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为了建立基于细胞的β_2肾上腺素能受体激动剂筛选体系,将大鼠β_2受体基因克隆人pcDNA3.1载体,转染具有cAMP应答元件调控的绿色荧光蛋白报告基因的中国仓鼠卵巢细胞(CHO),从得到的阳性单克隆细胞中筛选得到一个稳定表达β_2受体,并且在受到激动剂刺激后发出剂量依赖绿色荧光强度的细胞株.将此筛选体系用于从中药中筛选具有β_2受体激动剂活性的化合物.从几种芸香科药材中得到了辛弗林(synephine),从细辛和附子中得到了去甲乌药碱(higenamine),首次从受体水平证明了它们的β_2激动剂活性.此筛选体系适合从大量天然化合物中快速筛选具有β_2激动剂活性的成分.
To establish a cell-based β 2 adrenergic receptor agonist screening system, the rat β 2 receptor gene was cloned into a human pcDNA3.1 vector and transfected with Chinese hamster ovary cells (CHO) with a cAMP response element-regulated green fluorescent protein reporter gene. ) Screening for a stable expression of the β 2 receptor from the positive monoclonal cells obtained and a dose-dependent green fluorescence intensity of the cell line after stimulation by the agonist. This screening system was used to screen for β 2 -receptors from traditional Chinese medicines. Agonist-active compounds. Synephine was obtained from several Rutaceae herbs and higenamine was obtained from Asarum and Aconite. For the first time, their β 2 -agonism was demonstrated at the receptor level. Agent activity. This screening system is suitable for rapid screening of components with β 2 agonist activity from a large number of natural compounds.