Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (46): 8609-8612.doi: 10.3969/j.issn.1673-8225.2011.46.014

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Effective factors for regulating monolayer permeability of human umbilical vein endothelial cells

Yan Cheng hui, Yu Hai-bo, Zhang Xiao-lin, Kang Jian, Han Ya-ling   

  1. Department of Cardiology, Institute of Cardiovascular Diseases, General Hospital of Shenyang Military Area Command of Chinese PLA, Shenyang  110016, Liaoning Province, China
  • Received:2011-05-08 Revised:2011-08-11 Online:2011-11-12 Published:2011-11-12
  • Contact: Han Ya-ling, Doctor, Professor, Doctoral supervisor, Department of Cardiology, Institute of Cardiovascular Diseases, General Hospital of Shenyang Military Area Command of Chinese PLA, Shenyang 110016, Liaoning Province, China hanyaling@263.net
  • About author:Yan Cheng-hui☆, Doctor, Associate chief technician, Department of Cardiology, Institute of Cardiovascular Diseases, General Hospital of Shenyang Military Area Command of Chinese PLA, Shenyang 110016, Liaoning Province, China yanch1029@163.com
  • Supported by:

    the National Natural Science Foundation of China (General Program), No. 30770793*, 30971218*，81070097*; the National Natural Science Foundation of China (Youth Foundation Program), No. 30800465*; the Natural Science Foundation of Liaoning Province, No. 20092088*; High-Tech Science and Technology Program of Liaoning Province, No. 2009225009-9*

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Abstract:

BACKGROUND: The molecular mechanism underlying exogenous stimulation-caused vascular barrier dysfunction remains poorly understood in the field of vascular pathophysiology.
OBJECTIVE: To investigate the effective factors for regulating monolayer permeability of human umbilical vein endothelial cells and search for the effective treatment target.
METHODS: Human umbilical vein endothelial cells (HUVECs) were exposed to lipopolysaccharide (LPS) with and without specific inhibitors of RhoA and F-actin stress fibre. Changes in RhoA/SRF/F-actin activity in association with cell permeability and F-actin rearrangement signaling were investigated by immunostaining, transwell assay and western blot analysis.
RESULTS AND CONCLUSIONS: When HUVECs were treated with LPS, F-actin cytoskeleton rearrangement was detected by rhodamine-phalloidin staining in a dose and time-dependent manner, meanwhile the HUVEC hyperpermeability was investigated. Furthermore, the RhoA activity and SRF translocalization into nuclei were identified to be associated with elevation of permeability in HUVECs. The effects were blocked when cells were pretreated with Y27632 or Latrunculin B, respectively. Moreover, Latrunculin B added also inhibited the translocalization of SRF into the nuclei mediated by RhoA activity. Our results have identified the RhoA/SRF and their substrates F-actin rearrangement mediated LPS-induced changes in HUVECs permeability.

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