title: Real-time detection of nitric oxide in cultured rat aorta endothelial cells induced by shear stress.
abstract: To establish a stable and real-time method to detect the production of intracellular nitric oxide (NO) of endothelial cells under different shear stresses. After the cultured endothelial cells were loaded with DAF-FM, the relative NO production was determined by fluorescence intensity, which was detected using Zeiss Axioskop 2 fluorescence microscope and ICCD-camera. The fluorescence of DAF-FM can reflect NO production. Shear stress can induce a dose-dependent increase of NO production, and the increase can be totally inhibited by L-NAME and partly inhibited by Ca(2+)-free buffer. The method can be used to detect the change of NO production in real time, and it can also be used to study the mechanism related to NO increase induced by shear stress.
authors.affiliation: Department of Biomedical Engineering, Purdue University, West Lafayette, IN 47906, USA.
authors.forename: Jian-Feng
authors.initials: JF
authors.lastname: Ye
authors.forename: Xiao-Xiang
authors.initials: XX
authors.lastname: Zheng
authors.forename: Lisa Xueming
authors.initials: LX
authors.lastname: Xu
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008856:Microscopy, Fluorescence / Q000379:methods; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000032:analysis*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D009599:Nitroprusside / Q000494:pharmacology; D051381:Rats; D013314:Stress, Mechanical; D013997:Time Factors
chemical_list: D004791:Enzyme Inhibitors; D009599:Nitroprusside; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D019331:NG-Nitroarginine Methyl Ester