College of Life Science, Yantai University, Yantai, Shandong Province, PR China.
Center of Reproductive Medicine, Shengjing Hospital of China Medical University, Shenyang, Liaoning Province, PR China.
Folia Histochem Cytobiol. 2022;60(3):226-236. doi: 10.5603/FHC.a2022.0021. Epub 2022 Aug 5.
Peroxiredoxin 6 (Prdx6) is widely expressed in mammalian tissues. Our previous study demonstrated that Prdx6 was expressed in human epididymis, present in human seminal fluid, and in spermatozoa. The protective role of Prdx6 in maintaining the viability and DNA integrity of human spermatozoa was also detected. Here, we demonstrate the potential role and mechanism of Prdx6 in human epididymis epithelial cells (HEECs).
Western blotting was used to measure expression levels of key proteins in the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. The malonaldehyde (MDA) levels and antioxidant capacity in HEECs were detected with the commercial kits. Digital gene expression analysis (DGE) was used to identify gene expression patterns in control and Prdx6-interference HEECs. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to validate the DGE findings.
Compared to control HEECs, the expression levels of JAK1, STAT1, phosphorylated JAK1 and STAT1 were significantly increased, while the expression level of SOCS3 was significantly decreased in Prdx6-interference HEECs. The MDA level and total antioxidant capacity in Prdx6-interference HEECs were significantly increased and decreased compared to that of control, respectively. DGE analysis identified 589 up-regulated and 314 down-regulated genes (including Prdx6) in Prdx6-interference HEECs. Thirteen significantly different pathways were identified between the two groups, with the majority of genes belonging to the CCL, CXCL, IL, and IFIT family of proteins and were related to immunity. In particular, the expression levels of IL6, IL6ST, and eighteen IFN-related genes were significantly increased in Prdx6-interference HEECs compared to control HEECs.
We found that reduced Prdx6 expression induced higher ROS levels in HEECs, which resulted in the activation of the IL-6 receptor and IFNγ expression to induce the JAK1/STAT1 signaling pathway.
过氧化物酶 6（Prdx6）广泛表达于哺乳动物组织中。我们之前的研究表明，Prdx6 存在于人附睾组织中，存在于人精液中，并存在于精子中。还检测到 Prdx6 在维持人精子活力和 DNA 完整性方面的保护作用。在这里，我们证明了 Prdx6 在人附睾上皮细胞（HEECs）中的潜在作用和机制。
使用 Western blot 法测量 Janus 激酶/信号转导和转录激活因子（JAK/STAT）信号通路中的关键蛋白表达水平。使用商业试剂盒检测 HEECs 中的丙二醛（MDA）水平和抗氧化能力。使用数字基因表达分析（DGE）鉴定对照和 Prdx6 干扰 HEECs 中的基因表达模式。逆转录-聚合酶链反应（RT-PCR）用于验证 DGE 结果。
与对照 HEECs 相比，Prdx6 干扰 HEECs 中 JAK1、STAT1、磷酸化 JAK1 和 STAT1 的表达水平显著增加，而 SOCS3 的表达水平显著降低。Prdx6 干扰 HEECs 中的 MDA 水平和总抗氧化能力分别显著增加和减少。DGE 分析鉴定出 Prdx6 干扰 HEECs 中有 589 个上调和 314 个下调基因（包括 Prdx6）。两组间鉴定出 13 个显著不同的通路，其中大部分基因属于 CCL、CXCL、IL 和 IFIT 家族蛋白，与免疫相关。特别是，与对照 HEECs 相比，Prdx6 干扰 HEECs 中 IL6、IL6ST 和 18 个 IFN 相关基因的表达水平显著增加。
我们发现 Prdx6 表达降低导致 HEECs 中 ROS 水平升高，进而激活 IL-6 受体和 IFNγ 表达，诱导 JAK1/STAT1 信号通路。
