PMID- 23731686
OWN - NLM
STAT- MEDLINE
DCOM- 20131107
LR  - 20211021
IS  - 1095-8673 (Electronic)
IS  - 0022-4804 (Print)
IS  - 0022-4804 (Linking)
VI  - 184
IP  - 1
DP  - 2013 Sep
TI  - Regulation of tumor necrosis factor-alpha-induced microvascular endothelial cell 
      hyperpermeability by recombinant B-cell lymphoma-extra large.
PG  - 628-37
LID - S0022-4804(13)00448-4 [pii]
LID - 10.1016/j.jss.2013.04.079 [doi]
AB  - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha), a cytotoxic cytokine, induces 
      endothelial cell barrier dysfunction and microvascular hyperpermeability, leading 
      to tissue edema, a hallmark of traumatic injuries. The objective of the present 
      study was to determine whether B-cell lymphoma-extra large (Bcl-xL), an 
      antiapoptotic protein, would regulate and protect against TNF-alpha-mediated 
      endothelial cell barrier dysfunction and microvascular hyperpermeability. 
      METHODS: Rat lung microvascular endothelial cells were grown as monolayers on 
      Transwell membranes, and fluorescein isothiocyanate-bovine albumin flux (5 mg/mL) 
      across the monolayer was measured fluorometrically to indicate changes in 
      monolayer permeability. The rat lung microvascular endothelial cell adherens 
      junctional integrity and actin cytoskeleton was studied using beta-catenin 
      immunofluorescence and rhodamine phalloidin dye, respectively. Pretreatment of 
      caspase-8 inhibitor (Z-IETD-FMK, 100 muM) for 1 hour and transfection of 
      Bcl-2-homology domain 3-interacting domain death agonist small interfering RNA 
      (10 muM) for 48 hours were performed to study their respective effects on 
      TNF-alpha-induced (10 ng/mL; 1-hour treatment) monolayer permeability. Recombinant 
      Bcl-xL protein (2.5 mug/ml) was transfected in rat lung microvascular endothelial 
      cells for 1 hour, and its effect on permeability was demonstrated using a 
      permeability assay. Caspase-3 activity was assayed fluorometrically. RESULTS: 
      Z-IETD-FMK pretreatment protected the adherens junctions and decreased 
      TNF-alpha-induced monolayer hyperpermeability. Bcl-2-homology domain 3-interacting 
      domain death agonist small interfering RNA transfection attenuated the 
      TNF-alpha-induced increase in monolayer permeability. Recombinant Bcl-xL protein 
      showed protection against TNF-alpha-induced actin stress fiber formation, an increase 
      in caspase-3 activity, and monolayer hyperpermeability. CONCLUSIONS: Our results 
      have demonstrated the protective effects of recombinant Bcl-xL protein against 
      TNF-alpha-induced endothelial cell adherens junction damage and microvascular 
      endothelial cell hyperpermeability. These findings support the potential for 
      Bcl-xL-based drug development against microvascular hyperpermeability and tissue 
      edema.
CI  - Copyright (c) 2013 Elsevier Inc. All rights reserved.
FAU - Sawant, Devendra A
AU  - Sawant DA
AD  - Department of Surgery, Morehouse School of Medicine, Atlanta, Georgia 30310, USA.
FAU - Tharakan, Binu
AU  - Tharakan B
FAU - Wilson, Rickesha L
AU  - Wilson RL
FAU - Stagg, Hayden W
AU  - Stagg HW
FAU - Hunter, Felicia A
AU  - Hunter FA
FAU - Childs, Ed W
AU  - Childs EW
LA  - eng
GR  - K01 HL076815/HL/NHLBI NIH HHS/United States
GR  - T35 HL007815/HL/NHLBI NIH HHS/United States
GR  - 1K01HL07815-01A1/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20130523
PL  - United States
TA  - J Surg Res
JT  - The Journal of surgical research
JID - 0376340
RN  - 0 (BH3 Interacting Domain Death Agonist Protein)
RN  - 0 (Bcl2l1 protein, rat)
RN  - 0 (Bid protein, rat)
RN  - 0 (Ctnnb1 protein, rat)
RN  - 0 (Cysteine Proteinase Inhibitors)
RN  - 0 (Oligopeptides)
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - 0 (bcl-X Protein)
RN  - 0 (benzyloxycarbonyl-isoleucyl-glutamyl-threonyl-aspartic acid fluoromethyl 
      ketone)
RN  - 0 (beta Catenin)
RN  - EC 3.4.22.- (Casp3 protein, rat)
RN  - EC 3.4.22.- (Casp8 protein, rat)
RN  - EC 3.4.22.- (Caspase 3)
RN  - EC 3.4.22.- (Caspase 8)
SB  - IM
MH  - Adherens Junctions/drug effects/metabolism
MH  - Animals
MH  - BH3 Interacting Domain Death Agonist Protein/genetics/metabolism
MH  - Capillary Permeability/drug effects/physiology
MH  - Caspase 3/metabolism
MH  - Caspase 8/metabolism
MH  - Cells, Cultured
MH  - Cysteine Proteinase Inhibitors/pharmacology
MH  - Edema/*metabolism/pathology
MH  - Endothelial Cells/cytology/drug effects/*metabolism
MH  - Lung/cytology
MH  - Oligopeptides/pharmacology
MH  - RNA, Small Interfering/genetics
MH  - Rats
MH  - Recombinant Proteins/metabolism/pharmacology
MH  - Tumor Necrosis Factor-alpha/*metabolism
MH  - bcl-X Protein/*metabolism/pharmacology
MH  - beta Catenin/metabolism
PMC - PMC3759616
MID - NIHMS477698
OTO - NOTNLM
OT  - Adherens junction
OT  - Apoptotic signaling
OT  - BID
OT  - Bcl-2-homology domain 3-interacting domain death agonist
OT  - Vascular permeability
OT  - beta-Catenin
EDAT- 2013/06/05 06:00
MHDA- 2013/11/08 06:00
PMCR- 2014/09/01
CRDT- 2013/06/05 06:00
PHST- 2012/12/20 00:00 [received]
PHST- 2013/04/18 00:00 [revised]
PHST- 2013/04/30 00:00 [accepted]
PHST- 2013/06/05 06:00 [entrez]
PHST- 2013/06/05 06:00 [pubmed]
PHST- 2013/11/08 06:00 [medline]
PHST- 2014/09/01 00:00 [pmc-release]
AID - S0022-4804(13)00448-4 [pii]
AID - 10.1016/j.jss.2013.04.079 [doi]
PST - ppublish
SO  - J Surg Res. 2013 Sep;184(1):628-37. doi: 10.1016/j.jss.2013.04.079. Epub 2013 May 
      23.