College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, Heilongjiang, China.
College of Animal Science, Yangtze University, Jingzhou, 434025, Hubei, China.
Theriogenology. 2022 Jan 1;177:1-10. doi: 10.1016/j.theriogenology.2021.10.005. Epub 2021 Oct 8.
Chloroquine (CQ) could function as a lysosomotropic agent to inhibit the endolysosomal trafficking in the autophagy pathway, and is widely used on malarial, tumor and recently COVID-19. However, the effect of CQ treatment on porcine immature Sertoli cells (iSCs) remains unclear. Here we showed that CQ could reduce iSC viability in a dose-dependent manner. CQ treatment (20 μM) on iSCs for 36h could elevate oxidative stress, damage mitochondrial function and promote apoptosis, which could be partially rescued by melatonin (MT) (10 nM). Transcriptome profiling identified 1611 differentially expressed genes (DEGs) (776 up- and 835 down-regulated) (20 μM CQ vs. DMSO), mainly involved in MAPK cascade, cell proliferation/apoptosis, HIF-1, PI3K-Akt and lysosome signaling pathways. In contrast, only 467 (224 up- and 243 down-regulated) DEGs (CQ + MT vs. DMSO) could be found after MT (10 nM) addition, enriched in cell cycle, regulation of apoptotic process, lysosome and reproduction pathways. Therefore, the partial rescue effects of MT on CQ treatment were confirmed by multiple assays (cell viability, ROS level, mitochondrial function, apoptosis, and mRNA levels of selected genes). Collectively, CQ treatment could impair porcine iSC viability by deranging the signaling pathways related to apoptosis and autophagy, which could be partially rescued by MT supplementation.
氯喹（CQ）可作为溶酶体趋向性药物，抑制自噬途径中的内溶酶体转运，广泛用于疟疾、肿瘤，最近还用于 COVID-19。然而，CQ 处理对猪未成熟支持细胞（iSCs）的影响尚不清楚。本研究表明，CQ 可呈剂量依赖性降低 iSC 活力。CQ 处理（20 μM）iSCs 36 h 可增加氧化应激，损害线粒体功能并促进细胞凋亡，而褪黑素（MT）（10 nM）可部分挽救。转录组谱分析鉴定出 1611 个差异表达基因（DEGs）（776 个上调和 835 个下调）（20 μM CQ 与 DMSO 相比），主要涉及 MAPK 级联、细胞增殖/凋亡、HIF-1、PI3K-Akt 和溶酶体信号通路。相比之下，只有 467 个 DEGs（CQ+MT 与 DMSO 相比）（224 个上调和 243 个下调）在添加 MT（10 nM）后才能被发现，这些 DEGs 富集在细胞周期、凋亡过程调控、溶酶体和生殖途径中。因此，MT 对 CQ 处理的部分挽救作用通过多种检测（细胞活力、ROS 水平、线粒体功能、细胞凋亡和选定基因的 mRNA 水平）得到了证实。综上所述，CQ 处理通过扰乱与细胞凋亡和自噬相关的信号通路可损害猪 iSCs 的活力，而 MT 的补充可部分挽救这种损伤。
