METHODS: DHEA was used to construct the PCOS rat model. After intervention with quercetin, metformin, and AR, the estrous cycle, ovarian and uterine weight of rats were measured. The morphological changes of ovarian and uterine were detected by hematoxylin-eosin staining (HE staining). Luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured by Enzyme linked immunosorbent assay (ELISA). Immunohistochemical detection of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-alpha (TNF-α), B-cell lymphoma-2 (BCL-2), BCL2-Associated X (Bax) and AR expression in ovarian. Determination of the expression of CNP and NPR2 mRNA by qRT-PCR. Chromatin immunocoprecipitation (ChIP) was used to detect the ability of AR to bind to CNP or NPR2 promoter.
RESULTS: The results showed that quercetin could significantly reduce the expression of Testosterone (T) , Estradiol (E2) , LH, Bax, IL-1β, IL-6 and TNF-α, increase the expression of FSH and Bcl-2, inhibit the expression of AR, regulate the expression of CNP / NPR2 gene and protein by affecting the combination of AR with the specific sequence of CNP and NPR2 gene promoters, restore the maturation of oocyte and ovulation.
CONCLUSIONS: The results suggest that quercetin can alleviate the hormone, metabolic and ovulatory aberrations caused by PCOS, and provide experimental basis for the clinical application of quercetin in PCOS.
方法:采用DHEA构建PCOS大鼠模型。槲皮素干预后,二甲双胍,AR,发情周期,测量大鼠卵巢和子宫重量。采用苏木精-伊红染色(HE染色)检测卵巢和子宫的形态学变化。采用酶联免疫吸附试验(ELISA)检测促黄体生成素(LH)和卵泡刺激素(FSH)。免疫组织化学检测白细胞介素-6(IL-6),白细胞介素-1β(IL-1β),肿瘤坏死因子-α(TNF-α),B细胞淋巴瘤-2(BCL-2),BCL2相关X(Bax)和AR在卵巢中的表达。qRT-PCR测定CNP和NPR2mRNA的表达。染色质免疫沉淀(ChIP)用于检测AR与CNP或NPR2启动子结合的能力。
结果:结果表明槲皮素能显著降低睾酮(T)的表达,雌二醇(E2),LH,Bax,IL-1β,IL-6和TNF-α,增加FSH和Bcl-2的表达,抑制AR的表达,通过影响AR与CNP和NPR2基因启动子的特异性序列的结合来调控CNP/NPR2基因和蛋白的表达,恢复卵母细胞的成熟和排卵。
结论:结果表明槲皮素可以缓解激素,由PCOS引起的代谢和排卵异常,为槲皮素在PCOS中的临床应用提供实验依据。