Molecular and Clinical Pharmacology, ICBM, Faculty of Medicine, University of Chile, Independencia, 1027, Santiago, Chile.
Instituto de Ciencias Biomédicas, Facultad de Ciencias de la Salud, Universidad Autónoma de Chile, Santiago, Chile.
Neurotox Res. 2019 Jan;35(1):255-259. doi: 10.1007/s12640-018-9953-8. Epub 2018 Sep 10.
Aminochrome has been reported to induce lysosomal dysfunction by inhibiting the vacuolar H-type ATPase localized in lysosome membrane. DT-diaphorase has been proposed to prevent aminochrome neurotoxicity but it is unknown whether this enzyme prevents aminochrome-induced lysosomal dysfunction. In the present study, we tested the protective role of DT-diaphorase in lysosomal dysfunction by generating a cell line (SH-SY5YsiNQ7) with a stable expression of a siRNA against DT-diaphorase with only 10% expression of mRNA enzyme. The cells differentiated with retinoic acid and 12-o-tetradecanoylphorbol-13-acetate show a significant increase in the expression of tyrosine hydroxylase, vesicular monoamine transporter-2, and dopamine transporter. The incubation of SH-SY5YsiNQ7 cells with 10 μM aminochrome resulted in a significant decrease of lysosome pH determined by using acridine orange, while aminochrome has no effect on SH-SY5Y cells. These results support the proposed protective role of DT-diaphorase against aminochrome-induced lysosomal dysfunction.
氨甲环酸已被报道通过抑制定位于溶酶体膜的液泡型 H+-ATP 酶来诱导溶酶体功能障碍。二氢硫辛酰胺脱氢酶已被提议预防氨甲环酸的神经毒性，但尚不清楚该酶是否能预防氨甲环酸诱导的溶酶体功能障碍。在本研究中，我们通过生成一个稳定表达针对二氢硫辛酰胺脱氢酶的 siRNA 的细胞系（SH-SY5YsiNQ7），该细胞系的 mRNA 酶表达仅为 10%，从而检测了二氢硫辛酰胺脱氢酶在溶酶体功能障碍中的保护作用。用维甲酸和 12-O-十四烷酰佛波醇-13-醋酸酯分化的细胞显示酪氨酸羟化酶、囊泡单胺转运体-2 和多巴胺转运体的表达显著增加。用吖啶橙测定时，10 μM 氨甲环酸孵育 SH-SY5YsiNQ7 细胞会导致溶酶体 pH 显著下降，而氨甲环酸对 SH-SY5Y 细胞没有影响。这些结果支持二氢硫辛酰胺脱氢酶对氨甲环酸诱导的溶酶体功能障碍的保护作用。
