一种实时检测剪切力引起培养的大鼠动脉内皮细胞内一氧化氮含量的新方法

建立一个稳定和实时检测在不同剪切力作用下内皮细胞内一氧化氮含量的方法。利用流动小室建立内皮细胞剪切模型 ,在内皮细胞用DAF FM染色后 ,用Zeiss荧光共聚焦显微镜和ICCD摄象头检测细胞内的荧光强度。DAF FM的荧光强度可以反映一氧化氮的胞内含量。剪切力引起内皮细胞合成一氧化氮增加 ,并且这种作用是随着剪切力的增加而增加。剪切力的作用被一氧化氮合酶抑制剂L NAME全部抑制 ,被无Ca2 缓冲液部分抑制。这个方法可以实时反映一氧化氮含量的变化 ,可以用来研究剪切力引起一氧化氮变化的机制以及用来评价内皮细胞对剪切力的反应特性

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.