糖皮质激素（GC）的非基因组作用在机体生理和病理过程中具有重要的生物学意义。我们前期研究发现，GC与垂体ACTH细胞的细胞膜GR结合，快速抑制垂体细胞ACTH的分泌，属于非基因组作用，但其分子机制尚不明了。本项目（1）采用co-IP和质谱技术，确定介导GC非基因组作用的细胞膜蛋白谱，采用生物信息学方法、蛋白免疫印迹和细胞免疫荧光等技术，确定并验证候选蛋白CSN6；敲减CSN6，检测其对GR转运的影响；（2）采用磷酸化抗体芯片，确定调控GC非基因组作用的磷酸化信号通路；敲减CSN6，检测蛋白磷酸化水平的变化；抑制磷酸化信号通路对GR转运的影响；（3）采用大鼠垂体细胞灌注系统，验证CSN6和磷酸化信号通路与GC对ACTH分泌快速抑制作用的相关性。本项目的实施，对阐明GC非基因组作用的分子机制具有重要的学术意义，也将为皮质醇增多症等综合征的临床治疗提供理论基础。

The non-genomic action of glucocorticoids plays an important role in physiological and pathological processes. Our previous data showed that the rapid inhibitory effect of glucocorticoids on pituitary adreno-cortico-tropic-hormone (ACTH) secretion was a non-genomic action mediated by membrane glucocorticoid receptor (mGR), but the molecular mechanism was still unclear. In the present study, we will employ co-immunoprecipitation and mass spectrometry to screen and identify the key membrane protein profiling mediating the fast action. We will also use bio-informatics analysis, Western blot and immunefluorescence to identify and verify the candidate protein CSN6 (Constitutive photomorphogenesis 9 signalosome subunit 6). Then, we will knockdown CSN6 by shRNA transfection, and the GR trafficking in the cells transfected with CSN6 shRNA. To explore the mechanism that regulating the non-genomic action of glucocorticoids by CSN6, we will use protein phosphorylation arrays to set up a phosphorylated protein profiling. We will check the fold change of protein phosphorylation level, after CSN6 was knock down. And we will also conduct experiments to check the inhibitory effect of the phosphorylation signaling pathway on GR trafficking. Finally, we will apply the pituitary cell perifusion system to verify the relationship between the candidate proteins as well as the phosphorylation signaling pathway with the rapid inhibitory effects of glucocorticoids on pituitary ACTH secretion. The implementation of this project will have a critical academic significance on clarifying the non-genomic action of glucocorticoids, and then provide the theoretical basis for clinical treatment of Cushing syndrome.