【摘要】：目的研究MKP1在Aβ所致MAPK激活、神經(jīng)炎癥和細(xì)胞凋亡中的調(diào)控作用。方法在細(xì)胞培養(yǎng)基中加入不同濃度的Aβ42(0μmol/L,0.1μmol/L,1μmol/L,10μmol/L和100 v),處理24 h后CCK-8檢測細(xì)胞活性。向細(xì)胞培養(yǎng)基中加入10μmol/L的Aβ42,在不同時(shí)間點(diǎn)(0 h、6 h、12 h、18 h和24 h)通過qRT-PCR和Western blot檢測MKP1表達(dá)。將野生型PC12細(xì)胞分為對照組(Control)和Aβ42組,敲除MKP1的細(xì)胞為MKP1 KD+Aβ42組,過表達(dá)MKP1細(xì)胞為MKP1+Aβ組,后3組加入10μmol/L Aβ42,置于培養(yǎng)箱中24 h,通過線粒體膜電位、DCFH-DA以及超氧化物歧化酶活性和丙二醛檢測評價(jià)細(xì)胞內(nèi)氧化應(yīng)激水平,通過qRT-PCR檢測TNF-α和IL-1β表達(dá),Western blot檢測p-JNK水平。結(jié)果發(fā)現(xiàn)經(jīng)Aβ刺激后,PC12細(xì)胞活性受到抑制,MAPK的重要調(diào)節(jié)因子MKP1表達(dá)下調(diào),并呈時(shí)間依賴性。過表達(dá)MKP1后,Aβ誘導(dǎo)的PC12細(xì)胞中p-JNK水平降低,細(xì)胞內(nèi)活性氧類水平下降,TNF-α和IL-1β表達(dá)減少。而敲除MKP1可加重Aβ誘導(dǎo)的PC12氧化應(yīng)激和炎癥反應(yīng)。結(jié)論 Aβ通過下調(diào)MKP1表達(dá)激活MAPK信號途徑,過表達(dá)MKP1可通過抑制JNK信號途徑減輕Aβ所誘導(dǎo)的氧化應(yīng)激和神經(jīng)炎癥從而發(fā)揮神經(jīng)保護(hù)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- 偽 and IL-1 尾 in TNF- 偽 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- 偽 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.
【作者單位】： 華北電力大學(xué)醫(yī)院口腔科;吉林大學(xué)第二醫(yī)院神經(jīng)內(nèi)科;淄博市第一醫(yī)院中西醫(yī)結(jié)合科;吉林大學(xué)第一醫(yī)院神經(jīng)內(nèi)科;延邊大學(xué)附屬醫(yī)院神經(jīng)內(nèi)科;
【分類號】：R749.16

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