Department of Neuroscience, Mayo Clinic College of Medicine Jacksonville, FL, USA.
Front Aging Neurosci. 2014 Oct 8;6:268. doi: 10.3389/fnagi.2014.00268. eCollection 2014.
Abnormal accumulation of filamentous α-synuclein (α-syn) in neurons, regarded as Lewy bodies (LBs), are a hallmark of Parkinson disease (PD). Although the exact mechanism(s) underlying LBs formation remains unknown, autophagy and ER stress response have emerged as two important pathways affecting α-syn aggregation. In present study we tested whether cells with the tetracycline-off inducible overexpression of α-syn and accumulating α-syn aggregates can benefit from autophagy activation elicited by nutrient deprivation (ND), since this approach was reported to effectively clear cellular polyglutamine aggregates. We found that nutrient deprivation of non-induced cells did not affect cell viability, but significantly activated autophagy reflected by increasing the level of autophagy marker LC3-II and autophagic flux and decrease of endogenous α-syn. Cells with induced α-syn expression alone displayed autophagy activation in an α-syn dose-dependent manner to reach a level comparable to that found in non-induced, nutrient deprived counterparts. Nutrient deprivation also activated autophagy further in α-syn induced cells, but the extent was decreased with increase of α-syn dose, indicating α-syn overexpression reduces the responsiveness of cells to nutrient deprivation. Moreover, the nutrient deprivation enhanced α-syn aggregations concomitant with significant increase of apoptosis as well as ER stress response, SREBP2 activation and cholesterolgenesis. Importantly, α-syn aggregate accumulation and other effects caused by nutrient deprivation were counteracted by knockdown of SREBP2, treatment with cholesterol lowering agent-lovastatin, or by GRP78 overexpression, which also caused decrease of SREBP2 activity. Similar results were obtained from studies of primary neurons with α-syn overexpression under nutrient deprivation. Together our findings suggested that down-regulation of SREBP2 activity might be a means to prevent α-syn aggregation in PD via reducing cholesterol levels.
异常积累丝状α-突触核蛋白(α-syn)在神经元中,被认为是路易体(LB),是帕金森病(PD)的标志。虽然 LB 形成的确切机制尚不清楚,但自噬和 ER 应激反应已成为影响α-syn 聚集的两个重要途径。在本研究中,我们测试了是否可以通过营养剥夺(ND)诱导的自噬激活来使过表达α-syn 并积累α-syn 聚集的细胞受益,因为据报道这种方法可以有效地清除细胞聚谷氨酰胺聚集物。我们发现,未诱导的细胞的营养剥夺不会影响细胞活力,但会显著激活自噬,表现为自噬标志物 LC3-II 和自噬流的增加以及内源性α-syn 的减少。单独表达α-syn 的细胞在α-syn 剂量依赖性方式下显示出自噬激活,达到与未诱导的、营养剥夺的对应物相当的水平。营养剥夺也进一步激活了α-syn 诱导的细胞中的自噬,但程度随着α-syn 剂量的增加而降低,表明α-syn 过表达降低了细胞对营养剥夺的反应性。此外,营养剥夺增强了α-syn 的聚集,同时伴随着凋亡以及 ER 应激反应、SREBP2 激活和胆固醇生成的显著增加。重要的是,营养剥夺引起的α-syn 聚集和其他效应被 SREBP2 敲低、胆固醇降低剂洛伐他汀处理或 GRP78 过表达所抵消,这也导致 SREBP2 活性降低。在营养剥夺下过表达α-syn 的原代神经元的研究中也获得了类似的结果。总之,我们的研究结果表明,下调 SREBP2 活性可能是通过降低胆固醇水平来预防 PD 中α-syn 聚集的一种方法。