PMID- 12622935
OWN - NLM
STAT- MEDLINE
DCOM- 20030331
LR  - 20081121
IS  - 0366-6999 (Print)
IS  - 0366-6999 (Linking)
VI  - 115
IP  - 12
DP  - 2002 Dec
TI  - Shear stress induces interleukin-8 mRNA expression and transcriptional activation 
      in human vascular endothelial cells.
PG  - 1838-42
AB  - OBJECTIVE: To examine interleukin-8 (IL-8) mRNA expression induced by flow shear 
      stress in human umbilical vein endothelial cells (HUVECs) and investigate its 
      transcriptional activation. METHODS: Flow laminar shear stress 4.2 dyne/cm(2) was 
      used for the stimulating experiments. The flow shear stress-induced IL-8 mRNA 
      expression was detected by reverse transcription-polymerase chain reaction 
      (RT-PCR). pEGFP1 was used to construct IL-8 reporter gene pEGFP1-IL8USCS for 
      determining IL-8 gene transcriptional activation through gene transfer and flow 
      cytometric analysis. NF-kappaB nuclear translocation was observed by 
      immunocytofluorescent staining. Western blot was used to examine IkappaB 
      phosphorylation and degradation. RT-PCR, Northern blot, immunocytofluorescent 
      staining and laser confocal microscopy were used to determine Toll-like 
      receptor-4 (TLR-4) expression at mRNA and protein levels in the cells. RESULTS: 
      There was a marked increase in IL-8 mRNA expression in HUVECs after 120 min of 
      exposure to laminar flow shear stress. When exposed to shear stress for 180 min, 
      there was an increase in enhanced green fluorescent protein expression in 
      pEGFP1-IL8USCS-transfected endothelial cells. NF-kappaB p65 immunocytofluorescent 
      staining of HUVECs showed that when exposed to the same flow shear stress for 30 
      or 60 min, the cell nuclei became stained; after 90 or 120 min, the staining 
      became much more pronounced. A significant increase in P-IkappaB in the cell 
      lysates occurred after 10 min of exposure while blot density dramatically dropped 
      after 60 min of exposure. The density of the IkappaB blot dropped with increasing 
      exposure time after 30 min. TLR-4 was present on the surface of HUVECs. HUVECs 
      constitutively expressed TLR-2 and TLR-4 mRNA; when exposed to flow shear stress 
      for 60 min, TLR-4 mRNA expression increased. CONCLUSION: NF-kappaB activation is 
      involved in flow shear stress-induced IL-8 mRNA expression in human umbilical 
      vein endothelial cells. TLR-4 receptor for innate immunity most likely mediate 
      these events.
FAU - Liang, Feng
AU  - Liang F
AD  - Research Units of Infection and Immunity, School of Basic Medical Sciences, West 
      China Medical Center, Sichuan University, Chengdu 610041, China.
FAU - Huang, Ning
AU  - Huang N
FAU - Wang, Boyao
AU  - Wang B
FAU - Chen, Huaiqing
AU  - Chen H
FAU - Wu, Lizhi
AU  - Wu L
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Chin Med J (Engl)
JT  - Chinese medical journal
JID - 7513795
RN  - 0 (Drosophila Proteins)
RN  - 0 (Interleukin-8)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (NF-kappa B)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Cell Surface)
RN  - 0 (TLR2 protein, human)
RN  - 0 (TLR4 protein, human)
RN  - 0 (Toll-Like Receptor 2)
RN  - 0 (Toll-Like Receptor 4)
RN  - 0 (Toll-Like Receptors)
SB  - IM
MH  - Blotting, Northern
MH  - Cells, Cultured
MH  - *Drosophila Proteins
MH  - Endothelium, Vascular/cytology/*metabolism
MH  - Humans
MH  - Interleukin-8/*genetics
MH  - Membrane Glycoproteins/analysis
MH  - NF-kappa B/metabolism
MH  - RNA, Messenger/*analysis
MH  - Receptors, Cell Surface/analysis
MH  - Stress, Mechanical
MH  - Toll-Like Receptor 2
MH  - Toll-Like Receptor 4
MH  - Toll-Like Receptors
MH  - *Transcriptional Activation
EDAT- 2003/03/08 04:00
MHDA- 2003/04/01 05:00
CRDT- 2003/03/08 04:00
PHST- 2003/03/08 04:00 [pubmed]
PHST- 2003/04/01 05:00 [medline]
PHST- 2003/03/08 04:00 [entrez]
PST - ppublish
SO  - Chin Med J (Engl). 2002 Dec;115(12):1838-42.