated X Protein; D000099152:Cadherin 5; C046522:ethyl pyruvate