Wan Shan, Chen Qingqing, Xiang Yu, Sang Yimiao, Tang Minyue, Song Yang, Feng Guofang, Ye Bingru, Bai Long, Zhu Yimin
Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, 310002, China; Key Laboratory of Reproductive Genetics (Ministry of Education) and Women's Reproductive Health Laboratory of Zhejiang Province, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310002, China.
Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, 310002, China; Key Laboratory of Reproductive Genetics (Ministry of Education) and Women's Reproductive Health Laboratory of Zhejiang Province, Women's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, 310002, China.
Mol Cell Endocrinol. 2023 May 1;566-567:111891. doi: 10.1016/j.mce.2023.111891. Epub 2023 Feb 17.
A multitude of cytokines have been reported to participate in the folliculogenesis process in female. Interleukin-1 (IL-1), belonging to interleukin family, is originally identified as an important immune factor involved in inflammation response. Besides the immunity system, IL-1 is also expressed in reproductive system. However, the role of IL-1 in regulating ovarian follicle function remains to be elucidated. In the current study, using the primary human granulosa-lutein (hGL) and immortalized human granulosa-like tumor cell line (KGN) models, we demonstrated that both IL-1α and IL-1β increased prostaglandin E2 (PGE2) production via upregulating its cyclooxygenase (COX) enzyme COX-2 expression in human granulosa cells. Mechanistically, IL-1α and IL-1β treatment activated nuclear factor kappa B (NF-κB) signaling pathway. Using the specific siRNA to knock down endogenous gene expression, we found that the inhibition of p65 expression abolished IL-1α and IL-1β-induced upregulation of COX-2 expression whereas knockdown of p50 and p52 had no effect. Moreover, our results also showed that IL-1α and IL-1β promoted the nuclear translocation of p65. ChIP assay demonstrated the transcriptional regulation of p65 on COX-2 expression. Additionally, we also found that IL-1α and IL-1β could activate the extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway. The inhibition of ERK1/2 signaling pathway activation reversed IL-1α and IL-1β-induced upregulation of COX-2 expression. Our findings shed light on the cellular and molecular mechanisms by which IL-1 modulates the COX-2 expression through NF-κB/P65 and ERK1/2 signaling pathways in human granulosa cells.
据报道，多种细胞因子参与女性卵泡发生过程。白细胞介素-1（IL-1）属于白细胞介素家族，最初被鉴定为参与炎症反应的重要免疫因子。除免疫系统外，IL-1在生殖系统中也有表达。然而，IL-1在调节卵巢卵泡功能中的作用仍有待阐明。在本研究中，我们使用原代人颗粒黄体细胞（hGL）和永生化人颗粒样肿瘤细胞系（KGN）模型，证明IL-1α和IL-1β均可通过上调人颗粒细胞中环氧化酶（COX）-2的表达来增加前列腺素E2（PGE2）的产生。机制上，IL-1α和IL-1β处理激活了核因子κB（NF-κB）信号通路。使用特异性小干扰RNA（siRNA）敲低内源性基因表达，我们发现抑制p65表达可消除IL-1α和IL-1β诱导的COX-2表达上调，而敲低p50和p52则无影响。此外，我们的结果还表明，IL-1α和IL-1β促进了p65的核转位。染色质免疫沉淀（ChIP）分析证明了p65对COX-2表达的转录调控。此外，我们还发现IL-1α和IL-1β可激活细胞外信号调节激酶1/2（ERK1/2）信号通路。抑制ERK1/2信号通路激活可逆转IL-1α和IL-1β诱导的COX-2表达上调。我们的研究结果揭示了IL-1通过NF-κB/P65和ERK1/2信号通路调节人颗粒细胞中COX-2表达的细胞和分子机制。
