Reproductive Medicine Center, Jiangxi Maternal and Child Health Hospital, Nanchang, Jiangxi, China.
Department of Orthopaedics, Nanchang Hongdu Hospital of Traditional Chinese Medicine, Nanchang, Jiangxi, China.
Bioengineered. 2021 Dec;12(2):12123-12134. doi: 10.1080/21655979.2021.2003943.
The molecular mechanism that triggers polycystic ovary syndrome (PCOS) is mysterious. Abnormal development of ovarian granulosa cells (GCs) is one of the causes of PCOS. Herein, our study was carried out using RNA-seq to detect the different gene expression levels in ovarian GCs between three patients with PCOS and four normal controls. To verify the RNA-seq data, GCs from 22 patients with PCOS and 21 controls with normal ovulation were collected to perform the RT-PCR analysis. Hedgehog signaling pathway (Hh) members, Ihh and Ptch2 were abnormally highly expressed in the PCOS tissue (PT). The qPCR also indicated that the expression levels of Hh signaling pathway downstream members, Ptch1, Gli1, and Gli2 in the PT were significantly higher than those in the normal tissue (NT). Besides, the expression of TNF-α mRNA in PCOS patients was higher than that in the control group. Through the chromatin immunoprecipitation assay (ChIP), we found that the Gli1-IP-DNA enriched from the granular cells of PCOS patients was higher than that of the control group. Finally, the Hh signaling pathway inhibitor, cyclopamine, can decrease the apoptosis of PCOS ovarian granulosa cells. These results suggest that abnormal activation of Hh signaling pathway, especially Ihh signal, may have a profound influence on PCOS.
多囊卵巢综合征（PCOS）发病的分子机制尚不清楚，卵巢颗粒细胞（GCs）的异常发育是 PCOS 的原因之一。本研究采用 RNA-seq 技术检测了 3 例 PCOS 患者和 4 例正常对照患者卵巢 GCs 中的不同基因表达水平。为了验证 RNA-seq 数据，收集了 22 例 PCOS 患者和 21 例排卵正常的对照患者的 GCs 进行 RT-PCR 分析。Hedgehog 信号通路（Hh）成员 Ihh 和 Ptch2 在 PCOS 组织（PT）中异常高表达。qPCR 还表明，PT 中 Hh 信号通路下游成员 Ptch1、Gli1 和 Gli2 的表达水平明显高于 NT。此外，PCOS 患者的 TNF-α mRNA 表达高于对照组。通过染色质免疫沉淀分析（ChIP），我们发现来自 PCOS 患者颗粒细胞的 Gli1-IP-DNA 的富集高于对照组。最后，Hh 信号通路抑制剂 cyclopamine 可减少 PCOS 卵巢颗粒细胞的凋亡。这些结果表明，Hh 信号通路的异常激活，特别是 Ihh 信号，可能对 PCOS 有深远的影响。
