Joint Laboratory of Modern Agricultural Technology International Cooperation, Ministry of Education, Jilin Agricultural University, Changchun 130118, China.
Key Lab of Animal Production, Product Quality and Security, Ministry of Education, Jilin Agricultural University, Changchun 130118, China.
Molecules. 2020 Mar 9;25(5):1230. doi: 10.3390/molecules25051230.
Melatonin influences physiological processes such as promoting proliferation and regulating cell development and function, and its effects on chicken Sertoli cells are unknown. Therefore, we investigated the effects of melatonin on cell proliferation and its underlying mechanisms in chicken Sertoli cells. Chicken Sertoli cells were exposed to varying melatonin concentrations (1, 10, 100, and 1000 nM), and the melatonin-induced effects on cell proliferation were measured by Cell Counting Kit 8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), real-time qPCR, and western blotting. We found that 1000 nM melatonin significantly ( < 0.05) promoted cell proliferation in chicken Sertoli cells. Furthermore, melatonin significantly ( < 0.05) increased the expression of inhibin alpha subunit (INHA), and the silencing of INHA reversed the melatonin-induced effects on Sertoli cell proliferation. We also found that melatonin activates the extracellular-regulated protein kinase (ERK) signaling pathway. To explore the role of the ERK signaling pathway in melatonin-induced cell proliferation, PD98059 (an inhibitor of EKR1/2) was used to pre-treat chicken Sertoli cells. The melatonin-induced proliferation of chicken Sertoli cells was reversed by PD98059, with decreased cell viability, weakened cell proliferation, and down-regulated expression of the proliferating cell nuclear antigen (PCNA), cyclin D1 (CCND1) and INHA. In summary, our results indicate that melatonin promotes the proliferation of chicken Sertoli cells by activating the ERK/inhibin alpha subunit signaling pathway.
褪黑素影响生理过程，如促进增殖和调节细胞发育和功能，但其对鸡睾丸支持细胞的影响尚不清楚。因此，我们研究了褪黑素对鸡睾丸支持细胞增殖的影响及其潜在机制。鸡睾丸支持细胞暴露于不同浓度的褪黑素（1、10、100 和 1000 nM），通过 Cell Counting Kit 8（CCK-8）、5-乙炔基-2'-脱氧尿苷（EdU）、实时 qPCR 和 Western blot 测定褪黑素对细胞增殖的影响。我们发现 1000 nM 褪黑素可显著（<0.05）促进鸡睾丸支持细胞增殖。此外，褪黑素显著（<0.05）增加了抑制素 alpha 亚基（INHA）的表达，而 INHA 的沉默逆转了褪黑素对睾丸支持细胞增殖的影响。我们还发现褪黑素激活细胞外调节蛋白激酶（ERK）信号通路。为了探讨 ERK 信号通路在褪黑素诱导细胞增殖中的作用，使用 PD98059（ERK1/2 的抑制剂）预先处理鸡睾丸支持细胞。PD98059 逆转了褪黑素诱导的鸡睾丸支持细胞增殖，细胞活力降低，增殖减弱，增殖细胞核抗原（PCNA）、细胞周期蛋白 D1（CCND1）和 INHA 的表达下调。综上所述，我们的结果表明，褪黑素通过激活 ERK/抑制素 alpha 亚基信号通路促进鸡睾丸支持细胞增殖。
