Influence of lipopolysaccharide on the permeability of rat brain microvascular endothelial cells and the molecular mechanism.
- Author:
Xiao-Lu DENG
1
;
Fang HE
;
Jing PENG
;
Li-Fen YANG
;
Ci-Liu ZHANG
;
Qiu-Lian XIANG
;
Li-Wen WU
;
Guo-Li WANG
;
Fei YIN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Brain; blood supply; Capillary Permeability; drug effects; Electric Impedance; Endothelial Cells; drug effects; metabolism; Guanine Nucleotide Exchange Factors; analysis; Lipopolysaccharides; pharmacology; Rats; Rats, Sprague-Dawley; Rho Guanine Nucleotide Exchange Factors; Tight Junctions; chemistry; rhoA GTP-Binding Protein; analysis
- From: Chinese Journal of Contemporary Pediatrics 2011;13(11):908-911
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the influence of lipopolysaccharide (LPS) on the permeability of rat brain microvascular endothelial cells (BMECs) and possible molecular mechanism.
METHODSMonolayers of primary rat BMECs were separated and cultured, and then treated with (LPS group) or without LPS (control group). The barrier integrity was measured by transendothelial electrical resistance (TEER) assay. The degrees of RhoA activation were determined by Pull-down assay. The expression levels of p115RhoGEF, zonula occludens-1 (ZO-1), occludin and claudin-5 proteins were detected by Western blot analysis.
RESULTSThe average TEER values of rat BMECs in the LPS group were 108.3±4.2 Ω•cm2 and 85.4±2.5 Ω•cm2 respectively 3 and 12 hrs after LPS treatment, which were significantly lower than that in the control group (159.0±8.6 Ω•cm2). Compared with the control group, the activity of RhoA started to increase 5 minutes after LPS treatment, and the expression of p115RhoGEF protein started to increase 1 hr after LPS treatment and the cellular protein levels of ZO-1, occludin and claudin-5 decreased significantly 3 hrs after LPS treatment in the LPS group (P<0.05).
CONCLUSIONSLPS may activate the p115RhoGEF/RhoA pathway and decrease protein expression of ZO-1, occludin and claudin-5, resulting in an increased permeability of rat BMECs.
