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Shock. 2010 Apr;33(4):392-8. doi: 10.1097/SHK.0b013e3181be3e99.

Targeted mutation of Cav-1 alleviates the effect of endotoxin in the inhibition of ET-1-mediated eNOS activation in the liver.

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  • 1Department of Biology, University of North Carolina at Charlotte, Charlotte, North Carolina 28223, USA.

Abstract

Dysfunction of hepatic microcirculation during inflammatory stress conditions is associated with overexpression of caveolin 1 (Cav-1) in sinusoidal endothelial cells. Because Cav-1 binds and inhibits eNOS, it was suggested that Cav-1 overexpression inhibits endothelin 1 (ET-1)-mediated eNOS activation after endotoxemia in the liver; however, a causal link between stress-mediated suppression of eNOS and Cav-1 overexpression has not been fully established. We hypothesize that genetic knockout of Cav-1 reverses the LPS-suppressed ET-1-mediated eNOS activation. In this report, liver sinusoidal endothelial cells (LSECs) from wild-type (WT) and Cav-1 knockout (KO) mice were isolated, pretreated with 100 ng/mL LPS for 6 h, and treated with 10 nmol ET-1 for 30 min. Data showed that LPS increased Cav-1 protein expression (+88%; P < 0.05) and inhibited ET-1-mediated eNOS activation and NO production in WT LSECs. Genetic deletion of Cav-1 increased basal eNOS activity (0.40 in KO vs. 0.15 fmol/min per well in WT; +262%; P < 0.05) and reversed LPS inhibition of ET-1-stimulated eNOS activity (+25.7%; P < 0.05) by increasing eNOS-Ser1177 (+40.3%; P < 0.05) and decreasing eNOS-threonine-495 (-8.8%; P < 0.05) phosphorylation. The reversal of LPS inhibition resulted in an increase in ET-1-induced eNOS translocation to the plasma membrane and an augmentation of NO production in the perinuclear region and plasma membrane of Cav-1 KO LSECs. These results showed that genetic knockout of Cav-1 increased basal eNOS activity and at least partially restored ET-1-mediated eNOS translocation and NO production in LSECs after LPS treatment. In conclusion, Cav-1 overexpression is a requirement for decreased eNOS activity in LSECs after endotoxemia.

PMID:
19730165
[PubMed - indexed for MEDLINE]
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