Interaction between regulatory proteins related to the activation of eNOS and CYP4A/20-HETE / 中国药理学通报

ABSTRACT

Aim To investigate the interaction between regulatory proteins related to the activation of eNOS and CYP4A/20-HETE.Methods The endothelia of pulmonary artery of new born bovine were cultured and divided into four groups controlling group ethanol solvent as vehicle (the volume is the same with 1?10~ -6 mol?L~ -1 20-HETE) was added to culture dish and incubated for 10 min; 20-HETE 5 min treatment group1?10~ -6 mol?L~ -1 20-HETE was added to culture dish and incubated for 5 min; 20-HETE 10 min treatment group1?10~ -6 mol?L~ -1 20-HETE was added to culture dish and incubated for 10 min; VEGF 10 min treatment group 1?10~ -8 mol?L~ -1 VEGF was added to culture dish and incubated for 10 min. CYP4A,eNOS and proteins related to the activation of eNOS were measured by Immunoprecipitating and immunobloting.Results Expressions of eNOS and phospho-eNOS(Ser1177)in PAECs were upregulated after the cells were treated with 20-HETE, and similar phenomena were observed when 20-HETE was replaced by VEGF; 20-HETE increased the binding of Hsp、Akt and eNOS.Conclusions ①20-HETE phosphorylates eNOS at Ser1177 site which activates the eNOS to catalyze L-aginine to produce nitric oxide.②eNOS binds with Hsp90 and Akt in endothelial cell of pulmonary artery and 20-HETE increase the binding. ③eNOS binds with CYP4A in endothelial cell of pulmonary artery.