Role of TRPV4 channels in regulation of eNOS expression in brain microvascular endothelial cells under the condition of mechanical stretch.
10.11817/j.issn.1672-7347.2015.09.003
- Author:
Jiujun PU
1
;
Zhiming WANG
1
;
Hui ZHOU
1
;
Ailing ZHONG
1
;
Lunliang RUAN
1
;
Kai JIN
1
;
Gang YANG
1
Author Information
1. Department of Neurosurgery, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China.
- Publication Type:Journal Article
- MeSH:
Brain;
cytology;
Calcium;
metabolism;
Cells, Cultured;
Endothelial Cells;
physiology;
Humans;
Nitric Oxide Synthase Type III;
metabolism;
Stress, Mechanical;
TRPV Cation Channels;
physiology
- From:
Journal of Central South University(Medical Sciences)
2015;40(9):960-966
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To identify the subtype of transient receptor potential (TRPs) channel involved in stretch-induced injury of human brain microvascular endothelial cells (HBMEC) and to explore the mechanism responsible for eNOS expression.
METHODS:TRPs expression was examined by Western blot and immunocytofluoresence in the cultured HBMEC. Mechanical stretch was performed by mini-type multi-functional bio-impact machine. The levels of free calcium ion in cells were examined by the flow cytometry. The eNOS expression was detected by Western blot.
RESULTS:The mRNA and protein expression of TRPV4 was detected in HBMEC by qRT-PCR, Western blot and immunocytofluoresence. The levels of free calcium ion in the stretch-treated HBMEC was significantly decreased in the presence of TRPV4 specific inhibitor (P<0.001), but there was no difference in calcium levels between the stretch and the control or unspecific inhibitor group (P=0.072 or 0.308). The levels of eNOS protein in the stretch-treated HBMEC were reduced in the presence of TRPV4 specific inhibitor or NOS inhibitor (P<0.05), but it was not changed compared with that in the control group (P>0.05).
CONCLUSION:The eNOS expression is up-regulated under the condition of mechanic stretch, which is related to the activation of TRPV4, resulting in the influx of calcium.
