RIKILT-Institute of Food Safety, Wageningen University and Research Centre, Akkermaalsbos 2, 6708 WB Wageningen, The Netherlands.
Anal Bioanal Chem. 2011 Aug;401(3):873-82. doi: 10.1007/s00216-011-5162-5. Epub 2011 Jun 18.
Sensitive and robust bioassays for glucocorticoids are very useful for the pharmaceutical industry, environmental scientists and veterinary control. Here, a recombinant yeast cell was constructed that expresses the human glucocorticoid receptor alpha and a green fluorescent reporter protein in response to glucocorticoids. Both the receptor construct and the reporter construct were stably integrated into the yeast genome. The correct and specific functioning of this yeast glucocorticoid bioassay was studied by exposures to cortisol and other related compounds and critically compared to a GR-CALUX bioassay based on a human bone cell. Although less sensitive, the new yeast glucocorticoid bioassay showed sensitivity towards all (gluco)corticoids tested, with the following order in relative potencies: budesonide >> corticosterone > dexamethasone > cortisol = betamethasone > prednisolone > aldosterone. Hormone representatives for other hormone nuclear receptors, like 17β-estradiol for the oestrogen receptor, 5α-dihydrotestosterone for the androgen receptor and progesterone for the progesterone receptor, showed no clear agonistic responses, whilst some polychlorinated biphenyls were clearly able to interfere with the GR activity.
用于检测糖皮质激素的灵敏且稳健的生物测定法对制药业、环境科学家和兽医控制非常有用。在此,构建了一种重组酵母细胞,该细胞可响应糖皮质激素表达人糖皮质激素受体 alpha 和绿色荧光报告蛋白。受体构建体和报告构建体都稳定地整合到酵母基因组中。通过暴露于皮质醇和其他相关化合物,对这种酵母糖皮质激素生物测定法的正确和特异性功能进行了研究,并与基于人骨细胞的 GR-CALUX 生物测定法进行了严格比较。尽管灵敏度较低,但新的酵母糖皮质激素生物测定法对所有(糖)皮质激素均表现出敏感性,相对效价顺序为:布地奈德>皮质酮>地塞米松>皮质醇=倍他米松>泼尼松龙>醛固酮。其他激素核受体的激素代表物,如雌激素受体的 17β-雌二醇、雄激素受体的 5α-二氢睾酮和孕激素受体的孕酮,没有表现出明显的激动反应,而一些多氯联苯显然能够干扰 GR 活性。