Department of Anesthesiology, Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China.
Department of Anesthesiology and Pain Research Center, Jiaxing University Affiliated Hospital, The First Hospital of Jiaxing, Jiaxing, China.
J Cell Mol Med. 2020 Dec;24(24):14184-14194. doi: 10.1111/jcmm.16029. Epub 2020 Oct 27.
As a cis-acting non-depolarizing neuromuscular blocker through a nicotinic acetylcholine receptor (nAChR), cisatracurium (CAC) is widely used in anaesthesia and intensive care units. nAChR may be present on Leydig cells to mediate the action of CAC. Here, by Western blotting, immunohistochemistry and immunofluorescence, we identified that CHRNA4 (a subunit of nAChR) exists only on rat adult Leydig cells. We studied the effect of CAC on the synthesis of testosterone in rat adult Leydig cells and mouse MLTC-1 tumour cells. Rat Leydig cells and MLTC-1 cells were treated with CAC (5, 10 and 50 μmol/L) or nAChR agonists (50 μmol/L nicotine or 50 μmol/L lobeline) for 12 hours, respectively. We found that CAC significantly increased testosterone output in rat Leydig cells and mouse MLTC-1 cells at 5 μmol/L and higher concentrations. However, nicotine and lobeline inhibited testosterone synthesis. CAC increased intracellular cAMP levels, and nicotine and lobeline reversed this change in rat Leydig cells. CAC may increase testosterone synthesis in rat Leydig cells and mouse MLTC-1 cells by up-regulating the expression of Lhcgr and Star. Up-regulation of Scarb1 and Hsd3b1 expression by CAC was also observed in rat Leydig cells. In addition to cAMP signal transduction, CAC can induce ERK1/2 phosphorylation in rat Leydig cells. In conclusion, CAC binds to nAChR on Leydig cells, and activates cAMP and ERK1/2 phosphorylation, thereby up-regulating the expression of key genes and proteins in the steroidogenic cascade, resulting in increased testosterone synthesis in Leydig cells.
顺式阿曲库铵（cisatracurium，CAC）作为一种具有烟碱型乙酰胆碱受体（nAChR）活性的非去极化神经肌肉阻滞剂，被广泛应用于麻醉学和重症监护病房。nAChR 可能存在于莱迪希细胞上，以介导 CAC 的作用。在这里，我们通过 Western blot、免疫组织化学和免疫荧光技术鉴定，仅在成年大鼠莱迪希细胞上存在 CHRNA4（nAChR 的一个亚基）。我们研究了 CAC 对成年大鼠莱迪希细胞和小鼠 MLTC-1 肿瘤细胞中睾酮合成的影响。分别用 CAC（5、10 和 50 μmol/L）或 nAChR 激动剂（50 μmol/L 尼古丁或 50 μmol/L 洛贝林）处理大鼠莱迪希细胞和 MLTC-1 细胞 12 小时。我们发现 CAC 在 5 μmol/L 及更高浓度时可显著增加大鼠莱迪希细胞和小鼠 MLTC-1 细胞中睾酮的分泌。然而，尼古丁和洛贝林抑制了睾酮的合成。CAC 增加了细胞内 cAMP 水平，而尼古丁和洛贝林逆转了这一变化。CAC 可能通过上调 Lhcgr 和 Star 的表达来增加大鼠莱迪希细胞和小鼠 MLTC-1 细胞中睾酮的合成。在大鼠莱迪希细胞中，还观察到 CAC 上调了 Scarb1 和 Hsd3b1 的表达。除了 cAMP 信号转导外，CAC 还可以诱导大鼠莱迪希细胞中 ERK1/2 的磷酸化。总之，CAC 与莱迪希细胞上的 nAChR 结合，激活 cAMP 和 ERK1/2 磷酸化，从而上调类固醇生成级联中的关键基因和蛋白的表达，导致莱迪希细胞中睾酮的合成增加。
