Department of Physiology, Chonnam National University Medical School, Hwasun 58128, Jeollanam-do, Korea.
Int J Mol Sci. 2021 Feb 26;22(5):2322. doi: 10.3390/ijms22052322.
Parkinson's disease (PD) is an age-related neurodegenerative disease (NDD) characterized by the degenerative loss of dopaminergic neurons in the substantia nigra along with aggregation of α-synuclein (α-syn). Neurogenic differentiation of human adipose-derived stem cells (NI-hADSCs) by supplementary factors for 14 days activates different biological signaling pathways. In this study, we evaluated the therapeutic role of NI-hADSC-conditioned medium (NI-hADSC-CM) in rotenone (ROT)-induced toxicity in SH-SY5Y cells. Increasing concentrations of ROT led to decreased cell survival at 24 and 48 h in a dose- and time-dependent manner. Treatment of NI-hADSC-CM (50% dilution in DMEM) against ROT (0.5 μM) significantly increased the cell survival. ROT toxicity decreased the expression of tyrosine hydroxylase (TH). Western blot analysis of the Triton X-100-soluble fraction revealed that ROT significantly decreased the oligomeric, dimeric, and monomeric phosphorylated Serine129 (p-S129) α-syn, as well as the total monomeric α-syn expression levels. ROT toxicity increased the oligomeric, but decreased the dimeric and monomeric p-S129 α-syn expression levels. Total α-syn expression (in all forms) was increased in the Triton X-100-insoluble fraction, compared to the control. NI-hADSC-CM treatment enhanced the TH expression, stabilized α-syn monomers, reduced the levels of toxic insoluble p-S129 α-syn, improved the expression of neuronal functional proteins, regulated the Bax/Bcl-2 ratio, and upregulated the expression of pro-caspases, along with PARP-1 inactivation. Moreover, hADSC-CM treatment decreased the cell numbers and have no effect against ROT toxicity on SH-SY5Y cells. The therapeutic effects of NI-hADSC-CM was higher than the beneficial effects of hADSC-CM on cellular signaling. From these results, we conclude that NI-hADSC-CM exerts neuroregenerative effects on ROT-induced PD-like impairments in SH-SY5Y cells.
帕金森病（PD）是一种与年龄相关的神经退行性疾病（NDD），其特征是黑质中多巴胺能神经元的退行性丧失，以及α-突触核蛋白（α-syn）的聚集。通过补充因子对人脂肪源性干细胞（NI-hADSCs）进行神经发生分化 14 天，会激活不同的生物学信号通路。在这项研究中，我们评估了 NI-hADSC 条件培养基（NI-hADSC-CM）在鱼藤酮（ROT）诱导的 SH-SY5Y 细胞毒性中的治疗作用。随着浓度的增加，ROT 在 24 和 48 小时时以剂量和时间依赖的方式导致细胞存活率降低。NI-hADSC-CM（在 DMEM 中稀释 50%）对 ROT（0.5 μM）的处理显著增加了细胞存活率。ROT 毒性降低了酪氨酸羟化酶（TH）的表达。Triton X-100 可溶性部分的 Western blot 分析表明，ROT 显著降低了寡聚、二聚和单体磷酸化丝氨酸 129（p-S129）α-syn 的表达，以及总单体α-syn 的表达水平。ROT 毒性增加了寡聚体，但降低了二聚体和单体 p-S129 α-syn 的表达水平。与对照相比，Triton X-100 不溶性部分的总α-syn 表达（所有形式）增加。NI-hADSC-CM 处理增强了 TH 的表达，稳定了α-syn 单体，降低了有毒不溶性 p-S129 α-syn 的水平，改善了神经元功能蛋白的表达，调节了 Bax/Bcl-2 比值，并上调了前半胱氨酸蛋白酶的表达，同时使 PARP-1 失活。此外，hADSC-CM 处理减少了细胞数量，并且对 SH-SY5Y 细胞的 ROT 毒性没有影响。NI-hADSC-CM 的治疗效果高于 hADSC-CM 对细胞信号的有益影响。从这些结果中，我们得出结论，NI-hADSC-CM 对 ROT 诱导的 SH-SY5Y 细胞帕金森病样损伤具有神经再生作用。
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