Core clock gene Bmal1 deprivation impairs steroidogenesis in mice luteinized follicle cells.
Core clock gene Bmal1 deprivation impairs steroidogenesis in mice luteinized follicle cells.
复制标题
核心时钟基因 Bmal1 剥夺会损害小鼠黄素化卵泡细胞的类固醇生成
DOI：
10.1530/rep-20-0340
复制
发表时间：
2020-12
期刊：
影响因子：
--
通讯作者：
Xu Y
中科院分区：
文献类型：
--
作者：
Wang Y;Chen M;Xu J;Liu X;Duan Y;Zhou C;Xu Y
黄体化是黄体形成的过程，是卵泡细胞转化的一种方式，是类固醇合成的一个改变过程。Bmal 1基因作为核心时钟基因，参与调控排卵过程及随后的黄体功能。到目前为止，Bmal 1在黄体化中的潜在作用仍不清楚。为了探讨Bmal 1在黄体类固醇生成中的独特作用及其潜在途径，我们研究了Bmal 1基因敲除雌性小鼠的黄体激素合成谱。结果显示，卵巢癌患者黄体激素合成明显受损，PI 3 K/NFκB通路磷酸化明显激活。然后，在体外培养的细胞中验证结果，包括分离的Bmal 1干扰颗粒细胞（GC）和卵泡膜细胞（TC）。在体外黄体化过程中，培养上清中激素水平和类固醇生成相关基因（GC中星星、Hsd 3 β2、cyp 19 a1，TC中Lhcgr、星星、Hsd 3 β2、cyp 17 a1）的mRNA表达相互降低，而PI 3 K/NFκB的磷酸化水平升高。PI 3 K特异性抑制剂LY 294002干预后，Lhcgr和Hsd 3 β2的mRNA表达部分恢复，雄烯二酮和T合成显著增加。进一步研究表明，BMAL 1与NFκB p65（RelA）直接负性相互作用，RelA被认为是BMAL 1调控的PI 3 K信号转导的介导因子。综上所述，我们证实了Bmal 1在黄体类固醇生成中的新作用，通过与RelA介导的PI 3 K/NFκB通路的负相互作用实现。
Luteinization is the event of corpus luteum formation, a way of follicle cells transformation and a process of steroidogenesis alteration. As the core clock gene, Bmal1 was involved in the regulation of ovulation process and luteal function afterwards. Till now, the underlying roles of luteinization played by Bmal1 remain unknown. To explore the unique role of Bmal1 in luteal steroidogenesis and its underlying pathway, we investigated the luteal hormone synthesis profile in Bmal1 knockout female mice. We found that luteal hormone synthesis was notably impaired, and phosphorylation of PI3K/NfκB pathway was significantly activated. Then, the results were verified in in vitro cultured cells, including isolated Bmal1 interference granulosa cells (GCs) and theca cells (TCs), respectively. Hormones levels of supernatant culture media and mRNA expressions of steroidogenesis-associated genes (star, Hsd3β2, cyp19a1 in GCs, Lhcgr, star, Hsd3β2, cyp17a1 in TCs) were mutually decreased, while the phosphorylation of PI3K/NfκB was promoted during in vitro luteinization. After PI3K specific-inhibitor LY294002 intervention, mRNA expressions of Lhcgr and Hsd3β2 were partially rescued in Bmal1 interference TCs, together with significantly increased androstenedione and T synthesis. Further exploration in TCs demonstrated BMAL1 interacted directly but negatively with NfκB p65 (RelA), a subunit which was supposed as a mediator in Bmal1-governed PI3K signaling regulation. Taken together, we verified the novel role of Bmal1 in luteal steroidogenesis, achieving by negative interplay with RelA-mediated PI3K/NfκB pathway.