Nec-1对铝作用下神经细胞活力及凋亡、自吞噬基因的影响

徐丽; 张勤丽; 牛侨

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

摘要

摘要:
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 2 mmol/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 2 mmol/L Al³⁺染毒组作为对照, 30 μmol/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、90 μmol/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:2 mmol/L Al³⁺染毒组 caspase-3基因的表达为 0.98& #177;0.120,而 Nec-1 60 μmol/L组和 Nec-1 90 μmol/L组 caspase-3的表达分别为 0.50& #177;0.077和 0.44& #177;0.050, 同对照组相比, 两组 caspase-3的表达均下降(P < 0.01)。2 mmol/L Al³⁺染毒组的 LC3-Ⅱ基因的表达为1.82& #177;0.047,而 60 μmol/L Nec-1组和 90 μmol/L Nec-1组的 LC3-Ⅱ表达分别为 1.00& #177;0.022和 0.97& #177;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。

Abstract:
Objective To explore the effect of Nec-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(Nec-1), and investigated the Nec-1's role toward the Alinduced nerve cells using CCK-8 and RT-PCR methods.

Results Cell viability of Nec-1(30 μmol/L)group had statistically difference(P < 0.05), whereas of the Nec-1(60 μmol/L)and Nec-1(90 μmol/L)groups had very significant difference(P < 0.01)as compared with Al³⁺(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& #177;0.120, but that of Nec-1(60 μmol/L)and Nec-1(90 μmol/L)groups were 0.50& #177;0.077 and 0.44& #177;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-Ⅱ gene of Al³⁺(2 mmol/L)group(control)was 1.82& #177;0.047, but that of Nec-1(60 μmol/L)and Nec-1(90 μmol/L)groups were 1.00& #177;0.022 and 0.97& #177;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, Nec-1 can increase cell viability of Al-induced nerve cells, and decrease the expression of autophagy and apoptosis genes.

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