Mechanism of 17β-estradiol Induced Nitric Oxide Release from Bovine Aortic EndothelialCells / 医药导报

ABSTRACT

Objective To investigate the time and dose dependent effects of 17 β-estradiol ( E2 ) on eNOS phosphorylation and nitric oxide (NO) production from bovine aortic endothelial cells (BAECs). Methods The BAECs were cultured in 24-well flat plate.The dose dependent rapid induction of NO release by E2 in the BAECs was explored,and the non-genomic mechanism was studied. Each experiment was repeated for 3 times. The NO level was quantified by the electron spin resonance spectroscopy(ESR)method,and the phosphorylation of eNOS were determined by Western blot. Results NO release increased time dependently from BAECs after treatment with E2 at 100 nmol?L-1 at 1,5,10 and 15 min,and the effect peaked at 10 min.There were dose dependent effects of NO production as well as eNOS phosphorylation after treatment with E2 at different concentrations for 10 min,and the effect was the most obvious at the concentration of 100 nmol?L-1 .Upon treatment with equal volume of saline,E2 and actinomycin D (25 μg?mL-1 ) for 10 min,the NO release was(5.38±2.35),(10.59±3.28)and(10.68± 3.31) nmol?mg-1 ,respectively,and the eNOS phosphorylation level was 0.36±0.03,0.98±0.08 and 0.99±0.08,respectively. Compared with 0.9% sodium chloride solution,the NO release and eNOS phosphorylation were significantly increased in E2 treated cells(all P<0.05). Conclusion 17 β-estradiol at 100 nmol?L-1 induced eNOS phosphorylation as well as NO production from bovine vascular endothelial cells through non-genomic mechanism.The effect peaked at 10 minutes.