MKP1通过抑制JNK信号途径减轻淀粉样蛋白的神经毒性作用

发布时间：2018-11-16 17:01

【摘要】：目的研究MKP1在Aβ所致MAPK激活、神经炎症和细胞凋亡中的调控作用。方法在细胞培养基中加入不同浓度的Aβ42(0μmol/L,0.1μmol/L,1μmol/L,10μmol/L和100 v),处理24 h后CCK-8检测细胞活性。向细胞培养基中加入10μmol/L的Aβ42,在不同时间点(0 h、6 h、12 h、18 h和24 h)通过qRT-PCR和Western blot检测MKP1表达。将野生型PC12细胞分为对照组(Control)和Aβ42组,敲除MKP1的细胞为MKP1 KD+Aβ42组,过表达MKP1细胞为MKP1+Aβ组,后3组加入10μmol/L Aβ42,置于培养箱中24 h,通过线粒体膜电位、DCFH-DA以及超氧化物歧化酶活性和丙二醛检测评价细胞内氧化应激水平,通过qRT-PCR检测TNF-α和IL-1β表达,Western blot检测p-JNK水平。结果发现经Aβ刺激后,PC12细胞活性受到抑制,MAPK的重要调节因子MKP1表达下调,并呈时间依赖性。过表达MKP1后,Aβ诱导的PC12细胞中p-JNK水平降低,细胞内活性氧类水平下降,TNF-α和IL-1β表达减少。而敲除MKP1可加重Aβ诱导的PC12氧化应激和炎症反应。结论 Aβ通过下调MKP1表达激活MAPK信号途径,过表达MKP1可通过抑制JNK信号途径减轻Aβ所诱导的氧化应激和神经炎症从而发挥神经保护作用。
[Abstract]:Objective to investigate the role of MKP1 in A 尾 -induced MAPK activation, neuritis and apoptosis. Methods different concentrations of A 尾 42 (0 渭 mol/L,0.1 渭 mol/L,1 渭 mol/L,10 渭 mol/L) and 100 v), were added to the cell culture medium for 24 h to determine the cell viability. 10 渭 mol/L A 尾 42 was added to the cell culture medium. The expression of MKP1 was detected by qRT-PCR and Western blot at different time points (0 h, 6 h, 12 h, 18 h and 24 h). The wild-type PC12 cells were divided into two groups: (Control) and A 尾 42, MKP1 KD A 尾 42 for knockout MKP1, MKP1 A 尾 for MKP1 A 尾, and 10 渭 mol/L A 尾 42 for 10 渭 mol/L A 尾 42 for the last three groups, and then put in incubator for 24 h, and passed through mitochondrial membrane potential. The activity of DCFH-DA, superoxide dismutase (SOD) and malondialdehyde (MDA) were measured to evaluate the level of oxidative stress in cells, and the expression of TNF- 伪 and IL-1 尾 in TNF- 伪 and IL-1 尾 was detected by qRT-PCR to detect the level of p-JNK. The results showed that the activity of PC12 cells was inhibited and the expression of MKP1, an important regulatory factor of MAPK, was down-regulated in a time dependent manner. After overexpression of MKP1, the level of p-JNK and the expression of TNF- 伪 and IL-1 尾 in PC12 cells induced by A 尾 decreased, while the levels of reactive oxygen species in cells decreased. Knockout of MKP1 increased PC12 oxidative stress and inflammatory response induced by A 尾. Conclusion A 尾 activates MAPK signaling pathway by down-regulation of MKP1 expression, and overexpression of MKP1 can relieve oxidative stress and neuroinflammation induced by A 尾 by inhibiting JNK signaling pathway, thus exerting neuroprotective effect.
【作者单位】：华北电力大学医院口腔科;吉林大学第二医院神经内科;淄博市第一医院中西医结合科;吉林大学第一医院神经内科;延边大学附属医院神经内科;
【分类号】：R749.16

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