Marine Science Institute, The University of Texas at Austin, 750 Channel View Dr, Port Aransas, TX, 78373, USA.
J Mol Neurosci. 2022 Oct;72(10):2067-2080. doi: 10.1007/s12031-022-02057-z. Epub 2022 Aug 17.
We recently showed that membrane progesterone receptor α (mPRα/PAQR7) promotes pro-regenerative effects in Schwann cell-like adipose stem cells (SCL-ASC), an alternative model to Schwann cells for the promotion of peripheral nerve regeneration. In this study, we investigated how mPRα activation with the mPR-specific agonist Org OD 02-0 in SCL-ASC affected regenerative parameters in two neuronal cell lines, IMR-32 and SH-SY-5Y. In a series of conditioned medium experiments, we found that mPR activation of SCL-ASC led to increased neurite outgrowth, protection from cell death and increased expression of peripheral nerve regeneration markers (CREB3, ATF3, GAP43) in neuronal cell lines. These effects were stronger than the ones observed with the conditioned medium from untreated SCL-ASC. The addition of Org OD 02-0 to the untreated cell medium mimicked the effects of mPR activation of SCL-ASC on cell death, but not on neurite outgrowth. Therefore, the effect of Org OD 02-0 on neurite outgrowth is SCL-ASC-dependent, while its effect on cell survivability is likely due to the direct activation of mPRs on neuronal cells. SCL-ASC transfection with mPRα siRNA showed that this isoform is responsible for the beneficial effect on neurite outgrowth. Further experiments showed that SCL-ASC-dependent outcomes likely involved the release of BDNF and IGF-2 from these cells. The beneficial mPRα effect on neurite outgrowth was confirmed in co-culture conditions. These findings strengthen the hypothesis that mPRα could play a pro-regenerative role in SCL-ASC and be a therapeutic target for the promotion of peripheral nerve regeneration.
我们最近表明,膜孕激素受体α(mPRα/PAQR7)促进类雪旺细胞脂肪干细胞(SCL-ASC)的促再生作用,这是促进周围神经再生的雪旺细胞替代模型。在这项研究中,我们研究了 SCL-ASC 中 mPR 特异性激动剂 Org OD 02-0 的 mPR 激活如何影响两种神经元细胞系 IMR-32 和 SH-SY-5Y 的再生参数。在一系列条件培养基实验中,我们发现 SCL-ASC 的 mPR 激活导致神经元细胞系中的神经突生长增加、细胞死亡保护和周围神经再生标志物(CREB3、ATF3、GAP43)的表达增加。这些作用强于未经处理的 SCL-ASC 条件培养基观察到的作用。在未经处理的细胞培养基中添加 Org OD 02-0 模拟了 SCL-ASC 对细胞死亡的 mPR 激活作用,但对神经突生长没有作用。因此,Org OD 02-0 对神经突生长的作用是 SCL-ASC 依赖性的,而其对细胞存活的作用可能是由于 mPR 在神经元细胞上的直接激活。SCL-ASC 转染 mPRα siRNA 表明该同工型负责对神经突生长产生有益影响。进一步的实验表明,SCL-ASC 依赖性结果可能涉及这些细胞释放 BDNF 和 IGF-2。在共培养条件下证实了 mPRα 对神经突生长的有益作用。这些发现加强了 mPRα 在 SCL-ASC 中可能发挥促再生作用并且是促进周围神经再生的治疗靶点的假设。