Nec-1对铝作用下神经细胞活力及凋亡、自吞噬基因的影响

Effect of Nec-1 on Cell Viability and Genes of Apoptosis and Autophagy in the Aluminum-induced Nerve Cells

[目的]研究坏死性抑制剂Necrostatin-1(Nec-1)对铝(Al)作用下的神经细胞活力及凋亡、自吞噬基因的影响。[方法]体外原代培养小鼠神经细胞,通过2mmol/L Al3+作用于神经细胞制造铝损伤模型,加入不同剂量的Nec-1,用细胞活力检测(CCK-8)和逆转录-聚合酶链反应(RT-PCR)的方法观察Nec-1对染铝神经细胞的作用。[结果]细胞活力检测:以2mmol/L Al3+染毒组作为对照,30μmol/L Nec-1组的细胞活力与对照组比较,差别有统计学意义(P<0.05),60、90μmol/L Nec-1组与对照组比较,差异均有统计学意义(P<0.01)。RT-PCR结果:2mmol/L Al3+染毒组caspase-3基因的表达为0.98±0.120,而Nec-160μmol/L组和Nec-190μmol/L组caspase-3的表达分别为0.50±0.077和0.44±0.050,同对照组相比,两组caspase-3的表达均下降(P<0.01)。2mmol/LAl3+染毒组的LC3-Ⅱ基因的表达为1.82±0.047,而60μmol/L Nec-1组和90μmol/L Nec-1组的LC3-Ⅱ表达分别为1.00±0.022和0.97±0.035,同对照组相比,两组LC3-Ⅱ的表达均呈下降(P<0.01)。[结论]Nec-1可使铝作用下的神经细胞活力上升,并使神经细胞的自吞噬和凋亡基因表达下降。

[ Objective ] To explore the effect of Nee-1 on cell viability and genes of apoptosis and autophagy of the aluminun-induced nerve cells. [ Methods ] We conducted the study by means of cell culture in vitro, producing aluminum injury model by Al3＋（ 2 mmol/L ）, then intervented cell death by Necrostatin- 1 （ Nee-1 ）, and investigated the Nee-1 ＇s role toward the Al- induced nerve cells using CCK-8 and RT-PCR methods. [ Results ] Cell viability of Nee-1 （ 30 μmol/L ）group had statistically difference （ P 〈 0.05 ）, whereas of the Nec-1 （ 60 μmol/L ） and Nee-1 （ 90 μmol/L ） groups had very significant difference （ P 〈 0.01 ）as compared with AP（ 2 mmol/L ）exposure group （ control ）.The RT-PCR demonstrated that the expression of caspase-3 gene in Al3＋ （ 2 mmol/L ） group （ control ） was 0.98 ± 0.120, but that of Nee- 1 （ 60 μmol/L ） and Nee- 1 （ 90 μmol/L ） groups were 0.50 ＋ 01077 and 0.44 ＋ 0.050 respectively. Compared with the control group, the expression of caspase-3 gene of the latter two groups decreased with statistical significance（ P 〈 0.01 ）. The expression of LC3-Ⅱgene of Al3＋（ 2 mmol/L ）group（ control ）was 1.82 ± 0.047, but that of Nee-1 （60 μmol/L ）and Nee-1 （90 μmol/L ）groups were 1.00 ± 0.022 and 0.97 ± 0.035 respectively. Compared with the control group, the expression of LC3-Ⅱ gene of the latter two groups also decreased with statistical significance（ P 〈 0.01 ）. [ Conclusion ] In summary, Nee-1 can increase cell viability of Al-induced nerve cells, and decrease the expression of autophagy and apoptosis genes.