A cellular model for screening neuronal nitric oxide synthase inhibitors

作者：Fang Jianguo; Silverman Richard B^*

来源：Analytical Biochemistry, 2009, 390(1): 74-78.

DOI：10.1016/j.ab.2009.04.004

• Nitric oxide
• Cell-based assay
• Nitric oxide synthase inhibitor
• HEK 293T cells
• Griess reagent

Nitric oxide synthase (NOS) inhibitors are potential drug candidates because it has been well demonstrated that excessive production of nitric oxide critically contributes to a range of diseases. Most inhibitors have been screened in vitro using recombinant enzymes, leading to the discovery of a variety of potent compounds. To make inhibition studies more physiologically relevant and bridge the gap between the in vitro assay and in vivo studies, we report here a cellular model for screening NOS inhibitors. Stable transformants were generated by overexpressing rat neuronal NOS in HEK 293T cells. The enzyme was activated by introducing Calcium ions into cells, and its activity was assayed by determining the amount of nitrite that was formed in Culture medium using the Griess reagent. We tested a few NOS inhibitors with this assay and found that the method is sensitive, versatile, and easy to use. The cell-based assay provides more information than in vitro assays regarding the bioavailability of NOS inhibitors, and it is Suitable for high-throughput screening.

• 出版日期2009-7-1
• 单位西北大学