补肾活血化痰方调控多囊卵巢综合征3β-羟甾脱氢酶亚型2基因转录的机制研究

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.

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。

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。本项目在前期研究工作基础之上，1）开展动物实验，诱导PCOS高雄激素血症大鼠模型，研究发现BHHR可降低PCOS模型大鼠血清高雄激素水平，改善卵巢多囊样状态，对PCOS模型大鼠有显著的治疗作用，其作用机制可能与抑制卵泡膜细胞雄激素合成关键酶HSD3B2基因表达相关；2）通过细胞实验，采用质粒构建、细胞转染、RNA干扰等方法研究发现，BHHR可能是通过调节HSD3B2基因上游调控元件上位于-325~527位点的转录因子GATA-1的转录活性进而抑制PCOS雄激素合成；3）开展临床试验，采用前瞻性、随机、对照临床试验研究方法研究发现BHHR可显著调节PCOS患者紊乱的内分泌水平、改善患者明显患者的临床症状及中医症候，促进排卵有利于妊娠，其临床效果显著。