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目的观察和探讨在铝致神经细胞死亡中,坏死抑制剂1(Nec-1)对凋亡和自噬的作用。方法用体外原代培养小鼠神经细胞的方法,制造铝损伤神经细胞模型,然后用细胞活力检测、荧光定量PCR(qRT-PCR)、蛋白印迹(Western-Blot)以及流式细胞术等方法从多角度对Nec-1的作用进行研究。结果①流式结果显示,在Al3+(2mmol/L)作用于神经细胞后,随着Nec-1剂量的增加,细胞的凋亡率呈下降趋势,且差异有统计学意义(P<0.05);②qRT-PCR结果显示,以Al3+(2mmol/L)为对照,Nec-1(60和90μmol/L)可使神经细胞的凋亡和自噬相关基因的表达呈显著性下降(P<0.05);③Western-Blot结果显示,同Al3+(2mmol/L)组比,Nec-1(60和90μmol/L)组凋亡相关蛋白caspase-3的表达下降(P<0.01),Nec-1(30和60μmol/L)可使自噬相关蛋白LC3-Ⅱ表达下降(P<0.05)。结论在本试验条件下,Nec-1可减少铝导致的神经细胞凋亡和自噬,起到保护神经细胞的作用。
Objective To observe and explore the effect of necrosis inhibitor 1 (Nec-1) on apoptosis and autophagy in the death of aluminum induced neurons. Methods The primary cultured rat neurons were cultured in vitro to establish a model of injured aluminum cells. The cells were harvested and stained with the method of cell viability assay, qRT-PCR, Western-Blot and flow cytometry A multi-angle study of the role of Nec-1. Results ① The results of flow cytometry showed that the apoptotic rate of Nec-1 cells decreased with the increase of Nec-1 dose after Al3 + (2mmol / L) treatment. The difference was statistically significant (P <0.05). ② The results of qRT-PCR showed that Nec-1 (60 and 90μmol / L) significantly decreased the apoptosis of neurons and the expression of autophagy-related genes (P <0.05) with Al3 + ③Western-Blot results showed that the expression of caspase-3 in Nec-1 (60 and 90μmol / L) group was significantly lower than that in Al3 + (2mmol / L) / L) decreased the expression of LC3-Ⅱ (P <0.05). Conclusion Under the experimental conditions, Nec-1 can reduce the aluminum-induced neuronal apoptosis and autophagy, and play a role in the protection of nerve cells.