Previous studies have confirmed that the beclin 1 complex plays a key role in the initial stage of autophagy and deregulated autophagy might involve in amyotrophic lateral sclerosis. However, the mechanism underlying ...Previous studies have confirmed that the beclin 1 complex plays a key role in the initial stage of autophagy and deregulated autophagy might involve in amyotrophic lateral sclerosis. However, the mechanism underlying altered autophagy associated with the beclin 1 complex remains un- clear. In this study, we transfected the Cu/Zn superoxide dismutase 1 G93A mutant protein into the motor neuron-like cell line NSC34 cultured in vitro. Western blotting and co-immunopre- cipitation showed that the Cu/Zn superoxide dismutase 1 G93A mutant enhanced the turnover of autophagic marker microtubule-associated protein light chain 3II （LC3Ⅱ） and stimulated the conversion of EGFP-LC3Ⅰ to EGFP-LC3Ⅱ, but had little influence on the binding capacity of the autophagy modulators ATG14L, rubicon, UVRAG, and hVps34 to beclin 1 during auto- phagosome formation. These results suggest that the amyotrophic lateral sclerosis-linked Cu/Zn superoxide dismutase I G93A mutant can upregulate autophagic activity in NSC34 cells, but that this does not markedly affect beclin 1 complex components.展开更多
The expression of cytosolic phospholipase A2 (cPLA2) expression is up-regulated in animal model of ALS and in patients with familial amyotrophic lateral sclerosis (fALS). Inhibition of cyclooxygenase 2 (COX2), which i...The expression of cytosolic phospholipase A2 (cPLA2) expression is up-regulated in animal model of ALS and in patients with familial amyotrophic lateral sclerosis (fALS). Inhibition of cyclooxygenase 2 (COX2), which is a downstream enzyme of cPLA2, ameliorates the impairment of motor function in the ALS model mice. Therefore, the arachidonic acid cascade, including the cPLA2-COX2 pathway, is an important therapeutic target of ALS. The current study was designed to investigate the potential of AK106-001616, an inhibitor of cPLA2, in protection of motor neuron cell death induced by mutant superoxide dismutase (SOD1<sup>G93A</sup>). AK106-001616 (1 - 10 μM) protected NSC34 cells (mouse motor neuron like cells) against SOD1<sup>G93A</sup>-induced motor neuron cell death. Furthermore, aspirin, an inhibitor of COX1/2, reduced the SOD1<sup>G93A</sup>-induced motor neuron cell death at a concentration that inhibited COX2. Celecoxib, a selective COX2 inhibitor, also reduced the SOD1<sup>G93A</sup>-induced motor neuron cell death. These results suggest that the arachidonic acid cascade is important for SOD1<sup>G93A</sup>-induced motor neuron cell death and AK106-001616 has a potent neuroprotective effect against it. AK106-001616 may be a useful therapeutic agent against SOD1<sup>G93A</sup>-induced ALS.展开更多
目的:观察Notch1在肌萎缩侧索硬化症(ALS)转基因鼠动物模型和细胞模型中的表达情况。方法:应用免疫荧光、免疫印迹、RT-PCR,检测Notch1在95、108、122 d ALS转基因鼠脊髓中的表达变化;检测转染pEGFP-wt-SOD1和pEGFPG93A-SOD1的NSC34细...目的:观察Notch1在肌萎缩侧索硬化症(ALS)转基因鼠动物模型和细胞模型中的表达情况。方法:应用免疫荧光、免疫印迹、RT-PCR,检测Notch1在95、108、122 d ALS转基因鼠脊髓中的表达变化;检测转染pEGFP-wt-SOD1和pEGFPG93A-SOD1的NSC34细胞模型中Notch1的表达变化。结果:Notch1可与β-tubulinⅢ共表达,与GFAP无明显共表达。较同窝野生型鼠,Notch1于蛋白水平和mRNA水平上的表达在95 d ALS转基因鼠脊髓中无明显变化,在108 d和122 d ALS转基因鼠脊髓中明显升高;与转染pEGFP-wt-SOD的NSC34细胞相比,转染pEGFP-G93A-SOD1的NSC34细胞中Notch1蛋白和mRNA表达增多。结论:Notch1在ALS转基因鼠动物模型和细胞模型中表达增多,提示Notch1信号通路可能与ALS相关。展开更多
基金supported in part by an Oversea Study Fellowship from the China Scholarship Council,No.2008630089
文摘Previous studies have confirmed that the beclin 1 complex plays a key role in the initial stage of autophagy and deregulated autophagy might involve in amyotrophic lateral sclerosis. However, the mechanism underlying altered autophagy associated with the beclin 1 complex remains un- clear. In this study, we transfected the Cu/Zn superoxide dismutase 1 G93A mutant protein into the motor neuron-like cell line NSC34 cultured in vitro. Western blotting and co-immunopre- cipitation showed that the Cu/Zn superoxide dismutase 1 G93A mutant enhanced the turnover of autophagic marker microtubule-associated protein light chain 3II （LC3Ⅱ） and stimulated the conversion of EGFP-LC3Ⅰ to EGFP-LC3Ⅱ, but had little influence on the binding capacity of the autophagy modulators ATG14L, rubicon, UVRAG, and hVps34 to beclin 1 during auto- phagosome formation. These results suggest that the amyotrophic lateral sclerosis-linked Cu/Zn superoxide dismutase I G93A mutant can upregulate autophagic activity in NSC34 cells, but that this does not markedly affect beclin 1 complex components.
文摘The expression of cytosolic phospholipase A2 (cPLA2) expression is up-regulated in animal model of ALS and in patients with familial amyotrophic lateral sclerosis (fALS). Inhibition of cyclooxygenase 2 (COX2), which is a downstream enzyme of cPLA2, ameliorates the impairment of motor function in the ALS model mice. Therefore, the arachidonic acid cascade, including the cPLA2-COX2 pathway, is an important therapeutic target of ALS. The current study was designed to investigate the potential of AK106-001616, an inhibitor of cPLA2, in protection of motor neuron cell death induced by mutant superoxide dismutase (SOD1<sup>G93A</sup>). AK106-001616 (1 - 10 μM) protected NSC34 cells (mouse motor neuron like cells) against SOD1<sup>G93A</sup>-induced motor neuron cell death. Furthermore, aspirin, an inhibitor of COX1/2, reduced the SOD1<sup>G93A</sup>-induced motor neuron cell death at a concentration that inhibited COX2. Celecoxib, a selective COX2 inhibitor, also reduced the SOD1<sup>G93A</sup>-induced motor neuron cell death. These results suggest that the arachidonic acid cascade is important for SOD1<sup>G93A</sup>-induced motor neuron cell death and AK106-001616 has a potent neuroprotective effect against it. AK106-001616 may be a useful therapeutic agent against SOD1<sup>G93A</sup>-induced ALS.
文摘目的:观察Notch1在肌萎缩侧索硬化症(ALS)转基因鼠动物模型和细胞模型中的表达情况。方法:应用免疫荧光、免疫印迹、RT-PCR,检测Notch1在95、108、122 d ALS转基因鼠脊髓中的表达变化;检测转染pEGFP-wt-SOD1和pEGFPG93A-SOD1的NSC34细胞模型中Notch1的表达变化。结果:Notch1可与β-tubulinⅢ共表达,与GFAP无明显共表达。较同窝野生型鼠,Notch1于蛋白水平和mRNA水平上的表达在95 d ALS转基因鼠脊髓中无明显变化,在108 d和122 d ALS转基因鼠脊髓中明显升高;与转染pEGFP-wt-SOD的NSC34细胞相比,转染pEGFP-G93A-SOD1的NSC34细胞中Notch1蛋白和mRNA表达增多。结论:Notch1在ALS转基因鼠动物模型和细胞模型中表达增多,提示Notch1信号通路可能与ALS相关。
