The Effect of Cellular Coenzyme Q10 Deficiency on Lysosomal Acidification
The Effect of Cellular Coenzyme Q10 Deficiency on Lysosomal Acidification
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DOI：
10.3390/jcm9061923
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发表时间：
2020-06-01
影响因子：
3.9
通讯作者：
Hargreaves, Iain
中科院分区：
文献类型：
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作者：
Heaton, Robert A.;Heales, Simon;Hargreaves, Iain
辅酶 Q(10)(CoQ(10)) 缺乏症是目前唯一可治疗的线粒体疾病，然而，人们对其如何影响其他细胞器知之甚少。已发现溶酶体的膜上有大量 CoQ(10)；此外，有人认为它在溶酶体腔的正常酸化中发挥作用。因此，在本研究中，我们评估了 CoQ(10) 缺乏对溶酶体酸化的影响。为了研究这一点，通过用对氨基苯甲酸（PABA）处理SH-SY5Y细胞建立了CoQ（10）缺陷的神经元细胞模型。该方法通过竞争性抑制 CoQ(10) 生物合成途径酶 CoQ(2) 发挥作用。单次 1 mM（5 天）PABA 处理可导致细胞 CoQ(10) 下降高达 58%（p < 0.05）。结果发现，这导致用于测量溶酶体 pH 值的 LysoSensor (23%) 和 LysoTracker (35%) 探针的荧光显着降低 (p< 0.05)。结果发现，随后用 CoQ(10)（5 μM，3 天）处理能够恢复细胞 CoQ(10) 浓度 (p< 0.005)，这与两个探针的荧光增加至对照的 90% 左右 (p< 0.05) 相关，表明溶酶体 pH 值恢复。这项研究深入了解了溶酶体 pH 值与细胞 CoQ(10) 状态之间的关联，以及这种类异戊二烯状态的缺陷可能导致溶酶体酸化受损的可能性。
Coenzyme Q(10)(CoQ(10)) deficiency currently represents the only treatable mitochondrial disorder, however, little is known about how it may affect other organelles. The lysosome has been found to have a large concentration of CoQ(10)localised at its membrane; additionally, it has been suggested that it plays a role in the normal acidification of the lysosomal lumen. As a result, in this study we assessed the effect of CoQ(10)deficiency on lysosomal acidification. In order to investigate this, a neuronal cell model of CoQ(10)deficiency was established via the treatment of SH-SY5Y cells with para-aminobenzoic acid (PABA). This method works through the competitive inhibition of the CoQ(10)biosynthetic pathway enzyme, CoQ(2). A single 1 mM (5 days) treatment with PABA resulted in a decrease of up to 58% in cellular CoQ(10)(p< 0.05). It was found that this resulted in a significant decrease in fluorescence of both the LysoSensor (23%) and LysoTracker (35%) probes used to measure lysosomal pH (p< 0.05). It was found that subsequent treatment with CoQ(10)(5 mu M, 3 days) was able to restore cellular CoQ(10)concentration (p< 0.005), which was associated with an increase in fluorescence from both probes to around 90% of controls (p< 0.05), suggesting a restoration of lysosomal pH. This study provides insights into the association between lysosomal pH and cellular CoQ(10)status and the possibility that a deficit in the status of this isoprenoid may result in an impairment of lysosomal acidification.