title: [A study of inhibitory effect of chromogranin A derived peptide CGA(47 _ 66) on hyper - permeability of endothelium induced by serum of septic patient].
abstract: OBJECTIVE
  To explore the role of chromogranin A ( CGA) derived peptide CGA47~ ( Chromfungin, CHR) on septic serum induced high permeability of vascular endothelial cells.
  
  
  METHODS
  Human umbilical venous endothelial cell line (EA.hy926 cells) was exposed to CHR, serum of septic shock patient, and tumor necrosis factor-a (TNF-a) respectively. Methyl thiazolyl tetrazolium (MTT) method, Transwell assay and immunofluorescence were performed to determine cell viability (absorbance (A) value J, permeability of monolayer endothelial cells (A value) , and the morphological characteristic and distribution ofF -actin respectively.
  
  
  RESULTS
  Compared with the blank control group, when EA.hy926 were exposed to CHR with 1, 10, 100 nmol/L the cell activity was not significantly affected (A value: 1.219 ± 0.253, 1.179 ± 0.065, 1.179 ± 0.062 vs. 1.306 ± 0.162, all P>0.05), while when the cells was exposed to CHR in 1 000 nmol/L the cell activity was significantly inhibited (A value: 1.049 ± 0.256 vs. 1.306 ± 0.162, t=-2.390, P=0.031 ). Compared with blank control group, when the cells were exposed to CHR of 1, 10, 100 nmol/L a significant decrease in permeability in EA.hy926 cells was observed (A value: 1.619 ± 0.324, 1.496 ± 0.356, 1.132 ± 0.280 vs. 2.315 ± 0.440, P<0.05 or P<0.01 ). Treatment of septic shock patient's serum or TNF-a to EA. hy926 produced an obvious increase in its permeability (septic serum group A value: 1.204 ± 0.248 vs. 0.277 ± 0.017, P<0.01; TNF-a group A value: 2.485 ± 0.113 vs. 1.602 ± 0.679, P<0.05). High-permeability induced by TNF-a or septic shock patient's serum was alleviated hy CHR in the concentration of 1, 10, 100 nmol/L in a dose-dependent manner (septic serum + CHR group A value: 0.299 ± 0.065, 0.224 ± 0.028, 0.131 ± 0.015 vs. 1.204 ± 0.248; TNF -a + CHR group A value: 1.995 ± 0.394, 1.920 ± 0.096, 1.744 ± 0.475 vs. 2.485 ± 0.113, P<0.05 or P<0.01 ). Under a laser scanning confocal microscope, it was found that the F-actin cytoskeleton of EA.hy926 cells was redistributed, and more stress fibers were found in the septic shock patient's serum group and TNF-α group, while CHR obviously alleviated the above effects induced by septic shock patient's serum or TNF-α.
  
  
  CONCLUSIONS
  In a dose-dependent manner, CHR may inhibit increased permeability of vascular endothelial cells induced by septic shock patient's serum, its underlying mechanism may be related to inhibition of the effect of TNF-α.
authors.forename: Ni-na
authors.initials: NN
authors.lastname: Gu
authors.forename: Dan
authors.initials: D
authors.lastname: Zhang
authors.forename: Li
authors.initials: L
authors.lastname: Luo
authors.forename: Xiao-ying
authors.initials: XY
authors.lastname: Chen
authors.forename: Ming
authors.initials: M
authors.lastname: Xie
authors.forename: Jing-lun
authors.initials: JL
authors.lastname: Liu
authors.forename: Li-ping
authors.initials: LP
authors.lastname: Jiang
mesh_terms: D002463:Cell Membrane Permeability / Q000187:drug effects*; D002478:Cells, Cultured; D053379:Chromogranin A / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology; D006801:Humans; D010446:Peptide Fragments / Q000494:pharmacology*; D018805:Sepsis / Q000097:blood; D044967:Serum; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D053379:Chromogranin A; D010446:Peptide Fragments; D014409:Tumor Necrosis Factor-alpha; C437333:chromofungin