多囊卵巢综合征（PCOS）是一类以高雄激素血症为主要特征的内分泌代谢紊乱性疾病。高雄激素是导致卵泡闭锁、卵巢多囊样改变的主要因素，3β-羟甾脱氢酶亚型2（HSD3B2）是肾上腺-性腺雄激素合成的关键酶，其活性受多种转录因子调节。补肾活血化痰方（BHHR）是临床治疗PCOS的经验方，前期研究显示BHHR可显著降低PCOS高雄激素水平，全基因表达谱、GO分析等生物信息学技术检测提示BHHR可明显抑制HSD3B2基因表达。由此我们提出假说：BHHR可能通过影响HSD3B2基因上游调控元件上的关键转录因子的活性、抑制PCOS雄激素合成从而发挥治疗作用。因此本课题拟通过体外构建牛卵泡膜细胞的原代无血清培养体系，采用质粒构建、细胞转染、RNA干扰及EMSA等技术阐明调节PCOS HSD3B2基因表达的关键转录因子以及BHHR对PCOS HSD3B2基因转录的调控机制，为BHHR的临床推广应用奠定基础。

Polycystic ovary syndrome (PCOS), with hyperandrogenism as the main characteristics, is a kind of endocrine metabolic disorder. High level of androgen is the main factors of follicular atresia and morphological changes of ovary. 3β- hydroxysteroid dehydrogenase Subtype 2 (HSD3B2), one of the key enzymes of androgen synthesis of adrenal- the gonads, can be regulated by a variety of transcription factors. Bushen Huoxue Huatan Recipe (BHHR) is a commonly used prescription for the treatment of PCOS in clinic. Our early studies have shown that BHHR could significantly reduce the high level of androgen of PCOS. Besides, BHHR could obviously inhibit the expression of HSD3B2 detected by bioinformatics technologies including Genome-wide expression profile and GO analysis. Thus, we propose a hypothesis that the mechanisms of BHHR to treat PCOS might by affecting the transcription activity of HSD3B2 and inhibiting the synthesis of androgen. Based on the above researches, we plan to culture the bovine theca cells in vitro and explore the key transcription factors of HSD3B2 by using Plasmid construct, Cell transfection, RNAi, EMSA and other molecular biology techniques. This research will clarify the exact transcription factors regulated PCOS HSD3B2 gene and the mechanisms of BHHR on the transcription factors. Therefore, to provide theoretical basis for clinical application of BHHR.