Real-time detection of nitric oxide in cultured rat aorta endothelial cells induced by shear stress.
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.
Ye
Jian-Feng
JF
Department of Biomedical Engineering, Purdue University, West Lafayette, IN 47906, USA.
Zheng
Xiao-Xiang
XX
Xu
Lisa Xueming
LX
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
D004791:Enzyme Inhibitors; D009599:Nitroprusside; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D019331:NG-Nitroarginine Methyl Ester