Lithium facilitates removal of misfolded proteins and attenuated faulty interaction between mutant SOD1 and p-CREB (Ser133) through enhanced autophagy in mutant hSOD1(G93A) transfected neuronal cell lines
Lithium facilitates removal of misfolded proteins and attenuated faulty interaction between mutant SOD1 and p-CREB (Ser133) through enhanced autophagy in mutant hSOD1(G93A) transfected neuronal cell lines
复制标题
锂通过增强突变型 hSOD1(G93A) 转染神经元细胞系中的自噬，促进错误折叠蛋白的去除并减弱突变型 SOD1 和 p-CREB (Ser133) 之间的错误相互作用
DOI：
10.1007/s11033-019-05071-4
复制
发表时间：
2019
影响因子：
2.8
通讯作者：
Feng Honglin
中科院分区：
文献类型：
--
作者：
Yin Xiang;Wang Shuyu;Wang Xudong;Yang Yueqing;Jiang Hongquan;Wang Tianhang;Wang Ying;Zhang Chunting;Feng Honglin
蛋白质的异常聚集和沉积是ALS的重要病理特征，可能与细胞自噬功能失调有关。在本研究中，我们发现，与wtSOD 1细胞相比，血清饥饿处理导致mutSOD 1细胞的凋亡率显著升高;锂处理对mutSOD 1细胞具有保护作用，减少GFP标记的突变SOD 1蛋白聚集体的沉积;而Baf或3-MA预处理（自噬抑制剂）阻断了锂对突变SOD 1细胞的保护，并诱导增加的GFP标记的突变SOD 1蛋白聚集。此外，Western印迹结果显示，锂处理导致mutSOD 1细胞的Triton X-100可溶性和Triton X-100不溶性部分中的突变hSOD 1蛋白水平降低。此外，在mutSOD 1细胞中，突变SOD 1蛋白的聚集体与p-CREB（Ser 133）（转录因子）的不适当结合被证明;而锂处理减弱了这种错误的相互作用。总之，我们的结果表明，在mutSOD 1细胞中，mutSOD 1蛋白聚集体与异常的自噬调节有关。锂处理可以诱导自噬和增强蛋白聚集体的清除，进一步对mutSOD 1细胞发挥保护作用。更重要的是，我们发现了mutSOD 1蛋白聚集体的另一个独特的病理作用，即与p-CREB（Ser 133）的异常结合，p-CREB（Ser 133）是一种重要的转录因子，可能在PI 3 K-Akt-CREB-AEG-1信号通路中起关键作用。
Abnormally protein aggregation and deposition are key pathological features of ALS, which may related with dysfunctional cellular autophagy. In the current study, we found that, compared with wtSOD1 cells, serum starvation treatment resulted in significant higher percentage of apoptosis in mutSOD1 cells; Lithium treatment exerted protection for those mutSOD1 cells, with decreased GFP-tagged mutant SOD1 protein aggregates deposition; Whereas, pre-treatment with Baf or 3-MA (autophagy inhibitors) blocked protection of lithium for mutant SOD1 cells, and induced increased GFP-tagged mutant SOD1 protein aggregation. Further, Western blots results showed that lithium treatment led to decrease of mutant hSOD1 protein levels in both Triton X-100 soluble and Triton X-100 insoluble fraction of mutSOD1 cells. Besides, improper binding of mutant SOD1 proteins’ aggregates with p-CREB (Ser133) (transcription factor) in mutSOD1 cells were demonstrated; whereas lithium treatment attenuated this fault interaction. In conclusion, our results showed that, in mutSOD1 cells, mutSOD1 protein aggregates were related with abnormal autophagic regulation. Lithium treatment could induce autophagy and enhance clearance of protein aggregates, further exerting protection on mutSOD1 cells. More importantly, we uncovered another distinct pathological role of mutSOD1 protein aggregates, that is abnormal binding with p-CREB (Ser133), an important transcription factor, which may play crucial role in the PI3K-Akt-CREB-AEG-1 signaling pathway.