de Siqueira Erlânia Alves, Magalhães Emanuel Paula, de Assis Albert Layo Costa, Sampaio Tiago Lima, Lima Danya Bandeira, Marinho Marcia Machado, Martins Alice Maria Costa, de Andrade Geanne Matos, de Barros Viana Glauce Socorro
Department of Physiology and Pharmacology, Federal University of Ceará (UFC), Fortaleza, Ceará, Brazil.
Department of Clinical and Toxicological Analysis, Federal University of Ceará (UFC), Fortaleza, Ceará, Brazil.
Neurochem Res. 2023 Jan;48(1):250-262. doi: 10.1007/s11064-022-03735-5. Epub 2022 Sep 6.
Parkinson's disease (PD) is characterized by dopaminergic cell loss in the substantia nigra, and PD brains show neuroinflammation, oxidative stress, and mitochondrial dysfunction. The study evaluated the neuroprotective activity of 1α,25-dihydroxy vitamin D3 (VD3), on the rotenone (ROT)-induced cytotoxicity in PC12 cells. The viability parameters were assessed by the MTT and flow cytometry, on cells treated or not with VD3 and/or ROT. Besides, ROS production, cell death, mitochondrial transmembrane potential, reduced GSH, superoxide accumulation, molecular docking (TH and Keap1-Nrf2), and TH, Nrf2, NF-kB, and VD3 receptor protein contents by western blot were evaluated. VD3 was shown to improve the viability of ROT-exposed cells. Cells exposed to ROT showed increased production of ROS and superoxide, which decreased after VD3. ROT decrease in the mitochondrial transmembrane potential was prevented, after VD3 treatment and, VD3 was shown to interact with tyrosine hydroxylase (TH) and Nrf2. While ROT decreased TH, Nrf2, and NF-kB expressions, these effects were reversed by VD3. In addition, VD3 also increased VD3 receptor protein contents and values went back to those of controls after ROT exposure. VD3 protects PC12 cells against ROT damage, by decreasing oxidative stress and improving mitochondrial function. One target seems to be the TH molecule and possibly an indirect Nrf2 activation could also justify its neuroprotective actions on this PC12 cell model of PD.
帕金森病（PD）的特征是黑质中多巴胺能细胞丢失，且PD患者的大脑表现出神经炎症、氧化应激和线粒体功能障碍。该研究评估了1α,25 - 二羟基维生素D3（VD3）对鱼藤酮（ROT）诱导的PC12细胞毒性的神经保护活性。通过MTT法和流式细胞术评估了用VD3和/或ROT处理或未处理的细胞的活力参数。此外，还通过蛋白质免疫印迹法评估了活性氧（ROS）生成、细胞死亡、线粒体跨膜电位、还原型谷胱甘肽（GSH）减少、超氧化物积累、分子对接（酪氨酸羟化酶（TH）和Keap1 - Nrf2）以及TH、Nrf2、核因子κB（NF - kB）和VD3受体蛋白含量。结果表明，VD3可提高ROT处理细胞的活力。暴露于ROT的细胞显示ROS和超氧化物生成增加，而VD3处理后这些物质减少。VD3处理可防止ROT引起的线粒体跨膜电位降低，且VD3显示与酪氨酸羟化酶（TH）和Nrf2相互作用。虽然ROT降低了TH、Nrf2和NF - kB的表达，但VD3可逆转这些作用。此外，VD3还增加了VD3受体蛋白含量，且ROT暴露后其值恢复至对照水平。VD3通过降低氧化应激和改善线粒体功能来保护PC12细胞免受ROT损伤。一个靶点似乎是TH分子，并且可能间接激活Nrf2也可以解释其在该PD的PC12细胞模型上的神经保护作用。
